目的 通过对不同受孕能力精子外显子的分析,寻找并验证特异性外显子作为精子受孕能力的生物标记物。 方法 基于二代测序数据进行生物信息学分析,寻找特异性外显子并设计引物。各取8份高、低受孕能力精液标本,提取精子RNA反转录后进行real time q-PCR验证外显子的表达效率,筛选表达差异恒定的精子外显子作为生物标记物。各取10份高、低受孕能力精子标本,用筛选后的外显子引物进行real time q-PCR验证。 结果 生物信息学分析得到31个候选精子外显子,从31个候选外显子中筛选出9个表达差异恒定的精子外显子GAPDHS、HSF2BP、HSPA1L、ADAM21、SPEM1、WBP2NL、DDX20、TSGA10、PGK2;real time q-PCR验证结果显示,在高、低受孕能力精液标本中这9种精子外显子表达差异明显。 结论 初步确定,差异表达恒定的九种外显子可作为评估精子质量的生物标记物。

Objective To find and verify specific exons as biomarkers of sperm fertility by analyzing sperm exons with different fertility ability.Methods Based on the second generation sequencing data, bioinformatics analysis was conducted to find specific exons and design primers. We obtained 16 semen samples, 8 of high and the other 8 of low fertilizing ability, after the sperm RNAs were extracted and reverse-transcribed, real time q-PCR was performed to verify the expression efficiency of exons, and the sperm exons with constant expression difference were selected as biomarkers. 10 high and 10 lowfertility ability sperm samples were taken for real time q-PCR verification with screened exon primers. Results Thirty-one sperm exons were obtained by bioinformatics analysis, and 9 sperm exons with constant expression differences were selected from the 31 candidate exons, including GAPDHS, HSF2BP, HSPA1L, ADAM21, SPEM1, WBP2NL, DDX20, TSGA10 and PGK2. The results of real time q-PCR verification showed that the exons of these 9 sperm were significantly different in the semen samples with high and low fertility ability. Conclusion Nine exons with constant differential expression can be used as biomarkers to evaluate sperm quality.