[Validation and Forensic Application of a Domestic Human DNA Quantitative Detection Kit].

Abstract

Objectives: To verify the efficacy of a domestic human DNA quantification kit based on real-time fluorescence quantitative PCR in detecting the total human DNA concentration, male DNA concentration in mixed male/female DNA samples, the degree of DNA degradation and inhibitor tolerance.

Methods: Samples with different concentrations, different male/female ratios, different concentrations of inhibitors, and different degradation degrees were tested using the domestic human DNA quantification kit based on real-time fluorescence quantitative PCR. This kit was compared with a similar product on the market and was applied to the detection of DNA from real cases.

Results: This human DNA quantification kit can effectively detect human DNA as low as 0.001 65 ng/μL, and 6.25 pg/μL of male DNA in mixed samples with a male-to-female ratio of 1∶15 000. Even when the sample contains as high as 400 ng/μL of humic acid or 1 000 μmol/L of hemin alone, the DNA concentration can still be accurately detected. The degradation index can effectively characterize the degradation degree of the sample. This kit has been successfully applied in forensic practice.

Conclusions: This human DNA quantification kit is accurate and reliable in detection. It can accurately reflect the degradation of DNA and inhibitor tolerance. It has good performance in quantitative accuracy, determination of the male/female ratio in mixed samples, and inhibitor tolerance. It has application potential in forensic case examination.