Proceedings of the Australasian Association of Clinical Biochemists’ 53rd Annual Scientific Conference

. 2015 Aug;36(Suppl):S1–S49.

Proceedings of the Australasian Association of Clinical Biochemists’ 53^rd Annual Scientific Conference

PMCID: PMC4745263

INVITED SPEAKERS	PAGE
S1 HBA1C FOR DIAGNOSIS (INCLUDING PREGNANCY AND IN CHILDREN) A McElduff	S11
S2 IMPLEMENTATION OF HBA1C AS A DIAGNOSTIC TEST IN NEW ZEALAND CM Florkowski	S11
S3 THE INTRODUCTION OF HBA1C AS A DIAGNOSTIC TEST IN AUSTRALIA G Jones	S11
S4 TIME TO RETIRE MICROALBUMIN? – NEGATIVE G Jones	SEE CONFERENCE HANDBOOK
S5 TIME TO RETIRE MICROALBUMIN? - AFFIRMATIVE R MacIssac	S11
S6 BIOCHEMISTRY OF PHAEOCHROMOCYTOMA G Eisenhofer	S12
S7 LABORATORY TESTING FOR PRIMARY ALDOSTERONISM IN 2015 – CURRENT SITUATION AND IMPORTANT NEW DEVELOPMENTS M Stowasser	S12
S8 THE PERFORMANCE/ADEQUACY OF CURRENT LABORATORY ASSAYS FOR GH AND IGF1 KKY Ho	S12
S9 GETTING THE RIGHT ANSWER – THE IMPORTANCE OF TRACEABILITY G Jones	S12
S10 REVIEW OF QAP ACTIVITIES K Barancek	S13
S11 CLOSING PLENARY – HYPOTHETICAL T Badrick	SEE CONFERENCE HANDBOOK
S12 DIABETIC HYPERLIPIDAEMIA D Sullivan	S13
S13 FIBROGENESIS BIOMARKERS IN DIABETES COMPLICATIONS SM Twigg	S13
S14 ADVANCED GLYCATION IN DIABETES AND EFFECT OF DIET J Forbes	S13
S15 INSULIN ASSAYS FIT FOR PURPOSE? P Williams, N Pereria, MS Twigg, T Yen	S13
S16 MECHANISMS OF β-CELL FAILURE IN TYPE 2 DIABETES R Laybutt	S14
S17 CLOSING THE LOOP OR RUNNING IN CIRCLES? MAKING PROGRESS J Gunton	S14
S18 ASSESSMENT OF POLYCYSTIC OVARY SYNDROME A Joham	S14
S19 FGF-21 AS A METABOLIC HORMONE C Sue	SEE CONFERENCE HANDBOOK
S20 BMAL AND THE METABOLIC CLOCK D Kennaway	SEE CONFERENCE HANDBOOK
S21 STEROID ANALYSIS BY LC-MS M Wright	S14
S22 MACROPROLACTIN – CURRENT PROCESSES AND RECOMMENDATIONS L Boscato	S15
S23 ASSAYS FOR MONITORING CUSHING SYNDROME A McCormack	SEE CONFERENCE HANDBOOK
S24 CORTISOL RESPONSES CAN IDENTIFY INDIVIDUALS WITH INCREASED SUSCEPTIBILITY TO OBESITY B Henry	SEE CONFERENCE HANDBOOK
S25 MOLECULAR DIAGNOSTICS IN THYROID CANCER C Woolnough	S15
S26 MOLECULAR TESTING IN PHAEOCHROMOCYTOMA D Benn	S15
S27 METANEPHRINE TESTING - WHY, HOW AND WHEN? G Woollard, M Whiting, for the AACB Biogenic Amines Working Party	S15
S28 QUALITY OF HBA1C MEASUREMENTS IN AUSTRALIA –A REPORT FROM THE RCPAQAP P Williams, S Koetsier, V Braniff, G Jones	S16
S29 ENDOCRINE PROGRAM – A WORKING REVIEW L Boscato	S16
S30 BIOCHEMICAL ASSESSMENT OF “BONE HEALTH” EM Lim	S16
S31 HYPOPHOSPHATAEMIA R J Clifton-Bligh	S16
S32 CALCIUM-SENSING RECEPTORS: MAKING SENSE OF CALCIUM REGULATION AD Conigrave	S17
S33 ENDOCRINE PROGRAM ANALYTE UPDATE L Jolly	SEE CONFERENCE HANDBOOK
S34 COMMON RESULTS: ARE WE GOOD ENOUGH YET? S Koetsier	S17
S35 INTERPRETING EQA AND ASSAYS AT RISK T Badrick	S17
S36 LC-MS VITAMIN D - A GAME CHANGER FOR LABS M Wright	S17
S37 HUMAN HEALTH AND VITAMIN D STATUS; ‘WHY VITAMIN D TRIALS AND META-ANALYSES OFTEN INDICATE NEGATIVE RESULTS’ HA Morris	S17
S38 VITAMIN D: HYPE, HOPE AND REALITY CM Florkowski	S17
S39 QAP CLINICAL CASES K Barancek, G Jones, H Martin, L Nguyen, G Ward	S18
ORAL PRESENTATIONS
A1 DRIED BLOOD SPOT 17 HYDROXYPROGESTERONE MEASUREMENT UTILISING ON-LINE SPE COUPLED TO UPLC-MS/MS BC McWhinney, AJ Wilce, JPJ Ungerer	S19
A2 PLASMA FREE METANEPHRINES BY LC-MSMS: COMPARISON OF METHODS IN USE ACROSS AUSTRALIA AND NEW ZEALAND M Whiting, G Woollard, B McWhinney, A Ellis, K Hoad, T Novos, T Andersen, S Koetsier	S19
A3 EVALUATION OF ABBOTT ARCHITECT URINE FREE CORTISOL CUT-OFF G Rathnayake, I Bretherton, Q Lam, B McWhinney, M Grossmann, J Grant, M Harrop, H Schneider, M Mohr, C Chiang	S19
A4 HORMONE MODELLING IN PRETERM NEONATES: ESTABLISHMENT OF SERUM STEROID REFERENCE INTERVALS BY LC-MS/MS RF Greaves, J Pitkin, CS Ho, J Baglin, RW Hunt, MR Zacharin	S19
B1 ANTIBODY INTERFERENCE IN ACTH IMMUNOASSAY COMPLICATING MANAGEMENT OF CUSHING’S DISEASE KW Choy, J Teng, N Wijeratne, CY Tan, JCG Doery	S20
B2 CORTICOSTEROID-BINDING GLOBULIN CLEAVAGE IS PARADOXICALLY REDUCED IN ALPHA-1 ANTITRYPSIN DEFICIENCY: IMPLICATIONS FOR CORTISOL HOMEOSTASIS MA Nenke, M Holmes, W Rankin, JG Lewis, DJ Torpy	S20
B3 LABORATORY MANAGEMENT OF HAEMOLYSED SAMPLES – RESULTS OF AN RCPAQAP SURVEY P Petinos	S20
B4 SUPPORTING GLUCOSE TESTING OVER 1 MILLION SQUARE KILOMETRES R Tirimacco, P Simpson, P Cowley, L Siew, P Tideman	S21
B5 ANALYSIS OF TROPONIN T DELTA CHANGES FOR THE DIAGNOSIS OF AMI P Tideman, R Tirimacco, P Simpson, P Cowley, L Siew, N Berry	S21
C1 MODIFIED ABBOTT C16000 URINE PROTEIN METHOD FOR PATIENTS WITH DILUTE URINE R Cherrie, R Flatman, C Ison, G Ward, L Price	S21
C2 MEASUREMENT UNCERTAINTY FOR THE CLINICAL LABORATORY – A REVISION OF THE CONCEPT GRD Jones	S22
C3 THE RELATIONSHIP BETWEEN ROUTINE VITAMIN B6 LEVELS AND NON-P5P TRANSAMINASE ASSAY ACTIVITY KA Sikaris, C Trambas, T Yen, D Kanowski, L Price, M Freemantle, ZX Lu	S22
C4 WHAT IS AN EQUIVOCAL SERUM TOTAL VITAMIN B12 CONCENTRATION? ZX Lu, JV Warner, J Chang, JCG Doery, AR McNeil, KA Sikaris	S22
C5 ASSESSING THE QUALITY OF LABORATORY MEASUREMENTS OF HORMONES IN THE 2015 RCPA QAP ENDOCRINE PROGRAM V Vamathevan, J Murby, L Jolly, K Barancek, G Jones	S23
POSTER PRESENTATIONS
P1 URINARY CATECHOLAMINES MEASUREMENT BY LC/MS/MS AC Yeoh, P Staneshwar, BBA Tan	S24
P2 URINARY METANEPHRINE BY LC/MS/MS FOR CLINICAL USE AC Yeoh, P Staneshwar, BB Tan	S24
P3 NEW CUT-OFF VALUE OF THE VIDAS CMV IgG AVIDITY GIVE LESS BORDERLINE RESULTS A Lufiandati, Y Liau	S24
P4 MULTISTAGE FRAGMENTATION OF 1α, 25(OH)2-VITAMIN D3 COMBINED WITH METABOLITE PROFILE BY LCMSMS UTILISING A QUADRUPOLE LINEAR ION TRAP BR Cooke, M Wright, C Hodgkins	S24
P5 STEROID ANALYSIS IN DIFFERENT MATRICES UTILISING STANDARDISED ON-LINE SPE COUPLED TO UPLC-MS/MS BC McWhinney, AJ Wilce, JPJ Ungerer	S25
P6 IMPROVED MEASUREMENT OF PLASMA FREE METANEPHRINES UTILISING ON-LINE SPE COUPLED TO UPLC-MS/MS B Myers, BC McWhinney, JPJ Ungerer	S25
P7 VALIDATION OF A SHORTER SPIN TIME OF BD VACUTAINER PST TUBES FOR URGENT SPECIMENS C Woods, C Oakman, J Farquhar, P Hickman, D Hughes	S25
