Design of a SNaPshot assay for simple and cost-effective detection of six variant related to recurrent pregnancy loss

Backgrounds: Applying karyotyping as conventional approach is still mainly applicable inlaboratory but a considerable proportion of RPL (recurrent pregnancy loss) is related to singlegenealteration such as mutation, insertion and etc. Our aim in this study was to design anddevelopment a genetically panel using SNaPshot method besides karyotyping.Materials and Methods: Six specific primers were designed to amplify target region involvedin pathogenesis of RPL through multiplex PCR method. Specific single base extension (SBE)designed in order to mini-sequencing six mutation site by the SNaPshot simultaneously. TheSBE products were electrophoresed in an ABI ۳۱۳۰xl Genetic TM Analyzer) using POP-۴polymer and ۳۵ cm capillary arrays. The GenMapper v۳.۲ software was used to analyze theresulting electropherograms.Results: Our pilot study six mutation sites in ۱۶ women with RPL were previously genotyped byreal-time PCR, were used to test the accuracy and reproducibility of multiplex SNaPshot assays.The results were compared with the previously analyzed types.Conclusion: SNaPshot technique is specific, accurate and inexpensive approach to customizedgenetically panel and monitoring of frequent pathogenic mutation in RPL.

کلیدواژه ها:

Recurrent pregnancy loss ، Multiplex ، SNaPshot ، Mini-sequencing ، Genetically panel