Wolfler 2005.

Study characteristics
Patient sampling	Primary objective: to evaluate whether endometrial biopsy prior to laparoscopy in symptomatic women to screen for the presence of aromatase by RT‐PCR and IHC combined with select patient characteristics is of value to predict endometriosis Participants: women attending for diagnosis of unexplained infertility, dysmenorrhoea, dyspareunia or chronic pelvic pain scheduled for laparoscopy Selection criteria: exclusion criteria: oestrogen dependent disease other than endometriosis, no endocrine therapy prior to inclusion (GnRH analogues, danazol or oral contraceptives) Study design: observational single‐gate, prospective enrolment and sample collection
Patient characteristics and setting	Clinical presentation: unexplained infertility ‐ 32/48, dysmenorrhoea ‐ 13/48, dyspareunia ‐ 1/48, chronic pelvic pain ‐ 12/48 Age: mean age 32.9 years, range 21‐48 years Number enrolled: 64 women Number available for analysis: 48 women (cycle phase not reported) Setting: tertiary centre ‐ Division of Gynaecological Endocrinology and Reproductive Medicine, Medical University of Vienna Place of study: Vienna, Austria Period of study: not stated Language: English
Index tests	Index test: aromatase mRNA and protein Description of positive case definition by index test as reported: mRNA expression levels by RT‐PCR (Taqman methodology, normalised to internal reference GAPDH gene according to the 2‐ΔΔCt method); protein level ‐ positive staining on IHC (detection by ChemMate Antibody Detection Kit (Dako); microscopical examination of 6 section per slide); laboratory techniques described; threshold pre‐defined for IHC Examiners: IHC ‐ 3 independent trained physicians who were unaware of the patients' history Interobserver variability: not stated
Target condition and reference standard(s)	Target condition: endometriosis Prevalence of reference condition in the sample: n/N = 25/48 (52%): stages I‐IV, numbers not specified; controls 23 Reference standard: laparoscopy + histology Description of positive cased definition by index test as reported: laparoscopic visualisation followed by histopathological assessment; rAFS classification; visual diagnosis that could not be confirmed by histopathology was considered as negative Examiners: not stated
Flow and timing	TIme interval between reference standard and index test: tissue samples were collected prior to laparoscopy Withdrawals: 16 patients were excluded from the analysis: 7 patients ‐ poor quality samples, 2 patients ‐ insufficient RNA extraction, 2 patients ‐ insufficient sample for duplicates, 5 patients ‐ were using COCs
Comparative
Notes	Conclusion: Screening for eutopic endometrial aromatase in combination with clinical data could be of discriminative value in the prediction of disease. Comments: The reported diagnostic estimates of predictive model based on endometrial sample and clinical data are beyond the scope of this review. For aromatase mRNA there was statistically significant difference between the groups, but no data available to construct 2 × 2 tables; 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?	Unclear
Did the study avoid inappropriate exclusions?	Yes
Was a 'two‐gate' design avoided?	Yes
		Unclear	Low
DOMAIN 2: Index Test All tests
Were the index test results interpreted without knowledge of the results of the reference standard?	Yes
If a threshold was used, was it pre‐specified?	Yes
Was a menstrual cycle phase considered in interpreting the index test	No
		High	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?	No
		High