P8 ROUTINE LIPID PARAMETERS IN THE PREDICTION OF SMALL DENSE LDL KA Sikaris, C Trambas, N Taylor, G Watts, R Shirley, T Partridge, R Grant	S26
P9 EVALUATION OF THE PRECISION AND ACCURACY OF HBA1C ASSAYS C Trambas, KA Sikaris, S Heathcote, R Woodburn, K Parfrey, E Maxwell, ZX Lu	S26
P10 PARAPROTEIN INTERFERENCE IN AUTOMATED CHEMISTRY SDC Thomas, R Lavery, J Wei	S26
P11 RESPINNING LITHIUM HEPARIN TUBES AFTER REFRIGERATION FOR ADD ON REQUESTS E Moore, J Estacio, J Farquhar, P Hickman, D Hughes	S26
P12 i-STAT TROPONIN I – SAMPLE TYPE ANALYSIS AND COMPARISON WITH TROPONIN T FL Groeneveld, M Roser, GRD Jones	S27
P13 BORIC ACID DOES NOT AFFECT THE MEASUREMENT OF URINE CREATININE, SODIUM, POTASSIUM, CALCIUM, CHLORIDE AND PHOSPHATE FB Mohd Abu Bucker, SK Sethi, TP Loh	S27
P14 ANALYTICAL EVALUATION OF THE ROCHE cobas b 101 HbA1c ASSAY G Dimeski, K Bassett, J Johnston, N Brown	S27
P15 EVALUATION OF ROCHE COBAS 6500 URINE ANALYZER IN A DIAGNOSTIC LABORATORY J Jabon, LZ Ong, S Saw, SK Sethi	S27
P16 CASE REPORT: MACRO-TROPONIN T GRD Jones, J Chung	S28
P17 CSF BILIRUBIN MEASUREMENTS ON ROCHE MODULAR – EFFECT OF ASSAY REFORMULATION J Chung, GRD Jones	S28
P18 VALIDATION OF BIOCHEMICAL TESTING OF FLUID SAMPLES BY DILUTION INTO A SERUM MATRIX: A PRELIMINARY INVESTIGATION J Chung, GRD Jones	S28
P19 PERFORMANCE EVALUATION OF C-REACTIVE PROTEIN AND PROCALCITONIN TEST ON THE ichroma^TM Smart ANALYZER J Yoo, JH Rim, HS Lim, JR Choi	S29
P20 A NOVEL APPLICATION OF LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY TECHNOLOGY FOR ESTABLISHING THE PRESENCE OF TOPICAL STEROIDS IN A SKIN WHITENING CREAM K Waller, A Hall, G Marshall, JV Warner	S29
P21 N-ACETYLCYSTEINE INTERFERENCE IN THE ABBOTT AND CAMBRIDGE LIFE SCIENCES PARACETAMOL ASSAYS L Nguyen, L Koe, D Chesher	S29
P22 LOW URINE pH CAN CAUSE FALSE-HIGH MEASUREMENT OF 5-HIAA BY LC-MSMS M Whiting	S29
P23 EVALUATION OF THE EFFECT OF ABNORMAL HAEMATOCRIT LEVELS UPON THE ACCURACY OF HOSPITAL GRADE BLOOD GLUCOSE METERS P Talsma, P Hickman, J Potter	S30
P24 EVALUATION OF CREATININE AND CHLORIDE ON THE EPOC BLOOD GAS AND ELECTROLYTE POINT OF CARE ANALYSER R Tirimacco, PA Simpson, L Siew, P Cowley, N Velleman, PA Tideman	S30
P25 CHANGE IN STATUS OF SUNSHINE VITAMIN WITH GENERAL AWARENESS: STILL LOTS TO BE DONE S Kaur, S Puri, S Parminder, I Verma	S30
P26 REVERSE PHASE COLUMN CHROMATOGRAPHY (RPCC) FOR HIGH SPEED SEPARATION OF AMINO ACIDS AND ITS DERIVATIVES WITHIN BASIC AND LIMITED RESOURCE REDUCED SETTINGS V Lal, A Kumar, M Parakh	S31
P27 WHICH SPECIMEN TUBE IS BEST FOR SERUM/PLASMA OR WHOLE BLOOD TRACE ELEMENT ANALYSIS? R Bahnisch, J Clark, W Rankin, M Saleem	S31
P28 USUAL PATTERN OF SERUM FREE THYROXINE DISTRIBUTION IN CHLDREN ON THYROXINE FOR HYPOTHYROIDISM LK Chin, L Dentrinos, KW Choy, JCG Doery, ZX Lu, P Bergman	S31
P29 METROLOGICAL TRACEABILITY OF ARCHITECT ALKALINE PHOSPHATASE AND AMYLASE ASSAYS TO IFCC REFERENCE METHODS D Armbruster, D Yahalom, L Lennartz, M Orth	S31
P30 ASSESSING ANALYTICAL QUALITY OF Hb A1c ASSAYS USING REFERENCE SAMPLES, ACCURACY BASED GRADING, AND SIGMA METRICS D Armbruster, G Maine	S32
P31 LIPOPROTEIN(a) STATUS AND LDL-CHOLESTEROL IN TYPE 2 DIABETES MELLITUS WITH MICROVASCULAR COMPLICATIONS KA Jhuma, ASM Giasuddin, AMM Haq	S32
P32 OBESITY RELATED METABOLIC ABNORMALITIES IN A GROUP OF SRI LANKAN CHILDREN L Ginige, D Samaranayake, MM Gunatillaka, VP Wickramasinghe	S32
P33 FLAT ORAL GLUCOSE TOLERANCE TEST REVISITED M M Salib, A Simpson, P E Hickman, S Apostoloska, C Yu, J M Potter	S33
P34 BIO-RAD D100 HBA1C – THE SAME CHROMATOGRAPHY IN HALF THE TIME RI King, CR Hawes, CM Florkowski	S33
P35 METFORMIN DELAYS PROGRESSION TO TYPE 2 DIABETES BY 60% – RESULTS OF TEN YEARS OF ROUTINE CLINICAL USE SC Martin	S33
P36 A PATIENT WITH COMPLETELY UNDETECTABLE HEMOGLOBIN A1C REPORTED AS ZERO PERCENT S Das, SK Gupta	S33
P37 HBA1C ACROSS FOUR PLATFORMS. THE ABBOTT ARCHITECT ENZYMATIC METHOD, BIO-RAD VARIANT HPLC WITH THE ALERE AFFINION AND ROCHE B101 POC IMMUNOASSAY METHODS D Hughes, C Harris, L Burgess, M Vowles, J Estacio, PE Hickman, A Simpson, JM Potter	S34
P38 CALCULATED OSMOLALITY – A REFERENCE INTERVAL BASED ON THE AUSSIE NORMAL STUDY D Hughes, J Potter, N Glasgow, J Cavanaugh, G Koerbin, P Hickman	S34
P39 MONITORING OF PATIENT RESULT MEDIANS B Teis, G Ward, R Flatman, L Price, D Kanowski	S34
P40 INCREASING FERRITIN ACROSS NORMAL GGT REFERENCE INTERVALS IN AUSTRALIAN ADULTS – WHAT IS THE TRUE GGT REFERENCE INTERVAL? D Kanowski, R Flatman, L Price	S34
P41 IS THERE A TOTAL B12 LEVEL BELOW WHICH HOLOTRANSCOBALAMIN ADDS NO VALUE? L Price, B Teis, D Kanowski, G Ward, R Flatman, M Freemantle	S35
P42 ASSESSMENT OF THE BECKMAN COULTER ACCESS AMH ASSAY PERFORMANCE AND ESTABLISHMENT OF NORMAL RANGES K Pearson, M Long, J Prasad, S Khouri, Y Wu, M Bonifacio	S35
P43 DEFINITIVE REFERENCE INTERVALS FOR TSH ON THE ABBOTT ARCHITECT ANALYZER PE Hickman, G Koerbin, JM Potter, D Hughes, N West, J Cavanaugh, N Glasgow, D Armbruster	S35
P44 A BHATTACHARYA ANALYSIS OF URINE FREE CORTISOL T Harrower, L Price, G Ward, D Kanowski	S35
P45 COMPARISON OF FREELITETM AND N LATEX SERUM FREE LIGHT CHAIN ASSAYS IN SUBJECTS WITH END STAGE RENAL DISEASE ON HAEMODIALYSIS A Kennard, C Hawley, J Tate, S Klingberg, C Pretorius, C Hutchinson, P Mollee	S36
P46 PREVALENCE OF VITAMIN D DEFICIENCY & HYPOCALCEMIA IN ADULT HEMATOLYMPHOID MALIGNANCY P Parikh, P Chavan, V Bhat, C Naresh, S Salvi, S Waykar, B Pillai	S36
P47 INCIDENCE OF HYPONATREMIA AND SYNDROME OF INAPPROPRIATE ANTI-DIURETIC HORMONE SECRETION (SIADH) IN HAEMATOPOIETIC STEM CELL TRANSPLANTS: A RETROSPECTIVE ANALYSIS P Chavan, V Bhat, P Poladia, A Karmore, M Poojary, A Gokarn, S Ojha, P Parikh, N Khattry	S36
P48 VITAMIN A AND VITAMIN E DEFICIENCY DEVELOPMENT IN INDONESIA POPULATION J Lilianty, MN Rahman, DA Handayani, M Fattah	S37
P49 A CASE OF SEVERE HYPONATRAEMIA SECONDARY TO A LOW SOLUTE DIET AND PRIMARY POLYDIPSIA JD Smith, HG Schneider	S37
P50 DISCORDANT TROPONIN T VS TROPONIN I RESULTS IN PATIENTS WITH SKELETAL MUSCLE DISEASE M Saleem, H Martin	S37
