四引物扩增受阻突变体系分析少精、无精症患者白细胞MLH3基因C2531T突变

Detection the MLH3 gene C2531T mutation in oligospermia and azoospermia by 4P-ARMS-PCR

目的分析错配修复蛋白MLH3基因C2531T突变与少精、无精症的关系。方法以MLH3C2531T为例建立四引物扩增受阻突变体系(4P-ARMS-PCR)方法;分别用4P-ARMS-PCR和PCR-RFLP检测81例少精、无精症患者和80例正常对照者MLH3C2531T;随机抽取20%标本测序进行验证。结果MLH3C2531T的CT+TT基因型在患者中有明显的流行,且MLH3C2531T的T等位基因与疾病存在相关性(P<0.01);PCR-RFLP检测结果与4P-ARMS-PCR检测结果完全一致,测序结果相同。结论4P-ARMS-PCR可快速、简便、准确检测基因单核苷酸突变(SNP);错配修复蛋白MLH3C2531T多态性与少精、无精症存在相关性。

Objective Analysing the potential relationship of mismatch repair protein MLH3 gene C2531T mutation with oligospermia and azoospermia cases. Methods Using MLH3 C2531T as an example to establish a method of tetra-primer amplification refractory mutation system-PCR （4P-ARMS-PCR） ;We chose 81 cases of oligospermia, azoospermia patients and 80 normal controls, detecting MLH3 C2531T both using 4P-ARMS-PCR and PCR-RFLP; randomly to select 20% samples to sequence, Results The results suggested higher prevalence of CT ＋ TT genotypes in patients as compared to the controls and association of T allele with disease（ P 〈 0. 01 ） ; PCR-RFLP results, 4P-ARMS-PCR results and sequence results are fully consistent. Conclusion 4P-ARMS-PCR can be a fast, easy-operating and accurate technology for detection of gene mutation of single nueleotide polymorphism （SNP） ; MLH3 C2531T polymorphism is related to oligospermia and azoospermia.