(SNPs) and infertility in Chinese males using multi-analyte suspension array (MASA). A total of 196 male patients with azoospermia or severe oligospermia (sperm density <5x106/ml, non-obstructed) who had a normal karyotype and no azoospermia factor microdeletions were recruited, along

The aim of the present study was to investigate the association between two single nucleotide polymorphisms (SNPs) and infertility in Chinese males using multi-analyte suspension array (MASA). A total of 196 male patients with azoospermia or severe oligospermia (sperm density <5x106/ml, non-obstructed) who had a normal karyotype and no azoospermia factor microdeletions were recruited, along with 40 healthy, fertile males as controls. Two SNPs of the deleted in azoospermia-like (DAZL) gene, SNP260 and SNP386, were genotyped by allele‐specific primer extension (ASPE) combined with MASA technology. The SNP260A>G and SNP386A>G mutations were found in the males with infertility. The SNP260, but not the SNP386, mutation was detectable in the control group. The mutation rates in the controls and patients were 2.5 and 3.06% for SNP260, and 0 and 2.04% for SNP386, respectively. A χ2 analysis did not identify any significant differences in the frequency of either mutation between the fertile and infertile males. In conclusion, the combination of ASPE and MASA methods for SNP genotyping was high‐throughput, accurate and cost‐efficient. The method was applied to detect SNP polymorphisms in the DAZL gene; and neither the A260G nor the A386G polymorphism of DAZL appeared to be involved in male infertility in the Chinese population. Introduction Male infertility is considered to be associated with azoospermia factor (AZF), one of several genes located in the Yq11 chromosomal region. The most common mutations identified in AZF are microdeletions in AZFc (60%) (1,2). The members of the deleted in azoospermia (DAZ) gene family are the primary regulators of proliferation in early germ cells and are important candidate genes for male infertility in the AZFc region (3). There are three genes in the DAZ gene family: DAZ, DAZL and BOULE. The DAZ-like (DAZL) gene is located in the 3p24 chromosomal region and is an autosomal homolog of DAZ; 83% of the cDNA coding regions in DAZ and DAZL are similar (4-6). A previous study suggested that DAZL is important in the sperm production process (7). However, there are conflicting studies regarding whether DAZL mutations impact male infertility. Following completion of the human genome project, single nucleotide polymorphisms (SNPs) have been intensively investigated. As genetic markers, SNPs have been associated with pharmacogenomics and disease susceptibility. Two studies (8,9) have described A>G transitions at the SNP260 and SNP386 nucleotide positions of DAZL, and SNP386 was found to be associated with susceptibility to spermatogenic failure in the Taiwanese population. However, studies in other countries, including Italy, India and Japan, have indicated that there is not an association between the two SNPs and spermatogenic impairment (10-13). Thus, the role of the SNP260A>G and SNP386A>G transitions in male infertility is controversial (12-16). To the best of our knowledge, there have been no studies regarding DAZL SNPs in the Chinese population.Therefore, in the present study, the distribution of the DAZL A260G and A386G SNPs in Chinese males was investigated. The methods for SNP genotyping are advancing. The available technology includes: Restriction fragment length polymorphism (RFLP), Taqman, high performance liquid chromatography and single-strand conformation polymorphism analysis (17). The most common methods used currently in China to detect SNPs are RFLP and DNA sequencing; however, these techniques are limited in their applications due to the high costs in time and resources. In the present study, a high-throughput, low-cost and low‐consumption method that combined allele‐specific primer extension (ASPE) and multi-analyte suspension array (MASA) Analysis of DAZL SNP260 and SNP386 in infertile Chinese males using multi-analyte suspension array YIJIAN ZHU, MINGFU MA, LING WAN, DANYAN ZHANG, LETIAN ZHAO, LI WEI and LIANBING LI Key Laboratory of Birth Defects and Reproductive Health of The National Health and Family Planning Commission (Chongqing Population and Family Planning Science and Technology Research Institute), Chongqing 400020, P.R. China Received September 24, 2013; Accepted February 11, 2014 DOI: 10.3892/mmr.2014.2634 Correspondence to: Dr Lianbing Li, Key Laboratory of Birth Defects and Reproductive Health of The National Health and Family Planning Commission (Chongqing Population and Family Planning Science and Technology Research Institute), No. 18 Honghuang Road, Jiangbei, Chongqing 400020, P.R. China E-mail: [email protected] Abbreviations: MASA, multi-analyte suspension array; SNP, single nucleotide polymorphism; ASPE, allele-specific primer extension; DAZL, deleted in azoospermia-like