P51 IS POCT TROPONIN SAFE? P Tideman, R Tirimacco, P Simpson, P Cowley, L Siew, N Berry	S37
P52 SUMMARY OF BIOLOGICAL VARIATION STUDIES FOR HIGH SENSITIVITY TROPONIN T PE Stanford, AR Horvath	S38
P53 REPORTING SERUM CARCINOEMBRYONIC ANTIGEN (CEA) VALUES IN RECTAL CANCER PATIENTS RECEIVING ADJUVANT RADIOTHERAPY BEFORE SURGERY: SPOTLIGHT ON THE UTILITY OF PATIENT DERIVED DATA S Chakraborty, S Banerjee, I Mallick, S Sen, P Roy	S38
P54 FAECAL CALPROTECTIN TESTING PROTOCOL FOR DIAGNOSIS OF INFLAMMATORY BOWEL DISEASE – A NEW ZEALAND EXPERIENCE S Hemmady, T Marcinkowski	S38
P55 CHARACTERISTICS OF INDIVIDUALS WITH ELEVATED TROPONIN I ON A HIGH SENSITIVITY ASSAY BUT NORMAL LEVELS ON A CONTEMPORARY ASSAY R Bender, F Njue, S Vasikaran, R Lambert, J Rankin, D Bell	S39
P56 OUTCOME OF SPECIFIC INTERPRETATIVE COMMENTS SUGGESTING SPECIALIST REFERRAL FOR INDIVIDUALS AT HIGH RISK OF FAMILIAL HYPERCHOLESTEROLAEMIA R Bender, G Edwards, J McMahon, AJ Hooper, GF Watts, JR Burnett, DA Bell	S39
P57 IMPACT OF IL-6 GENE ON MALE INFERTILITY -STUDY ON AN INDIAN POPULATIONS INSTEAD OF “ASSOCIATION OF IL-6 GENE WITH MALE INFERTILITY -STUDY ON AN INDIAN POPULATIONS KK Shukla, P Sharma, AA Mahdi, SN Sankhwar	S39
P58 PRO-(IL-18) AND ANTI-(IL-10) PROMOTER GENETIC VARIATIONS AFFECTS SUSCEPTIBILITY AND THEIR SERUM LEVELS IN PROSTATE CARCINOMA S Dwivedi, P Sharma, A Goel, S Khattri, KK Pant	S39
P59 GENE EXPRESSION OF APOPTOTIC PATHWAY IN MALE FERTILITY P Sharma, KK Shukla, S Chambia, SN Sankhwar	S40
P60 DETERMIMATION OF A REFERENCE INTERVAL FOR IGF1 BY DATA MINING USING THE BHATTACHARYA TECHNIQUE D Kanowski, B Teis, M Freemantle, L Price, G Ward	S40
P61 17-HYDROXYPROGESTERONE ASSAYS – TIME FOR HARMONISATION R Tudball, KL Wan, KW Choy, N Wijeratne, J Montalto, C Tran, L Jolly, JCG Doery	S40
P62 A 17-YEAR AUDIT OF TESTING FOR 21-HYDROXYLASE DEFICIENCY KL Wan, Choy KW, N Wijeratne, JCG Doery	S41
P63 EVALUATION OF ROCHE AND BECKMAN COULTER AUTOMATED ANTI-MULLERIAN HORMONE ASSAYS M Freemantle, B Teis, K Young, G Ward, D Kanowski, L Price	S41
P64 DEVELOPMENT OF A METHOD FOR THE SIMULTANEOUS MEASUREMENT OF SERUM TESTOSTERONE, 17-HYDROXY PROGESTERONE AND ANDROSTENEDIONE BY LIQUID CHROMATOGRAPY – TANDEM MASS SPECTROMETRY (LC-MS/MS) M Henman, K Waller, J Warner	S41
P65 REVIEW OF SEX HORMONE TEST REQUESTS IN A TERTIARY HOSPITAL L Ong, S Saw, SK Sethi	S41
P66 PROLACTIN AND REPRODUCTIVE HORMONE STATUS IN OLIGOMENORRHEIC AND INFERTILE FEMALES R Suwal, AK Nepal, B Gelal, S Gautam, BKL Das, M Lamsal, S Majhi, N Baral	S42
P67 DEVELOPMENT OF AN URINE CORTISOL ASSAY BY LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY R Sahertian, M Wright, P Stanford, C White	S42
P68 INTEGRATING LC-MS/MS INTO A CLINICAL LABORATORY: HANDS-FREE DATA TRANSFER FROM SAMPLE HANDLING TO RESULT ENTRY R Sahertian, M Wright, B Patterson, C Hodgkins, J Teal, C White	S42
P69 A PRAGMATIC APPROACH TO PAEDIATRIC REFERENCE INTERVALS FOR CEREBROSPINAL FLUID (CSF) TOTAL PROTEIN, GLUCOSE AND WHITE BLOOD CELL COUNT USING PUBLISHED EVIDENCE, CLINICAL CONSENSUS AND IN-HOUSE VERIFICATION CKM Ho, NWS Tee	S43
P70 TRIMESTER SPECIFIC REFERENCE INTERVALS FOR THYROID HORMONES IN THE SINGAPORE POPULATION ETH Tan, CKM Ho, GSH Yeo, KYC Kwek, BSM Chern, KH Tan	SEE CONFERENCE HANDBOOK
P71 THYROID FUNCTION TEST REFERENCE INTERVALS DURING PREGNANCY USING THE VITROS 5600 E Haworth, C Chang, T Yen, A Chiriano, S Matthews	S43
P72 TOWARDS AGE-RELATED REFERENCE INTERVALS FOR THE HCG STIMULATION TEST IN CHILDREN WITH SUSPECTED DISORDERS OF ANDROGENISATION WK Choong, KW Choy, J Brown, P Bergman, JCG Doery	S43
P73 PAEDIATRIC REFERENCE INTERVALS FOR IGF1 AND IGFBP3 IN SHORT BUT GROWTH HORMONE-REPLETE CHILDREN JD Newman, KW Choy, TP Loh, JCG Doery	S44
P74 URINE PROTEIN CREATININE RATIO IN PREGNANCY – PERFORMANCE OF BENZATHONIUM CHLORIDE AND ENZYMATIC CREATININE P Ward, S Bandodkar, T Yen	S44
P75 EVALUATION OF NEWBORN SCREENING FOR THE DIAGNOSIS OF CARNITINE UPTAKE DEFICIENCY IN INDIAN POPULATION S Mohapatra, AC Poornima, R Devi, R Cariappa	S44
P76 IMPLEMENTATION OF PLACENTAL GROWTH FACTOR INTO FIRST TRIMESTER SCREENING FOR DOWN SYNDROME S Khouri, M Long, M Bonifacio	S44
P77 HIGH FREQUENCY OF UNDETECTABLE URINE PROTEIN DURING SCREENING FOR GESTATIONAL PROTEINURIA – TRUE LOW PROTEIN OR CLNICAL PROTEINURIA MASKED BY EXCESSIVELY DILUTE URINE? T Yen, E Haworth, A Chiriano, S Matthews	S45
P78 IN VITRO FERTLISATION (IVF) AND FIRST TRIMESTER SCREENING (FTS) FOR DOWN SYNDROME. R Sinnadurai, JM Morris, V Tasevski	S45
P79 A TRAINING AND COMPETENCY ASSESSMENT PROGRAM AT RCPAQAP P Petinos, F Intan	S45
P80 SHOULD WE TARGET SET OUR RCPAQAP MATERIAL WHEN THERE ARE GAPS IN THE TRACEABILITY CHAIN? TARGET ASSIGNMENT OF 17-OHP, ANDROSTENEDIONE, DHEA AND DHT BY GC-MS/MS RF Greaves, L Jolly, MF Hartmann, SA Wudy	S46
P81 ESTABLISHMENT OF AN EXTERNAL QUALITY ASSURANCE PROGRAM FOR SERUM DIHYDROTESTOSTERONE L Jolly, SA Wudy, MF Hartmann, CS Ho, RKT Kam, J Joseph, C Boyder, RF Greaves	S46
P82 UNDERSTANDING QC, EQA AND METHOD EVALUATION - A PROGRESSIVE APPROACH TO PREPARING WORK-READY GRADUATES FN Breen, SL Weier	S46
P83 A SIMPLE TOOL TO IDENTIFY AT RISK ASSAYS USING EQA DATA M Mackay, G Hegedus, TC Badrick	S47
P84 END OF AN ERA – DECOMISSIONING THE ABBOTT TDxFLx FOLLOWING THE ASSESSMENT OF TWO CHEMILUMINESCENT METHOTREXATE IMMUNOASSAYS A Chiriano, C Tran, E Haworth, S Matthews	S47
P85 HOW LOW CAN YOU GO? AN EVALUATION OF THE NEW ABBOTT ARCHITECT ci2000 IMMUNOASSAY IN THE THERAPEUTIC DRUG MONITORING OF METHOTREXATE HM Pawson	S47
P86 RAPID AND EFFICIENT ENZYMATIC HYDROLYSIS OF CODEINE AND MORPHINE GLUCURONIDES IN URINE K Ellis, W McConnell, M Huynh, S Sacks, J Grasko, L Dusci	S47
P87 AN EVALUATION OF THE ABBOTT ARCHITECT ASSAY AGAINST THE ABBOTT TDX METHOTREXATE ASSAY M Al-Hinti, P Williams, E Lin	S48
P88 LC-MSMS ALLOWS DETECTION AND MONITORING OF ANTI-PSYCHOTIC POLYPHARMACY MJ Whiting	S48
P89 LABORATORY EVALUATION OF FOUR DRUGS OF ABUSE IMMUNOASSAY KITS JW Lo, CA Quinlan, W McConnell, S Sacks, J Grasko	S48

