Agic 2008.

Study characteristics
Patient sampling	Primary objective: to investigate the combination of CCR1 mRNA, MCP‐1, and CA‐125 protein measurements in peripheral blood as a diagnostic test for endometriosis and to study the possible use of these markers in the peripheral blood of patients with adenomyosis Participants: patients who underwent laparoscopy for various indications Selection criteria: Inclusion criteria: no endocrine therapy for at least 3 months; exclusion criteria: suspected or ascertained diagnosis of malignancy, pregnancy, menopausal age or refusal to participate in the study Study design: cross‐sectional, two‐gate design, prospective collection of samples
Patient characteristics and setting	Clinical presentation: endometriosis group ‐ dysmenorrhoea, dyspareunia, chronic pelvic pain and infertility; 12 women with known history of endometriosis; controls ‐ undergoing surgery for subserosal leiomyomata or tubal ligation Age: reproductive age Number of participants enrolled: 151 women (11 women with adenomyosis were enrolled and analysed separately ‐ not considered in this review) Number of participants available for analysis: 151 women (all in follicular cycle phase) Setting: Department of O&G, University of Schleswig‐Holstein Place of study: Luebeck, Germany Period of study: not stated Language: English
Index tests	Index test: CA‐125, CCR1, MCP‐1 Details of the index test procedure as stated: CCR1 expression detected by RT‐PCR (SuperScriptTM II RT, SYBR Green MM, normalised to HPRT housekeeping gene); MCP‐1 levels detected by using a commercially available ELISA kit (R&D Systems, GmbH, Germany) with assay sensitivity of 5 pg/ml; CA‐125 level detected by using a commercially available electro‐chemiluminescent immunometric assay (ECLIA, Roche Diagnostics GmbH, Germany) with assay sensitivity of 0.6 IU/ml; all the experiments were repeated x 3 times; the test was considered positive for endometriosis if at least one of the markers was above the threshold; sample processing and experiments described Threshold for positive result: CCR1/HPRT > 1.16, MCP‐1 > 140 pg/ml, CA‐125 > 50 IU/ml‐ all pre‐specified Examiners: no information provided; unclear if blinded to the result of reference standard Interobserver variability: for MCP‐1 the intra‐ and interassay CV was 2.5% and 4.5%
Target condition and reference standard(s)	Target condition: endometriosis Prevalence of target condition in the sample: n = 102/151 (68%): stage I‐II 37, stage III‐IV 65; controls n = 49 Reference standard: laparoscopy N = 151 (100%) + histology Description of positive case definition by reference standard as reported: visual inspection, histological diagnosis; staging according to the rAFS classification Examiners: no information provided
Flow and timing	Time interval between index test and reference standard: blood samples were obtained 24 hours prior to anaesthesia and laparoscopy Withdrawals: none
Comparative
Key conclusions by the authors	The results imply the potential use of CCR1 mRNA, MCP‐1, and CA‐125 protein measurements for the diagnosis or exclusion of endometriosis
Conflict of interest	Not reported
Notes	The reported estimates for diagnosis of adenomyosis are not presented in this review The reported diagnostic estimated per severity of endometriosis are not presented in this review
Methodological quality
Item	Authors' judgement	Risk of bias	Applicability concerns
DOMAIN 1: Patient Selection
Was a consecutive or random sample of patients enrolled?	No
Did the study avoid inappropriate exclusions?	Yes
Was a 'two‐gate' design avoided?	No
		High	High
DOMAIN 2: Index Test All tests
Were the index test results interpreted without knowledge of the results of the reference standard?	Unclear
If a threshold was used, was it pre‐specified?	Yes
Was a cycle phase considered in interpretation of the result of index test?	Yes
		Unclear	Low
DOMAIN 3: Reference Standard
Is the reference standards likely to correctly classify the target condition?	Yes
Were the reference standard results interpreted without knowledge of the results of the index tests?	Yes
		Low	Low
DOMAIN 4: Flow and Timing
Was there an appropriate interval between index test and reference standard?	Yes
Did all patients receive the same reference standard?	Yes
Were all patients included in the analysis?	Yes
		Low