The selection of Y chromosome microdeletion detection methods based on seminal analysis results: a comparison of high-throughput sequencing and fluorescence quantitative polymerase chain reaction (qPCR) applications

Abstract

Background: Infertility significantly impacts numerous couples worldwide, and male infertility is a common contributing factor. Y chromosome microdeletions are potential genetic causes of male infertility. However, due to the lack of comparative studies based on semen analysis results, we have difficulty selecting an appropriate method for detecting Y chromosome microdeletions. This study aims to compare the application of high-throughput sequencing and fluorescence quantitative polymerase chain reaction (qPCR) in different types of infertility patients.

Methods: This study used high-throughput sequencing [next-generation sequencing (NGS)] and fluorescence qPCR methods to detect Y chromosome microdeletions in two groups: one with azoospermia and another with oligoasthenoteratozoospermia (OAT), characterized by reduced sperm count and motility.

Results: The results showed that NGS identified cases of Klinefelter syndrome (congenital bilateral absence of the vas deferens) that were not detected by qPCR in the azoospermia group. In the OAT group, high-throughput sequencing found a b2/b3 deletion of 1.80 Mb, while qPCR did not detect it. Conversely, qPCR identified an AZFd deletion in the OAT group, missed by high-throughput sequencing due to inadequate target region coverage.

Conclusions: These research findings are significant for guiding personalized treatment of male infertility patients and provide valuable references for further exploration of the association between Y chromosome microdeletions and male infertility.

Keywords: Y chromosome microdeletion; fluorescence quantitative polymerase chain reaction (fluorescence qPCR); high-throughput sequencing; infertility; seminal analysis.

Figure 1

High-throughput abnormality analysis results. (A) In the azoospermia group, Y chromosome microdeletion analysis through high-throughput sequencing showed normal results, with no overall abnormalities detected, indicating Klinefelter syndrome patients. (B) In the oligoasthenoteratozoospermia group, case 1 exhibited a deletion in the AZFc region with a length of 3.50 Mb. (C) In the oligoasthenoteratozoospermia group, case 2 exhibited a deletion in the b2/b3 region with a length of 1.80 Mb.