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CBR. 2015 Aug;36(Suppl):S11.

S1: HBA1C for Diagnosis (Including Pregnancy and in Children)

A McElduff ¹

Abstract

Glycated haemoglobin reflects the average plasma glucose concentration over the prior 8–12 weeks. It quantifies the risk of macrovascular disease and of the microvascular complications of diabetes mellitus. A number of medical (and technical) issues can confound these relationships. The World Health Organization (WHO) conducted a systematic review of the use of HbA1c as a diagnostic test for diabetes mellitus. An HbA1c of 6.5% was recommended as the cut point for diagnosing diabetes. A value <6.5% does not exclude diabetes diagnosed using glucose tests. The WHO expert group concluded that there was currently insufficient evidence to make any formal recommendation on the interpretation of HbA1c levels below 6.5%.

The WHO now recognises two levels of hyperglycaemia first detected in pregnancy: diabetes mellitus in pregnancy and gestational diabetes. In late pregnancy, for minor degrees of hyperglycaemia (gestational diabetes), glycated haemoglobin has been shown to be less effective in predicting adverse pregnancy outcomes than the traditional 75 g OGTT (Diabetes Care 2012;35:574). However, a HbA1c >6.5% would still indicate diabetes mellitus in pregnancy with the same caveat about values <6.5% outside of pregnancy. Work from NZ suggests an early pregnancy HbA1c ≥5.9% is optimal for detecting diabetes and identifies women at increased risk of adverse pregnancy outcomes (Diabetes Care 2014;37:2953).

There are almost no published data on this issue in children. In NSW, type 2 diabetes is about 10% of diabetes in adolescence. Type 2 diabetes is rare before puberty. Most diabetes in children is diagnosed clinically on symptoms and an appropriately elevated plasma glucose level. The international paediatric guidelines suggests that glycated haemoglobin may be useful as a diagnostic aid in borderline situations or in incidentally discovered mild hyperglycaemia in an asymptomatic child. The test is not recommended for routine use (Pediatric Diabetes 2014;15(Suppl. 20): 4–17).

CBR. 2015 Aug;36(Suppl):S12.

S9: Getting the Right Answer – The Importance of Traceability

G Jones ¹

Abstract

Results of laboratory tests are used for medical decision-making with the aim of improving patient health. A fundamental part of the role of the routine laboratory is getting the right result for a laboratory test. This means having assay with good precision, low bias and freedom from interferences. Bias in particular can affect all results for a test leading to incorrect decisions on some patients. Assay results are derived by comparing values in patient samples with values in the assay calibrators. The values for concentrations in calibrators are set by comparison with other calibrators. As we compare laboratory results with decision points and information in the medical literature from all parts of the world, ideally we need to ensure that all results for a test are comparable across the globe. One part of this is selecting the international reference materials and methods that manufacturers should use for setting calibrator values. The Joint Committee for Traceability in Laboratory Medicine (JCTLM) is a joining of international organisations in measurement and laboratory medicine to help with this task. Manufacturers should use the best available materials and methods to assign values to calibrators and laboratories should select traceable methods and verify accuracy to ensure the best outcome for patients.

CBR. 2015 Aug;36(Suppl):S13.

S10: Review of QAP Activities

K Barancek ^1,^✉

Abstract

The RCPAQAP Chemical Pathology Program is a worldwide leader in the provision of external quality assurance programs offering 49 individual programs to over 2300 participants in 40 different countries. The RCPAQAP Chemical Pathology has seen significant changes over the last few years, most notably in the delivery of electronic reports to participants and the introduction of the myQAP customer portal. Our goal is to make your experience with the RCPAQAP even more efficient and convenient. This Review of Activities will focus on program changes and activities over the last 12 months and any new program initiatives planned for 2016. This session will also look at how the introduction of the myQAP has changed the way participants interact with the Chemical Pathology QAP.

CBR. 2015 Aug;36(Suppl):S13.

S12: Diabetic Hyperlipidaemia

D Sullivan ¹

Abstract

Diabetes is a maladaptive alteration in macronutrient metabolism. It affects not only glucose and carbohydrate metabolism, but also the other major class of macronutrient class, namely lipids. Lipid abnormalities manifest as disturbances of the levels of the lipoproteins that transport lipids in the bloodstream. These disturbances may contribute towards the macrovascular complications of diabetes by influencing the processes that underlie atherosclerosis and thrombosis. Increased energy intake and reduction in activity in affluent and developing societies has resulted in an increase in the prevalence of type 2 diabetes.

Clinical evaluation of lipoprotein metabolism in diabetes involves the measurement of total cholesterol, HDL-C and TG following a 12-hour fast. LDL-C is calculated by the Friedewald equation, but this calculation becomes less reliable as TG levels increase beyond 4 mmol/L. Non-fasting samples have been shown to be a more sensitive marker for the detection of individuals with increased risk of CVD, but the unstandardised nature of non-fasting samples makes them unsuitable for the serial monitoring of lipid status in diabetes. Variations in LDL particle composition complicate the relationship between LDL-C and CVD risk, leading to growing enthusiasm for other risk markers such as NHDL-C or apolipoprotein B, particularly in the presence of elevated levels of TG.

The fundamental differences in the pathophysiology and treatment of type 1 and type 2 diabetes are also manifest in their accompanying changes in lipoprotein metabolism and its contribution towards atherosclerosis and macrovascular disease. Fully treated type 1 diabetes usually causes minimal disturbance in the lipoprotein profile, but the atherogenicity of the diabetic state in type 1, combined with the early age of onset, results in an increased life-long risk of CAD that demands efforts to maintain lipoproteins at target levels. Type 2 diabetes typically causes mild hypertriglyceridaemia with low HDL and small, dense LDL. If triglyceride removal becomes saturated, this picture may rapidly progress to massive hypertriglyceridaemia with associated risk of pancreatitis.

CBR. 2015 Aug;36(Suppl):S13.

S13: Fibrogenesis Biomarkers in Diabetes Complications

SM Twigg ¹

CBR. 2015 Aug;36(Suppl):S14.

S18 Assessment of Polycystic Ovary Syndrome

A Joham ¹

Abstract

PCOS is one of the most common conditions in Australian women, affecting a striking 12 to 21% of reproductive aged women depending on the study population and diagnostic criteria applied. PCOS has significant and diverse clinical implications including reproductive (hyperandrogenism, hirsutism, oligo/anovulation, subfertility), metabolic (metabolic syndrome, insulin resistance, impaired glucose tolerance (IGT), type 2 diabetes mellitus (T2DM), adverse cardiovascular risk profiles and possible increased cardiovascular disease) and psychological features (increased anxiety, depression, body image issues and worsened quality of life). PCOS is essentially a hormonal disorder underpinned by insulin resistance and hyperandrogenism. Obesity is common in PCOS and increases the prevalence and severity of the condition by further exacerbating IR.

PCOS is a heterogeneous condition and there is no single test to diagnose PCOS. Changing definitions and a variety of symptoms have made the path to diagnosis for many women difficult. It is estimated that up to 70% of women with PCOS in the community with PCOS remain undiagnosed. Diagnosis of PCOS is now recommended using the ‘Rotterdam’ criteria and is based on the presence of two of the following three criteria: hyperandrogenism (clinical or biochemical), oligomenorrhoea/amenorrhoea or polycystic ovaries on ultrasound, with exclusion of related reproductive disorders. Insulin resistance is common in PCOS, but is not required for diagnosis, in part because of the lack of accurate methods to measure insulin resistance. Anti Mullerian Hormone (AMH) levels may be elevated in women with PCOS, but this is not currently included in the diagnostic criteria either. PCOS is increasingly being recognised as a metabolic disorder and women with PCOS should undergo regular screening for metabolic complications including impaired glucose tolerance and type 2 diabetes.

