Efficient evaluation of the primordial follicle pool (PFP) of mammalian models is an essential subject in biomedical research relating to ovarian physiology and pathogenesis. Our recent study has identified a gene signature includingSohlh1,Nobox,Lhx8,Tbpl2,Stk31,Padi6, andVrtnstrongly correlated with ovarian reserve by using bioinformatics analysis. Aimed to investigate the validity of these candidate biomarkers for evaluating the PFP, we utilized an OR comparison model to decode the relationship between the numbers of PFP and candidate biomarkers in the present study. Our results suggest that these biomarkersSohlh1, Nobox, Lhx8, Tbpl2, Stk31, Padi6, andVrtnpossess independent potential to evaluate the number of the PFP. And the combination ofSohlh1andLhx8can be used as the optimal biomarkers for rapid assessment of the PFP in the murine ovary. Our findings provide a new perspective for evaluating the PFP of the ovary in animal studies and the clinic.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12958-023-01097-3.