Development Of Multiplex Pcr To Assess Male Infertility

Abstract

The second most populous country on earth, India accounts for more than 17 per cent of world population. Rather population growth is a major concern, yet infertility is a devastating health problem. According to WHO, infertility is the inability of the female partner to attain pregnancy after one year or more of the unprotected sex. Infertility affects about 15% of the couple of which half of the reproductive problems are associated with the male counterpart (Agarwal et al, 2015). Causes of infertility in men include a variety of things but can be grouped into a number of genetic and non genetic categories. Non-genetic conditions include hypogonadotrophic hypogonadism, testicular maldescence, structural abnormalities of the male genital tract, genital infections, previous scrotal or inguinal surgery, varicoceles, chronic illness and medication (Crosignani et al., 1993). Genetic causes include chromosomal abnormalities [Klinefelter syndrome (XXY) and specific translocations, point mutations in the androgen receptor and the cystic fibrosis transmembrane conductance regulator (CFTR) gene], Y chromosome microdeletions, mitochondrial DNA (mtDNA) mutations, monogenic disorders, multifactorial disorders and endocrine disorders of genetic origin. Genetic causes of infertility are an important etiological factor leading to irreversible partial or complete spermatogenetic arrest among which Y chromosome microdeletions receives prime focus. The microdeletions that occur in the AZF [AZoospermic Factor] region of the long arm of Y chromosome affect genes that are involved in spermatogenesis. There are three recurrently deleted nonoverlapping subregions in the proximal, middle and distal Yq11 regions in the deleted interval designated as AZFa, AZFb and AZFc (Kent First et al., 1999).  It has been identified in 12.9% of south Indian infertile men (Poongothai et al., 2008). Still now no compact method is available for general public for Y chromosome microdeletion analysis (YCM) in India. European Academy of Andrology (EAA) recommends that 90 % of YCM can be detected by the use of 2 STS markers for each AZF loci. In the present study, method to assess male infertility in Indian population using multiplex PCR employing 3 STS markers for each AZF loci was devised.