CBR. 2015 Aug;36(Suppl):S14.

S21: Steroid Analysis by LC-MS

M Wright ¹

Abstract

The introduction and expansion of automated immunoassays in clinical chemistry and endocrinology had the advantage of increasing the number and diversity of assays and the speed at which they could be performed and results released. Economies of scale and structural efficiencies created the opportunity to incorporate and commercialise the sector through advances in automation and IT. Without trying to sacrifice analytical rigor the business model has shifted toward high throughput non-chromatographed and unseparated specimen analyses at reduced cost. However immunometric analysis of steroids have always had issues around sensitivity, and more importantly specificity, which can leave the clinician with the un-enviable question of “what is the true value of this test result?”. Liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) has the potential to provide a solution to this problem but there are many additional considerations to be taken into account before moving to this technology. This presentation will briefly cover immunoassay and LC-MS/MS techniques, why immunoassays struggle to measure steroids & how regulatory bodies may be a part of the problem rather than the cure, when LC-MS/MS can provide the answer but also the difficulties in knowing what to do with the LC-MS/MS result and situations where the correct answer isn’t always what we had hoped for.

CBR. 2015 Aug;36(Suppl):S14–S15.

S22: Macroprolactin – Current Processes And Recommendations

L Boscato ^1,^✉

Abstract

Macroprolactin, a high molecular weight form of prolactin generally composed of prolactin bound to IgG, was first described over thirty years ago. Although it is biologically inactive macroprolactin is detected by most if not all immunoassays and can lead to falsely increased prolactin levels. Its presence has resulted in misdiagnosis, inappropriate treatment and unnecessary expensive investigation. In contrast to heterophilic antibodies where interference can be easily reduced by the assay manufacturer, macroprolactin is an ongoing problem. It is the role of the laboratory to provide a clinically meaningful prolactin level in the presence of macroprolactin.

Most laboratories now screen samples with a high prolactin level for the presence of macroprolactin using the polyethyleneglycol (PEG) procedure. PEG precipitates immunoglobulin including macroprolactin. A low recovery of prolactin in the supernatant (free, bioactive or monomer prolactin) indicates that most prolactin is in the macroprolactin form and that the prolactin level is falsely elevated. Traditionally laboratories have reported only the pre-PEG prolactin level and presence or absence of macroprolactin generally with a comment that the prolactin was elevated due to macroprolactin if appropriate. As true hyperprolactinaemia can still exist in addition to macroprolactinaemia it is now recommended that the free or monomer prolactin level should be reported with appropriate reference intervals (eg determined using PEG treated specimens).

While reporting the free prolactin level provides a clinical meaningful measurement of prolactin a recent survey of Australian and New Zealand laboratories indicated that there is considerable variability in every step of the procedure, from what constitutes a high prolactin level to the details of PEG procedure and results reporting. There is certainly a need for standardisation of the procedure including reporting. Introduction of a macroprolactin external quality assurance program should assist in the harmonisation of results.

CBR. 2015 Aug;36(Suppl):S15.

S25: Molecular Diagnostics in Thyroid Cancer

C Woolnough ¹

Abstract

Thyroid cancer is diagnosed by cytology of fine needle aspirates (FNA) from thyroid nodules. The cytology results are then classified using the Bethesda scale I–VI, where Bethesda II is benign and Bethesda VI is malignant. The Bethesda III–V categories are classified as undetermined significance to suspicious for malignancy and it is in these intermediate classifications where molecular diagnostic tools could be of the most benefit.

It is well recognised that the different types and variants of thyroid cancer are associated with particular DNA mutations. The current molecular analysis uses samples attained through FNA and formalin fixed paraffin embedded (FFPE) tissue from surgery. FNA sample analysis has greater clinical potential than FFPE, as results from FNA testing guide surgical decisions. Diagnostic methods range from detection of simple DNA point mutations to comprehensive panel testing which detect a range of DNA, RNA and miRNA changes.

The BRAF V600E mutation occurs in 60–80% of patients with papillary thyroid cancer and is typically present in nodules with Bethesda categories of V (suspicious for malignancy) and VI (malignant). It is rarely present in indeterminate Bethesda categories III–IV. BRAF testing of FNA samples may have clinical utility and guide clinical decision making. Another emerging area for molecular diagnostics in thyroid cancer is in the measurement of miRNAs in the tissue and in the circulation. These small non-coding RNAs are critical for a variety of cellular processes and whilst their exact roles are not well understood they can be detected in the blood and as such have potential for non invasive screening for thyroid cancer presence and or progression.

The development of sensitive molecular diagnostic tools for analysis of FNA samples and the potential utility of serum miRNAs as biomarkers for thyroid cancer presence have the capacity to advance thyroid cancer diagnosis.

CBR. 2015 Aug;36(Suppl):S15.

S26: Molecular Testing in Phaeochromocytoma

D Benn ^1,^✉

Abstract

Introduction

This year marks the 15th anniversary of a major discovery by Baysal (USA) which changed the genetic landscape of two rare tumour types, inextricably linking them and three other tumour types. Prior to 2000, 10% of phaeochromocytoma (PC), were associated with the familial cancer syndromes MEN2, VHL or MEN1. Similarly paragangliomas (PGL) of the parasympathetic system were known to occur in families but the genetic basis remained elusive despite extensive linkage studies. Baysal’s discovery in 2000 identified the gene encoding succinate dehydrogenase subunit D (SDHD) as responsible for familial head and neck PGL, and subsequent studies linked these tumours & inherited PC not only to SDHD but also to the other members of mitochondrial complex II, namely SDHC, SDHD and SDHA.

Methods

In 2000 we commenced genetic testing in PC/PGL patients initially for germline mutations in RET, VHL, SDHD, SDHB, SDHC and subsequently SDHA, SDHAF2, TMEM127, MAX and EPAS1. The identification of gastrointestinal stromal tumours, renal cell carcinoma and pituitary tumours with specific pathologies as being SDH deficient, has also expanded the fields of study.

Results

Since 2000 we have tested 604 PC/PGL cases and discovered germline mutation in 163 patients. Together with results from predictive testing in relatives of these probands we have identified 509 individuals with a germline mutation in a PC/PGL gene. To further characterise the phenotype and genotype of these tumours in the Australian population we established the Australian SDH Consortium.

Conclusion

The contribution of genetics to the clinical management of patients and their families with these tumours is a remarkable journey, leading to these tumour syndromes now being recognised as the most highly heritable cancers and exemplifying once again how rare cancers can open a Pandora’s box of discovery.

CBR. 2015 Aug;36(Suppl):S15.

S27: Metanephrine Testing - Why, How and When?

G Woollard ¹, M Whiting ¹; The AACB Biogenic Amines Working Party¹

Abstract

Various biogenic amines and their metabolites are measured for the biochemical diagnosis, and the monitoring of treatment, of neuroendocrine tumours. The AACB Biogenic Amines Working Party (BAWP) continues to monitor the analytical performance of these tests through the relevant EQA programs offered by the RCPAQAP Chemical Pathology group.

The recent publication by the Endocrine Society of clinical practice guidelines for the investigation of phaeochromocytoma and paraganglioma has confirmed the importance of testing for metanephrines (also called metadrenalines), rather than catecholamines. A metanephrine profile (normetanephrine, metanephrine and 3-methoxytyramine) can be obtained either from a 24-hour urine collection, or from plasma collected under appropriate conditions. At present, urine testing involves an acid hydrolysis step to measure the sum of free and conjugated metanephrines, whereas plasma metanephrines are measured as free compounds.

The BAWP has noted the rise of LC-MSMS as an important analytical technique. The proportion of laboratories enrolled at RCPAQAP in 2015 and using LC-MSMS is 10% for urine free catecholamines, 36% for urine total metanephrines and 85% for plasma free metanephrines. Nevertheless, measurement conditions are not standardised, as illustrated in a survey of laboratories performing plasma metanephrine testing across Australasia. The RCPAQAP EQA program for plasma metanephrines has continued to grow internationally with many overseas enrolments. This reflects both the increased uptake of this test by laboratories and the limited number of suppliers of an EQA program for these analytes.

Future directions predicted for metanephrines, which should assist laboratories, are the use of free, rather than total, urine metanephrine measurements, and the possibility of spot, rather than 24-hour, urine collections. Before this can be achieved, appropriate reference ranges need to be derived from large clinical studies to establish diagnostic test sensitivity and specificity for neuroendocrine tumours.

CBR. 2015 Aug;36(Suppl):S15–S16.

