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Objective A novel corona virus(SARS-CoV-2) nucleic acid detection kit was developed and performance had been optimized for large-scale population screening in China. The effect of mixed detection of multiple samples was analyzed aiming at providing theoretical guidance for clinical use. Methods The primers and probe sequences for ORF1ab and N announced by China Center for Disease Control and prevention and the primers and probe sequences for RNase P as control gene were synthesized. Single variable optimization and orthogonal experiment were used to get the best concentration ratio of primers and probes for ORF1ab, N and RNase P in fluorescent RT-qPCR detection system. The accuracy, specificity and sensitivity of the kit were investigated. The positive samples and negative samples were collected and mixed according to different proportions, and the SARS-CoV-2 was detected by this kit and the commercial reference kit respectively. The detection concentration for different targets and the amplification Ct value were compared. Results The positive coincidence rate, negative coincidence rate and specificity of the kit were 100%, respectively。And the sensitivity was 1×103 copies/mL. The detection sensitivity of this kit for ORF1ab target gene was 1/20, higher than that of reference kit (1/10), and the amplification Ct value with same template concentration was lower than that of reference kit (P＜0.05). Conclusions This kit has great advantages in screening multi samples detection of new corona virus nucleic acids because of high sensitivity to ORF1ab target gene, and suitable for in large-scale population.