title: An examination of the utility of a nuclear DNA/mitochondrial DNA duplex qPCR assay to assess surface decontamination of hair.
abstract: The goal of this study was to compare two commonly used methods for the surface decontamination of human hair shafts, and to evaluate the use of a duplex real-time qPCR assay to assess decontamination effectiveness for the purpose of mitochondrial DNA typing. Hair shafts of known mitochondrial DNA haplotype were coated with undiluted saliva, semen or blood, each of known mitochondrial haplotype distinct from the test hair. Surface decontamination was conducted by enzymatic treatment with Terg-a-zyme™ and by chemical treatment with dilutions of sodium hypochlorite (NaClO, bleach). Following DNA extraction, a duplex (nuclear and mitochondrial DNA) real-time qPCR assay was used to quantify mitochondrial DNA and to test for surface contamination by quantifying the exogenous nuclear DNA not removed from the hair shaft. The NaClO treatment was found to be more effective for removing surface contamination than the Terg-a-zyme™ treatment, and it was procedurally simpler to implement, resulting in a significant savings of sample processing time. Exposure to 3% NaClO for up to two minutes had no detrimental effect on quantity or typing of the mitochondrial DNA belonging to the hair. In addition, we demonstrated that the duplex real-time PCR assay is a convenient early-warning diagnostic method for the detection of the presence of external DNA contamination, providing an assessment of the purity of the sample prior to embarking on further analysis by more laborious mitochondrial DNA typing methods.
authors.affiliation: Jan Bashinski DNA Laboratory, Bureau of Forensic Services, Department of Justice, State of California, Richmond, CA 94804, USA. mavis.datechong@doj.ca.gov
authors.forename: Mavis
authors.initials: M
authors.lastname: Date-Chong
authors.forename: Martin R
authors.initials: MR
authors.lastname: Buoncristiani
authors.forename: Margaret
authors.initials: M
authors.lastname: Aceves
authors.forename: Cristián
authors.initials: C
authors.lastname: Orrego
mesh_terms: D002467:Cell Nucleus / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D004272:DNA, Mitochondrial / Q000235:genetics*; D006197:Hair / Q000737:chemistry*; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013499:Surface Properties
chemical_list: D004272:DNA, Mitochondrial; D004247:DNA