S28: Quality of HBA1C Measurements in Australia – A Report From the RCPAQAP

P Williams ¹, S Koetsier ², V Braniff ³, G Jones ⁴, on behalf of the RCPAQAP HbA1c Working Group

Abstract

Introduction

S37: Human Health and Vitamin D Status; ‘Why Vitamin D Trials and Meta-Analyses Often Indicate Negative Results’

HA Morris ¹

Abstract

The well characterised endocrine pathway of vitamin D metabolism and its activities are responsible for vitamin D regulation of plasma calcium and phosphate homeostasis under control of serum 1,25-dihydroxyvitamin D, the biologically active metabolite of vitamin D. This pathway protects against the metabolic bone disease of osteomalacia in adults or rickets in children. The critical level for serum 25-hydroxyvitamin D to maintain adequate serum 1,25-dihydroxyvitamin D is 20 nmol/L. In contrast a large body of data demonstrate associations between an adequate vitamin D status and incidence of osteoporosis as well as many other diseases. The strongest evidence is for osteoporosis, which extends to the relationship between serum 25-hydroxyvitamin D and bone mineral density and reduction of fracture risk. Bone cells metabolise 25-hydroxyvitamin D to 1,25-dihydroxyvitamin D to elicit biological responses including osteoblast maturation, reducing bone resorption, and enhancing mineral retention in bone to reduce the risk of fracture in the elderly. The critical level for serum 25-hydroxyvitamin D for optimising the health of the skeleton is approximately 60 nmol/L. Vitamin D is a threshold nutrient such that an adequate level is required and excess nutrient is catabolised. In osteoporosis there is a critical interaction between vitamin D metabolism and dietary calcium intake. Without an understanding of the physiological relationship between these nutrients any beneficial outcomes on bone tissue can be obscured. Consequently badly designed randomised clinical trials or meta-analyses of such data will produce an incorrect conclusion. The highest quality clinical investigations must be a test of the underlying physiology in the clinical context. Ignorance of the threshold nature of vitamin D and calcium as nutrients, as well as the critical relationship between these nutrients has been the basis for numerous ‘negative’ outcomes of trials of vitamin D supplements.

CBR. 2015 Aug;36(Suppl):S17–S18.

S38: Vitamin D: Hype, Hope and Reality

CM Florkowski ¹

Abstract

Vitamin D has been traditionally linked to bone health, with deficiency causing rickets and osteomalacia. Increasingly, however it has become realised that vitamin D controls the activity of over 200 genes and it has become implicated in a wide range of disease processes, including diabetes, cardio-vascular disease, malignancy and infectious diseases. Coupled with this, is the debate over what constitutes a health plasma 25(OH) vitamin D concentration and whether an optimal level is 50 nmol/L, 70 nmol/L or whatever. If higher concentrations are accepted as healthy, then that renders a significant proportion of our populations as deficient, certainly during winter months and brings into discussion, ways of raising vitamin D, including sunlight exposure (which may increase risk of skin malignancies) and taking dietary supplements. A lot of the evidence linking vitamin D to several conditions is based on case-control studies, which are subject to biases from many confounding factors and reverse-causality, not to mention publication bias. Ideally, a role for vitamin D in any condition should be supported by high quality randomised controlled trials (RCTs). Often, however these are under-powered, of too short duration or use inadequate doses.

A paradigm is in infectious diseases where a role for vitamin D has been advocated on the basis of mostly case-control studies and limited RCTs. Our group undertook an RCT of 322 healthy adults, participants, randomly assigned to initial dose of 200 000 IU then 100 000 IU monthly or placebo for 18 months. There were no statistically significant differences in the number of upper respiratory infections, duration of symptoms or days off work between the two groups. These findings remained unchanged when the analysis was repeated by season and by baseline 25-OHD levels.

The emergence of high quality RCTs is putting the role of vitamin D into better perspective.

(JAMA 2012;308:1333–9)

CBR. 2015 Aug;36(Suppl):S18.

S39: QAP Clinical Cases

K Barancek ^1,^✉, G Jones ¹, H Martin ¹, L Nguyen ¹, G Ward ¹

Abstract

Interpretative commenting is an important aspect of the post-analytical phase in chemical pathology. The introduction of new and complex tests together with increased electronic data communication, use of expert systems, the ability to build interpretative algorithms into laboratory systems and the introduction of clinical and regulatory guidelines are some of the factors that enhance the importance of adding comments to chemical pathology reports.

Commenting on reports is a skill that needs to be acquired during training and continually refined. The RCPAQAP Chemical Pathology Patient Report Comments Program is an educational self-assessment program aimed at the continuing professional development of individuals who currently add comments to reports generated by their laboratories, and the education of individuals who are training for such a role.

Several cases which have previously been distributed in this program will be presented in the QAP Clinical Cases Symposium and discussed with active audience participation. Dissenting views will be encouraged and an interactive format will be followed.

CBR. 2015 Aug;36(Suppl):S19.

A1: Dried Blood Spot 17 Hydroxyprogesterone Measurement Utilising on-Line SPE Coupled to UPLC-MS/MS

BC McWhinney ^1,^✉, AJ Wilce ¹, JPJ Ungerer ¹

Abstract

Introduction

Current mass spectrometry methods for Dried Blood Spot (DBS) 17 Hydroxyprogesterone (17OHP) measurement involve labour intensive liquid-liquid extraction (LLE), dry down, reconstitution and long run times. In this study, a method for measuring 17OHP in DBS has been developed that takes advantage of the capabilities of an online SPE system (ACQUITY UPLC Online SPE Manager) to enable the accurate measurement of 17OHP and overcomes the problems with associated with LLE.

Methods

DBS samples were mixed with deuterated internal standard in a Zinc Sulfate/Methanol solution in 2 mL 96 well plates. After centrifugation, an aliquot of sample was injected into the online SPE equipped UPLC-MS/MS system. Samples underwent an automated Solid Phase Extraction (SPE) utilising a Waters Online Sample Manager, coupled to a UPLC reverse phase chromatographic system. Quantification of 17OHP was achieved by monitoring two transitions on a Waters Xevo TQD mass spectrometer. The assay run time was 4.5 minutes.

Results

The analytical range of the assay was 500 nmol/L. The Limit of the Blank (LOB) was 0.02 nmol/L and the Limit of Detection (LOQ) with a CV of 20% was 0.20 nmol/L. The inter-run imprecision (CV) for quality control samples at 2.5, 25 and 60 nmol/L was less than 6%. Correlation with the CDC newborn screening quality assurance program based upon 30 samples: UPLC-MS/MS = 0.36 + 1.03 CDC (r = 0.997).

In this study, absolute delta change was clearly superior when compared to relative delta change for the diagnosis of AMI. Delta changes should be used in an appropriate way as part of a clinical pathway for the diagnosis of AMI and prevent unnecessary hospital admissions.

CBR. 2015 Aug;36(Suppl):S21–S22.

C1: Modified Abbott C16000 Urine Protein Method for Patients with Dilute Urine

R Cherrie ^1,^✉, R Flatman ¹, C Ison ¹, G Ward ¹, L Price ¹

Abstract

Introduction

Analysis of urine protein/creatinine ratios is an increasingly used parameter in lieu of 24hr urine protein collection, particularly in the diagnosis of pregnancy associated proteinuria. In patients with a combination of low urine protein concentration and dilute urine (i.e. low creatinine concentration), the final calculated ratio may need to be reported as “less than” a value which is higher than the clinical cutoff (e.g. <35 mg/mmol, when values >30 mg/mmol suggest increased pregnancy risk). This situation can be particularly confusing for clinicians.

We noted during a change of method from Roche Integra to Abbott Architect a loss of sensitivity at low concentrations that would increase the likelihood of this scenario (Roche 0.04 g/L, Abbott 0.068 g/L). In our patient population as many as 40% of our urine protein requests have a concentration reported (from Roche Integra) of <0.07 g/L.

Method

In response, Abbott Australia developed a modified urine protein method with increased (3x) sample/reagent ratio, specifically for dilute urine estimations. Only samples with values measured <0.068 g/L were to be re-tested by the modified methodology.

Results

Linear regression by Passing-Bablok between Architect (original method) and Integra across a range of concentrations (0.04 to 10.0 g/L) showed excellent agreement (Slope 1.157, intercept – 0.030, n=30). Within run imprecision (n=20) value on 2 dilute patient samples were as follows:

Instrument	Precision @ 0.04 g/L	Precision @ 0.08 g/L
Integra	23.6%	7.6%
Architect (original)	(below reporting limit)	6.8%
Architect (modified)	3.2%	(not run)

Introduction

Phaeochromocytoma is an uncommon but important cause of hypertension. Its secretory products, catecholamines and their metabolic products, are powerful pressor agents, and severe hypertension and stroke may be the sequelae if the condition is not identified and treated. In this study, a method for simultaneously measuring three biogenic amines (metanephrine, normetanephrine, 3-methoxytryamine) from plasma has been developed that takes advantage of the capabilities of an online SPE system (ACQUITY UPLC Online SPE Manager). This enables the accurate and efficient measurement of these analytes.

Methods

Plasma free metanephrines were measured by an automated on-line Solid Phase Extraction (SPE) coupled to a UPLC HILIC chromatographic system. Plasma samples were diluted 1:1 with deuterated internal standards in a Zinc Sulfate/Acetonitrile solution in 2 mL 96 well plates. After centrifugation, an aliquot of sample was injected into the online SPE equipped UPLC-MS/MS system. Quantification was achieved by monitoring two transitions for each analyte on a Waters Xevo TQD mass spectrometer. The assay time between injections was 4.5 minutes.

Results

The analytical range of the assay was up to 50,000 pmol/L. The Limit of quantitation (LOQ) with a CV of 20% was 20 pmol/L for all 3 analytes. The relationship with our current off-line SPE based upon 250 patients was excellent (r² > 0.97). The inter-run imprecision across 3 levels for the 3 analytes were all < 8%.

