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Quadruplex Real-Time qPCR Assay for the Simultaneous Assessment of Total Human DNA, Human Male DNA, DNA Degradation and the Presence of PCR Inhibitors in Forensic Samples: A Diagnostic Tool for STR Typing

NCJ Number
Forensic Science International: Genetics Volume: 2 Issue: 2 Dated: March 2008 Pages: 108-125
Date Published
March 2008
18 pages
This paper reports on the developmental validation of a quadruplex real-time qPCR assay (analysis) for the concurrent assessment of the total human DNA, human male DNA, DNA degradation and the presence of PCR inhibitors in forensic samples.

The ability to accurately determine the quantity of DNA present in a forensic sample and simultaneously determine the most appropriate typing system for each sample may result in substantial increase in laboratory efficiency and a more prudent use of limited samples. The nuTH01-nuSRY-nuCSF-IPC quadruplex qPCR assay has been shown to provide suitable quantifications for both total human and total human male DNA while providing qualitative information about the level of degradation and the presence of PCR inhibitors. The results from the nuTH01-nuSRY-nuSRY-nuCSF-IPC quadruplex qPCR assay can also be used to confirm the results of short tandem repeat (STR) typing, which may present unnecessary reanalysis of degraded or inhibited samples that have already yielded the maximum amount of information available. The increasing number of DNA typing system and decreasing amount of DNA per evidence item submitted to forensic laboratories has driven the development of several real-time quantitative PCR (qPCR) analyses capable of simultaneously estimating total human DNA and male DNA or of estimating total human DNA and extent of DNA degradation. This paper describes the quadruplex real-time qPCR assay developed to simultaneously assess total human DNA, human male DNA, DNA degradation, and PCR inhibitors in forensic samples. Tables, figures, references

Date Published: March 1, 2008