Application of Real-time PCR technique to detect AZF microdeletions in infertile men without sperm

Abstract

Application of Real-time PCR technique to detect AZF microdeletions in 30 infertile men without sperm. Method: 30 peripheral blood samples were extracted with DNA, Real-time PCR technique using 21 target sequences to detect microdeletions of AZFabcd, including 6 primer pairs of AZFa (sY84, sY86, sY82, sY1064, sY1065, sY88), 6 primer pairs of AZFb/d (sY127, sY134, sY105, sY121, sY143, sY153), 7 primer pairs of AZFc (sY254, sY255, sY1191, sY1192, sY1196, sY1291, sY160), and two internal STSs ZFY/X, SRY. The results of the control analysis by the multiplex PCR technique revealed 8 basic STSs (sY84, sY86, sY127, sY134, sY254, sY255, sY153, PB2), and the capillary electrophoresis with AZF expansion (10 extended STSs) exhibited 4 primer pairs of AZFa (sY82, sY83, sY88, sY1065), 3 primer pairs of AZFb (sY105, sY121, sY1192), 3 primer pairs of AZFc (sY1191, sY1291, sY160), and two internal STSs ZFY/X and SRY. These techniques were used for diagnosis. Results and conclusions: there were 40% (12/30) of patients with microdeletions in the AZF, basic microdeletions and extention microdeletions were 16.7% and 83.3%, respectively. In extention locations, the major microdeletions were sY1291, sY1191 (AZFc) and sY1192 (AZ Fb).