Conclusion

Conclusion

Metabolic abnormalities are present significantly in obese children compared to their non-obese counterparts from a younger age. Liver transaminases and hs-CRP were elevated in a significant number of obese children.

CBR. 2015 Aug;36(Suppl):S33.

P33: Flat Oral Glucose Tolerance Test Revisited

MM Salib ^1,^✉, A Simpson ¹, PE Hickman ^1,², S Apostoloska ¹, C Yu ¹, JM Potter ^1,²

Abstract

Aims

This study aimed to determine the demographics and clinical characteristics of individuals with elevated troponin I by a high sensitivity troponin I (hs-TnI) assay, but with non-elevated troponin I (TnI-II) on a contemporary assay. The hs-TnI cut-points for males were ≥26 ng/L and for females ≥16 ng/L.

Methods

Individuals with elevated hs-TnI and non-elevated TnI-II between 18/11/2012 and 24/12/2013 at Royal Perth Hospital were retrospectively reviewed. Clinical characteristics were determined by reviewing the discharge summary for that admission. Males were further segregated into hs-TnI between 26–33ng/L, and ≥34 ng/L, the internationally proposed male 99^th percentile.

Results

Methods

Modified DSL Gen II, Cobas Elecsys E170 (Roche) and Access 2 (Beckman Coulter) instruments were used to run 39 patient sera in parallel.

Intra-assay imprecision was determined using 20 replicates of patient sera while inter-assay imprecision was determined by running QC material over 20 days.

Results

P-K regression analysis of patient sera (pmol/L) gave the following equations: Access2 = 0.92 DSL Gen II + 1.35

E170 = 0.75 DSL Gen II + 1.29

E170 inter-assay imprecision <2.2%CV across range 6.8–37 pmol/L with intra-assay imprecision <1.1%CV across range 9.8–31.9 pmol/L. Access 2 intra-assay <3.6%CV across range 6.3–98.6 pmol/L with inter-assay imprecision <1.6% for range 8.5–16.0 pmol/L.

Conclusion

Both automated assays were much faster, more precise and less hands-on compared to the ELISA method in use. The slope of 0.75 found in this study for the Roche assay compares well with the slope of 0.81 quoted by Roche. The decision was made to change to the Beckman Coulter Access2 assay to avoid a significant shift in patient results. There remains an urgent need for calibrators traceable to an International Standard for AMH.

CBR. 2015 Aug;36(Suppl):S41.

P64: Development of a Method for the Simultaneous Measurement of Serum Testosterone, 17-Hydroxy Progesterone and Androstenedione by Liquid Chromatograpy – Tandem Mass Spectrometry (LC-MS/MS)

M Henman ^1,^✉, K Waller ¹, J Warner ¹

Abstract

Introduction

Liquid chromatography–tandem mass spectrometry (LC-MS/MS) is ideally suited for accurate, rapid analysis of multiple analytes from a single injection. We developed and validated a sensitive, specific LC-MS/MS method for simultaneous quantitation of testosterone, 17-hydroxyprogesterone (17OHP) and androstenedione. Biochemical assessment of these steroids is important to investigate hyperandrogenism, infertility, polycystic ovarian syndrome, androgen secreting tumours and congenital adrenal hyperplasia. This assay replaced our testosterone immunoassay and by introducing in-house testing for 17OHP and androstenedione, achieved cost-savings for the laboratory.

Methods

After addition of isotope-labeled internal standards, serum samples were deproteinised and extracted with tertiary-methyl-butyl-ether. Supernatants were evaporated and reconstituted. Chromatography was achieved in 15 minutes by RP-C18 column, with a Shimadzu HPLC and ABSciex-QTRAP-5500. The method was validated for linearity, precision, absolute recovery, functional sensitivity, matrix effect and accuracy (against RCPAQAP target and mean values). Patient correlation studies were performed for testosterone (vs Beckman-Access2), 17OHP, and androstenedione (vs reference LCMS/MS).

Results

Testosterone and androstenedione were linear to 125 nmol/L, 17OHP to 375 nmol/L. Within-run precision for all analytes was less than 5% and between-run less than 8.5%. Absolute recoveries were 69–95%. Functional sensitivity was 0.1 nmol/L for testosterone and androstenedione, 0.3 nmol/L for 17OHP. Matrix effects were minimal. RCPA-QAP and patient sample linear-regression comparisons (Passing-Bablok) were y = 1.12x+0.12 and y = 1.30x-0.52 (testosterone), y = 1.06x-0.35 and 1.12x+0.01 (17OHP) and y = 1.05x-0.08 and 1.28x-0.24 (androstenedione). Cost analysis indicated significant per annum savings for the laboratory.

Conclusion

This accurate, rapid, sensitive LCMS/MS assay for simultaneous analysis of testosterone, 17OHP and androstenedione compares well with the RCPA-QAP samples and another LCMS/MS method. Replacing the testosterone immunoassay will mean improved precision and accuracy at low levels especially benefitting female, paediatric and oncology patients. Considerable cost-savings will be achieved by ceasing referrals for 17OHP and androstenedione testing.

CBR. 2015 Aug;36(Suppl):S41–S42.

P65: Review of Sex Hormone Test Requests in a Tertiary Hospital

L Ong ^1,^✉, S Saw ¹, SK Sethi ¹

Abstract

Introduction

Sex hormones testing are commonly requested for assessment of the gonadal-pituitary-hypothalamic axis. However, modern methodologies preclude their use especially in the lower end concentration, hence we reviewed our laboratory’s estradiol and testosterone requests to determine the frequency of such results.

Methods

All estradiol and testosterone requests from April 2014 to April 2015 were reviewed, with the first request considered in patients with repeated requests. Estradiol and total testosterone were run on Beckman Coulter Access. Free testosterone was performed using Diasource radioimmunoassay kit.

Results

There were 6771 estradiol requests for 3072 patients, with median age of 33 and 38 years old for female and male patients respectively, and female:male ratio of 9:1. 178 and 47 patients were below 16 years old or above 65 years old. 289 (93%) male and 1854 (67%) female patients had estradiol levels below the functional sensitivity of 160 pmol/L.

– E2 immunoassays 1^st gen Not suitable for postmenopausal women, males, prepubertal/pubertal children

– Testosterone not for children/women

There were 3513 total testosterone requests for 3038 patients, with median age of 31 and 51 years old for female and male patients respectively, with a female:male ratio of 2:1. 90 (10%) male and 196 (9%) female patients had testosterone levels below the detection limit of 0.35 nmol/L, and 171 patients were below 16 years old. There were 200 free testosterone requests for 171 patients, with median age of 30 and 53 years old for female and male patients respectively, with a female:male ratio of 1:16. 3 (2%) male patients had levels below 0.45 pmol/L.

Conclusion

Despite the Endocrine Society’s recommendation that estradiol immunoassays are not suitable for postmenopausal women, males, prepubertal/pubertal children, a large proportion of our laboratory’s estradiol requests came from these groups. Therefore there’s need to communicate with our clinicians about our assay limitations.

CBR. 2015 Aug;36(Suppl):S42.

P66: Prolactin and Reproductive Hormone Status in Oligomenorrheic and Infertile Females

R Suwal ^1,^✉, AK Nepal ², B Gelal ¹, S Gautam ¹, BKL Das ¹, M Lamsal ¹, S Majhi ¹, N Baral ¹

Abstract

Background

Oligomenorrhea can be a significant problem affecting young women. Oligomenorrhea during the reproductive years may indicate infertility and may cause matrimonial disharmony which is a serious concern in the Asian sub-continent. The present study was designed to assess prolactin, Follicular Stimulating Hormone (FSH) and Luteinizing Hormone (LH) in oligomenorrheic patients in Eastern region of Nepal.

Method

A total of 126 patients from Department of the Obstetrics and Gynecology with complaints of oligomenorrhea, primary or secondary infertility were enrolled in this study. The immunoassay laboratory of the Department of Biochemistry performed measurement of prolactin, LH and FSH by ELISA method (Eliscan, India). Data were analyzed using SPSS ver. 20, p-value <0.05 was considered significant.

Results

The mean age of patients was 24.3±5.9 (range 15–45 years). Most had oligomenorrhea (76.2%) and 23.8% had primary or secondary infertility (pregnancy was excluded). The oligomenorrheic women had median prolactin 698 mIU/mL (interquartile range 500–919), FSH 8.5 mIU/mL (6.2–15) and LH 6.4 mIU/mL (3.6–15). The infertile female group had median prolactin 784 mIU/mL (516–989), FSH 11.0 mIU/mL (8.1–61) and LH 6.3 mIU/mL (3.5–27). Prolactin and LH values were not significantly different between the oligomenorrhea and infertility group (p=0.043).

Conclusion

Our study found no significant differences in median serum prolactin and LH concentrations between oligomenorrheic and infertile women.

CBR. 2015 Aug;36(Suppl):S42.

P67: Development of an Urine Cortisol Assay by Liquid Chromatography-Tandem Mass Spectrometry

R Sahertian ^1,^✉, M Wright ¹, P Stanford ^1,², C White ^1,²

Abstract

Introduction

24hr urinary cortisol is a first line test in the diagnosis of Cushings Syndrome, providing an integrated assessment of cortisol secretion over a 24 hour period. Testing by indirect immunoassay shows poor precision, is vulnerable to interferences and does not assess differences in extraction recovery. Additionally comparisons between immunoassays from different manufacturers demonstrated poor agreement. Our aim was to develop a urine cortisol assay by liquid chromatography/mass spectrometry (LC-MS/MS), suitable to replace the existing immunoassay.

