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Titolo:
Validation of short tandem repeats (STRs) for forensic usage: Performance testing of fluorescent multiplex STR systems and analysis of authentic and simulated forensic samples
Autore:
Moretti, TR; Baumstark, AL; Defenbaugh, DA; Keys, KM; Smerick, JB; Budowle, B;
Indirizzi:
Fed Bur Invest Acad, Forens Sci Res Unit, Sci Anal Sect, FBI Lab, Quantico, VA 22135 USA Fed Bur Invest Acad Quantico VA USA 22135 FBI Lab, Quantico, VA 22135 USA Lab Div, DNA Anal Unit 1, Sci Anal Sect, Washington, DC USA Lab Div Washington DC USA Anal Unit 1, Sci Anal Sect, Washington, DC USA
Titolo Testata:
JOURNAL OF FORENSIC SCIENCES
fascicolo: 3, volume: 46, anno: 2001,
pagine: 647 - 660
SICI:
0022-1198(200105)46:3<647:VOSTR(>2.0.ZU;2-6
Fonte:
ISI
Lingua:
ENG
Soggetto:
DEOXYRIBONUCLEIC-ACID DNA; POLYMERASE CHAIN-REACTION; PCR AMPLIFICATION; CASEWORK; LOCI; IDENTIFICATION; POLYMORPHISMS; ENVIRONMENT; EXTRACTION; SUBSTRATE;
Keywords:
forensic science; short tandem repeats; core CODIS loci; CSF1PO; FGA; TH01; TPOX; vWA; D3S1358; D5S818; D7S820; D8S1179; D13S317; D16S539; D18S51; D21S11; polymerase chain reaction; multiplex amplification; fluorescence; DNA typing; validation;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Citazioni:
41
Recensione:
Indirizzi per estratti:
Indirizzo: Moretti, TR Fed Bur Invest Acad, Forens Sci Res Unit, Sci Anal Sect, FBI Lab, Quantico, VA 22135 USA Fed Bur Invest Acad Quantico VA USA 22135 ntico, VA 22135 USA
Citazione:
T.R. Moretti et al., "Validation of short tandem repeats (STRs) for forensic usage: Performance testing of fluorescent multiplex STR systems and analysis of authentic and simulated forensic samples", J FOREN SCI, 46(3), 2001, pp. 647-660

Abstract

The amplification and typing conditions for the 13 core CODIS loci and their forensic applicability were evaluated. These loci are CSF1PO, FGA, TH01,TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, andD21S11. Results were obtained using the multiplex STR systems AmpFlSTR((R)) Profiler Plus (TM) and AmpFlSTR COfiler (TM) (Applied Biosystems, Foster City, CA), GenePrint (TM) PowerPlex (TM) (Promega Corporation, Madison, WI), and subsets of these kits. For detection of fluorescently labeled amplified products, the ABI Prism((R)) 310 Genetic Analyzer, the ABI Prism 377 DNASequencer, the FMBIO(R) II Fluorescent Imaging Device, and the FluorImager(TM) were utilized. The following studies were conducted: (a) evaluation of PCR parameter ranges required for adequate performance in multiplex amplification of STR loci, (b) determination of the sensitivity of detection of the systems, (c) characterization of non-allelic PCR products, (d) evaluation of heterozygous peak intensities, (e) determination of the relative level of stutter per locus, (f) determination of stochastic PCR thresholds, (g)analysis of previously typed case samples, environmentally insulted samples, and body fluid samples deposited on various substrates. and (h) detection of components of mixed DNA samples. The data demonstrate that the commercially available multiplex kits can be used to amplify and type STR loci successfully from DNA derived from human biological specimens. There was no evidence of false positive or false negative results and no substantial evidence of preferential amplification within a locus. Although at times generalbalance among loci labeled with the same fluorophore was not observed, theresults obtained were still valid and robust. Suggested criteria are provided for determining whether a sample is derived from a single source or from more than one contributor. These criteria entail the following: (a) the number of peaks at a locus, (b) the relative height of stutter products, and(c) peak height ratios. Stochastic threshold levels and the efficiency of non-templated nucleotide addition should be considered when evaluating the presence of mixtures or low quantity DNA samples. Guidelines, not standards, for interpretation should be developed to interpret STR profiles in cases, because there will be instances in which the standards may not apply. These instances include (a) a primer binding site variant for one allele at a given locus, (b) unusually high stutter product, (c)gene duplication, and (d) translocation.

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Documento generato il 30/11/20 alle ore 19:44:35