Abstract: Objective Analysing the association of mismatch repair protein MLH3 gene C2531T mutation in 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 date 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 rapid, simple and accurate detection of gene mutation of single nucleotide polymorphism (SNP); MLH3 C2531T polymorphism is associated with oligospermia and azoospermia.

Key words: 4P-ARMS-PCR, oligospermia, azoospermia, MLH3