Infertility is a serious reproductive health problem affecting family well-being and harmonious development in our country; globally, around 15% of heterosexual couples are unable to conceive their children without assistance. Assisted reproductive technology (ART) and precision medicine are key to resolve this problem, in which evaluation of sperm quality is necessary and important. However, the world health organization (WHO) laboratory manual for the examination and processing of human semen (fifth edition, 5th) just provides a rough analysis to evaluate human sperm, which brings inconsistency in clinic. How to precisely evaluate human sperm is an imminent and very challenging problem. In this study, a method in terms of protamine (PRM) pattern and value to evaluate human sperm was established. The primary result obtained by using of this new method is consistent with clinical outcome and better than that obtained by using WHO standard manual (5th). Thus, this study is valuable to ART and precision medicine, which can improve ART success rate and safety in clinic.