A Preclinical and Clinical Trial Evaluation for an in vitro Diagnostic Kit of Endometriosis (I)

Endometriosis occurs when shed endometrium consisting of endometrial stroma and epithelial cells form the female reproductive tract grows at extrauterine sites. It is a benign but aggressive disease which could be classified into four stages: minimal, mild, moderate, and severe. Endometriosis is affecting 2-10% of woman of reproductive age worldwide and considered to be a major reason for infertility. It is suggested that early detection of endometriosis and treat with GnRH agonist to reduce the level of estrogen could increase pregnancy rate for minimal-mild patients. Unfortunately, the gold standard for the confirmation of endometriosis remains a surgical procedure, laparoscopic examination, since non-invasive approach such as ultrasound, MRI and blood tests have not been proved reliable. The presenting invention relates a non-invasive diagnosis method of endometriosis by detecting a new biochemical marker in serum or peritoneal fluid. Alpha-1 antitrypsin (A1AT) was selected to be endometriosis biomarker as found increased in serum of endometriosis women by proteomic analysis. Immuno-dot blotting was performed thereafter to measure A1AT expression level in relation to endometriosis. The data shows that women with endometriosis have significantly higher A1AT expression than control group. Moreover, A1AT shows cumulative expression in the progression of endometriosis. Importantly, a decrease of the concentration of A1AT after being treating with GnRH agonist is observed. It suggests that A1AT has a great potential for endometriosis diagnosis and prognosis. It is known that some clinicians are adopting CA125 as a help of diagnosing endometriosis. It should be note that the increase of CA125 usually related to several physiological and pathologic conditions. Moreover, CA125 shows almost no increase in the early stage of endometriosis. In general, the specificity and sensitivity of CA125 for diagnosing endometriosis is relative low, only 92% and 15% respectively. The practical use of CA125 as a marker therefore, is limited. As a comparison, the presenting biomarker, A1AT, is characterized and examined. Over 200 women serum sample were tested which revealed that the A1AT have high specificity and sensitivity (over 93% and 90% respectively).It should be noted that the invention team have also developed two specific monoclonal antibodies for A1AT with high competitiveness than other commercially available antibody products. The homemade A1AT antibodies, 2A7 and 2C8 can detect serum A1AT with 1:100,000 dilution and the first generation of serum diagnostic kit for endometriosis therefore is under development.In this study, the invention team in collaborate with the Research Center for Clinical Tral in Taipei Medical University Hospital would like t’o collect 881 cases for a ramdomized, blind clinical trial to evaluate the current developed “New” A1AT in vitro serum diagnostic kit for screening high risk patients with endometriosis. By comparing the CA125 index and an A1AT test for other disease (Simens) on the specificity and sensitivity of the diagnostic test for endometriosis, the accuracy and potential of the“New” A1AT in vitro serum diagnostic kit for endometriosis can be learned and evaluated.