Methods and test kits for determining male fertility status

A method comprising:

• exposing, in vitro, a sperm sample from a male to capacitating conditions;
• fixing the in vitro capacitated sperm sample with a fixative for at least 0.5 hour;
• treating the fixed in vitro capacitated sperm sample with a labeling molecule for GM1 localization patterns, wherein the labeling molecule has a detectable label;
• identifying more than one GM1 labeled localization patterns in the labeled fixed in vitro capacitated sperm sample, said GM1 labeled localization patterns being selected from the group consisting of an apical acrosome (AA) GM1 localization pattern, an acrosomal plasma membrane (APM) GM1 localization pattern, a Lined-Cell GM1 localization pattern, a post acrosomal plasma membrane (PAPM) GM1 localization pattern, an apical acrosome/post acrosome (AA/PA) GM1 localization pattern, an intermediate (INTER) GM1 localization pattern, an equatorial segment (ES) GM1 localization pattern, and a diffuse (DIFF) GM1 localization pattern;
• assigning the apical acrosome (AA) GM1 localization pattern and the acrosomal plasma membrane (APM) GM1 localization pattern to a capacitated state;
• determining a fertility threshold corresponding to a ratio of (i) the number of GM1 labeled localization patterns assigned to the capacitated state and (ii) a total number of GM1 labeled localization patterns;
• comparing the fertility threshold to a reference value;
• and
• determining a fertility status of the male based on said comparison identifying the male subject for assisting reproduction treatment.