Aims: To diagnose male infertility, researchers examine sperm deoxyribonucleic acid (DNA) damage. The present study aims comparing different DNA damage methods like Free DNA estimation, Comet assay and evaluation of γH2AX levels to rapidly detect infertility among sperm samples. 

Materials and methods: A total of 151married volunteers of 35 o 45 age were selected and semen samples were collected every year for 5 years. Persons were classified into two groups based on confirmed fertility and reduced fertility. The samples were processed using γH2AX quantization by immune blotting, an alkaline comet assay and free DNA estimate using UV 260nm measurement and diphenylamine (DPA) method.

Results: The findings of the γH2AX quantization by immune blotting, DNA damage by comet assay and the estimate of free DNA differed between the two groups. All three methods responded similarly in two tested groups. Compared to the fertility groups, the unproven fertile groups had higher γH2AX levels, length of comet tails and free DNA content. γH2AX was early detected before free DNA and comet assay but its presence also noted in radiation exposed and hypoxic patients . Free DNA was found in aged and infertile persons. Hence γH2AX assay and DNA damage coupled can be used for infertility test.

Conclusion: DNA damage might cause male infertility. This DNA damage can be assessed by γH2AX quantization by immune blotting, DNA damage by comet assay and the estimate of free DNA

Clinical significance: γH2AX quantization by immune blotting along with DNA damage by comet assay or the estimation of free DNA might be used to profile male infertility. 

Chand Pasha is an Assistant Professor and Head of the Department of Microbiology at Nizam College, Osmania University, Hyderabad, India. He earned his M.Sc. in Microbiology and Ph.D. in Molecular Microbiology from Osmania University and also holds postgraduate diplomas in Patent Laws from NALSAR University and the World Intellectual Property Organization (WIPO). With extensive experience in microbiology research and higher education, he has been actively involved in teaching, research, and academic administration. His research interests include molecular microbiology, biotechnology, and intellectual property in life sciences. He has received several recognitions, including Junior and Senior Research Fellowships from the Department of Biotechnology, Government of India, and Best Poster Awards at national scientific conferences.