OBJECTIVE: To compare the ELISA (Enzyme Linked Immunosorbent Assay) and the ECLIA (Electro-chemiluminescence Immunoassay) for the estimation of AMH and its correlation with AFC and oocytes retrieved.
DESIGN: Prospective and retrospective observational study.Sample size is 260 in each arm.Duration of the study is from August 2016 to August 2017.
MATERIALS AND METHODS: AMH and FSH estimation is done using ECLIA method by analyzing the serum sample on the Roche-Cobas e411 autoanalyser on a real time basis with a coefficient of variance as 5.5% for level 1 and 5.7% for level 2 for AMH and 2.1% for level 1 and 2.4 % for level 2 for FSH. For the ELISA method, all the data collected retrospectively.
INCLUSION CRITERIA:All patients undergoing IVF treatment . EXCLUSION CRITERIA:All patients with endometriotic/ retention cysts as it may lead to a false impression about the AFC and donor cycles
OUTCOMES MEASURED: Primary: Method of AMH estimation, ECLIA vs ELISA and its better correlation with AFC.Secondary: Method of AMH estimation, ECLIA vs ELISA and its better correlation with number of oocytes retrieved.
STATISTICAL ANALYSIS: Done on MS Excel 2010 and SPSS version 16.Correlation compared with ROC curves.A correlation analysis was performed to determine the degree of agreement between the ELISA and ECLIA kits for AMH estimation using the Kruskall Wallis test.
RESULTS: Post Hoc Tests were carried out for the two groups and shows that when the AFC was compared within the group(AFC- 1-6; 7-19;>/=20)where AMH was measured using ELISA there was no significant difference amongst them showing that AMH was not significantly different between those who had a low AFC and those who had a higher AFC ; result substantiated with Kruskall Wallis test.(p=0.135) However when done with ECLIA - AMH was significantly different , also corroborated by the Kruskall-Wallis test. (p=0.0001)
When group was divided into three sub groups based on the number of oocytes retrieved (1-3;4-14;>/=15)and with AMH being measured using ELISA as a constant the three groups were compared with each other. The AMH again was not significantly different among the three groups and further evaluation by Kruskall- Wallis test also showed no significance.(p=0.885) ; with ELCIA AMH was significantly different amongst them ( P =0.0001)
CONCLUSIONS: ECLIA method of AMH estimation has better correlation with AFC as well as oocytes retrieved than the ELISA method.Cut off value for AMH for poor response in the ECLIA method using the ROC curve analysis is 0.81 with 95% sensitivity and 51% specificity.Cut off value for AMH for hyper response in the ECLIA method using the ROC curve analysis is 6.63 with 63% sensitivity and 95% specificity.ECLIA is an automated method and results can be generated faster with more predictive value.AFC and AMH can be used as a screening test for poor response, but the index IVF cycle remains the best diagnostic tool for poor response.AFC and AMH can be used as a diagnostic test for hyper response.