Objective: To evaluate the role of auto-antibodies in Chinese patients with primary and secondary infertility in routine clinical practice. Methods: We reviewed the medical records of infertile Chinese patients of either gender (>= 18 year old) who were failed to conceive after 12 months of regular sexual intercourse without the use of contraception, and visited at department of Gynaecology and Obstetrics, Renmin Hospital of Wuhan University, China between January 2004 to December 2014, and undergone auto-antibody estimation along with other routine antenatal screening tests. Immunological assay was performed using dot immuno-gold filtration assay. Positive case was compared among patients with primary and secondary infertility. Results: Our chart review study showed that the positive case of Anti-Sperm Antibody (ASAb) was significantly higher in female with primary than male (p<0.05). In female patients with primary infertility, all other auto-antibodies (anti-Endometrium Antibody (EMAb), Anti-Ovary Antibody (AOAb), Anti-Zona Pellucida Antibody (AZP)) and Anti-Cardiolipin Antibody (ACA) were significantly higher compared to patients with secondary infertility (p<0.05). Moreover, positive case of all auto-antibodies was significantly higher among female with primary fertility than secondary fertility and control groups. Conclusion: Our study results suggested the involvement of auto-antibodies related to reproduction system in development of primary infertility. Auto-antibodies related to reproduction were significantly higher in patients with primary infertility, this implicated involvement of auto-immunological factors in development of infertility.