Methods

Initial sample comparisons were performed on Spectra Orion Diagnostica RIA kit, Immulite 2000 and Roche Cobas 8000 immunoassays.

62 urine samples sourced from patient population were analysed by the newly developed LC-MS/MS assay and the existing Immulite 2000 immunoassay. NEQAS Quality Assurance (QA) samples were analysed by LC-MS/MS.

Results

Comparisons between immunoassays demonstrated either good agreement with a large degree of scatter or poor agreement with a smaller degree of scatter. A comparison between the LC-MS/MS method and the Immulite 2000 immunoassay demonstrated imprecision across three pooled human urine samples to be 4.2–5.8% for LC-MS/MS and 13.3–16.8% for Immulite immunoassay. Deming regression analysis, weighted by imprecision, gave “LC-MS/MS” = 1.59(Immulite 2000) −39.5 and a negative bias of 15.8% by Bland Altman analysis. A comparison was performed on samples within the lower end of the calibration curve (28–185nmol/L): The level of agreement on this data set by weighted Deming Regression is “LC-MS/MS” = 1.41(Immulite 2000) −28.4 with a negative bias of 2.6% by Bland Altman analysis. The first distribution (416) of the NEQAS QA scheme (method group n=30) showed a bias between −9.1 and +4.0%.

Conclusion

A rapid LC-MS/MS assay was developed with reduced imprecision compared to the existing Immunoassay, and demonstrated good agreement to the LC-MS/MS group of the UKNEQAS quality assurance scheme.

CBR. 2015 Aug;36(Suppl):S42–S43.

P68: Integrating LC-MS/MS into a Clinical Laboratory: Hands-Free Data Transfer from Sample Handling to Result Entry

R Sahertian ^1,^✉, M Wright ¹, B Patterson ², C Hodgkins ², J Teal ³, C White ^1,⁴

Abstract

Introduction

In a high-throughput liquid chromatography/tandem mass spectrometry (LC-MS/MS) laboratory it is essential to have a means of transferring data from different instruments and the laboratory information system (LIS), ensuring sample identification is maintained throughout and independent of human error. Our aim was to develop an integrated data system from the point of sample preparation, extraction, LC-MS/MS to result entry.

Methods

Barcodes were used to both identify patient samples and 96 well plates, and the order in which they would be processed by automated sample handling technology (TECAN EVO), various LC-MS/MS systems and the LIS. Specialised middleware (XQue) was utilised to ensure hands-free data transfer.

Results

A batch file was created during the sample preparation steps by the sample handler. Data in the batch file included: patient identifier, sample type, lot number/level and position in batch, which was transferred to XQue middleware. 96-well plates containing prepared samples were barcode identified at the point of LC-MS analysis enabling data list upload to the instrument. After batch completion quantitation software processed the data ensuring the following parameters were examined, and flagged: Ion Ratios, Internal Standard variation within defined range, QC values within range, Calibrator accuracy, and sample concentration within calibrator range. All results (including flags) from the batch file were then transferred from LC-MS/MS to XQue, where operators were required to action flagged results prior to transfer to the LIS.

Conclusion

A streamlined, integrated data system was established to facilitate data transfer from the point of sample handling to result entry, ensuring sample identification was maintained throughout the process, and results requiring further examination subjected to scrutiny prior to release.

CBR. 2015 Aug;36(Suppl):S43.

P69: A Pragmatic Approach to Paediatric Reference Intervals for Cerebrospinal Fluid (CSF) Total Protein, Glucose and White Blood Cell Count Using Published Evidence, Clinical Consensus and in-House Verification

CKM Ho ^1,^✉, NWS Tee ¹

Abstract

Introduction

Few hospital laboratories have enough resources to establish their own paediatric reference intervals. Published reference intervals for cerebrospinal fluid (CSF) total protein, glucose and white blood cell count vary widely between studies in the literature and there is no consensus on the best reference intervals to be used in paediatric populations.

Methods

To establish paediatric CSF reference intervals, we employed a three-phase approach. The literature was reviewed and 2.5^th – 97.5^th percentiles were summarized in phase one of the project. During phase two, multiple paediatricians including neurologists, neonatologists and infectious disease specialists were consulted on the most desirable reference intervals to be used based on the literature and their clinical experience. After a consensus on the reference intervals had been achieved, results from at least 20 CSF specimens from each paediatric age group were used to verify the above reference intervals based on both consensus and published evidence. For the purposes of verification, CSF specimens with <200 red blood cells per ul, negative bacterial culture and neither enterovirus nor parechovirus DNA were identified from paediatric patients who also had negative blood and urine bacterial cultures during their hospital admission. Reference intervals were verified based on the methods recommended by the Clinical Laboratory Standards Institute (guideline C28-A3c).

Results

Paediatric CSF reference intervals by consensus were verified as follows.

Total protein (g/L): 0.3–1.2 (<1 month old); 0.2–0.6 (1 to <3 months old); 0.1–0.4 (≥3 months old).
Glucose (mmol/L): 2.0–5.6 (<6 months old); 2.4–4.3 (≥6 months old).
White blood cell count (per ul): 0–20 (<1 month old); 0–10 (1 to <2 months old); 0–5 (≥2 months old).

Conclusion

Using the above approach, we successfully established and implemented paediatric CSF reference intervals that are clinically relevant and locally verified.

CBR. 2015 Aug;36(Suppl):S43.

P71: Thyroid Function Test Reference Intervals During Pregnancy Using the Vitros 5600

E Haworth ^1,^✉, C Chang ², T Yen ³, A Chiriano ¹, S Matthews ¹

Abstract

Introduction

Harmonization of reference intervals (RI) for thyroid function tests (TFT) is restricted by assay variation, but the derivation of ‘method-specific’ TFT RI has been recommended by professional working parties. This study aims to develop evidence-based, Ortho Clinical Diagnostics (OCD) RI for TSH, fT4 and fT3 in pregnancy, since our laboratory is one of few to report TFT results for this method group in the RCPA QAP Endocrine Program.

Objective

Results

Results

There were no false positive or false negative results. All assays apart from phencyclidine marginally over recovered, with recoveries decreasing as the concentration increased. At the cut off concentration, recoveries for barbiturates, oxycodone and propoxyphene were higher than the target concentration by no more than 12%. For phencyclidine, recoveries were 7% lower at the cut off concentration. The coefficient of variation for all drugs over all concentrations was less than 5%. When the manufacturer’s low and high controls were assayed, all drugs apart from propoxyphene under recovered (but by less than 10%). The propoxyphene low control under recovered by 12%. The coefficients of variation for low and high controls of all drugs were less than 7%.

Conclusion

The four immunoassay kits evaluated performed satisfactorily and are suitable as screening tests. Furthermore, these kits have excellent precision around the cut off concentrations recommended by the manufacturer.

PERMALINK

Proceedings of the Australasian Association of Clinical Biochemists’ 53rd Annual Scientific Conference

S1: HBA1C for Diagnosis (Including Pregnancy and in Children)

A McElduff

Abstract

S2: Implementation of HBA1C as a Diagnostic Test in New Zealand

CM Florkowski

Abstract

S3: The Introduction of HBA1C as a Diagnostic Test in Australia

G Jones

Abstract

S5: Time to Retire Microalbumin? - Affirmative

R MacIssac

Abstract

S6 Biochemistry of Phaeochromocytoma

G Eisenhofer

Abstract

S7: Laboratory Testing for Primary Aldosteronism in 2015 – Current Situation and Important New Developments

M Stowasser

Abstract

S8: The Performance/Adequacy of Current Laboratory Assays for GH And IGF1

KKY Ho

Abstract

S9: Getting the Right Answer – The Importance of Traceability

G Jones

Abstract

S10: Review of QAP Activities

K Barancek

Abstract

S12: Diabetic Hyperlipidaemia

D Sullivan

Abstract

S13: Fibrogenesis Biomarkers in Diabetes Complications

SM Twigg

Abstract

S14: Advanced Glycation in Diabetes and Effect of Diet

J Forbes

Abstract

S15: Insulin Assays Fit for Purpose?

P Williams

N Pereria

MS Twigg

T Yen

Abstract

S16: Mechanisms of β-Cell Failure in Type 2 Diabetes

R Laybutt

Abstract

S17: Closing the Loop or Running in Circles? Making Progress

J Gunton

Abstract

Introduction

Conclusion

S18 Assessment of Polycystic Ovary Syndrome

A Joham

Abstract

S21: Steroid Analysis by LC-MS

M Wright

Abstract

S22: Macroprolactin – Current Processes And Recommendations

L Boscato

Abstract

S25: Molecular Diagnostics in Thyroid Cancer

C Woolnough

Abstract

S26: Molecular Testing in Phaeochromocytoma

D Benn

Abstract

Introduction

Methods

Results

Conclusion

S27: Metanephrine Testing - Why, How and When?

G Woollard

M Whiting

Abstract

S28: Quality of HBA1C Measurements in Australia – A Report From the RCPAQAP

P Williams

S Koetsier

V Braniff

G Jones

Proceedings of the Australasian Association of Clinical Biochemists’ 53^rd Annual Scientific Conference