Multiplex amplification and typing procedure for the loci D1S80 and amelogenin.
ABSTRACT A method has been developed that enables multiplex amplification and simultaneous typing of the loci D1S80 and amelogenin using discontinuous polyacrylamide gel electrophoresis and silver staining. The protocol is sensitive, simple, rapid, and relatively inexpensive. The results of the multiplex analysis of the D1S80 and amelogenin loci were comparable to those obtained when each locus was analyzed individually. A small validation study was undertaken to evaluate the forensic applicability of this multiplex system. The data demonstrate that DNA exposed to a variety of environmental insults yields reliable multiplex typing results.
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ABSTRACT: Surgical cure of glioblastomas is virtually impossible and their clinical course is mainly determined by the biologic behavior of the tumor cells and their response to radiation and chemotherapy. We investigated whether response to temozolomide (TMZ) chemotherapy differs in subsets of malignant glioblastomas defined by genetic lesions. Eighty patients with newly diagnosed glioblastoma were analyzed with comparative genomic hybridization and loss of heterozygosity. All patients underwent radical resection. Fifty patients received TMZ after radiotherapy (TMZ group) and 30 patients received radiotherapy alone (RT group). The most common aberrations detected were gains of parts of chromosome 7 and losses of 10q, 9p, or 13q. The spectrum of genetic aberrations did not differ between the TMZ and RT groups. Patients treated with TMZ showed significantly better survival than patients treated with radiotherapy alone (19.5 vs 9.3 months). Genomic deletions on chromosomes 9 and 10 are typical for glioblastoma and associated with poor prognosis. However, patients with these aberrations benefited significantly from TMZ in univariate analysis. In multivariate analysis, this effect was pronounced for 9p deletion and for elderly patients with 10q deletions, respectively. This study demonstrates that molecular genetic and cytogenetic analyses potentially predict responses to chemotherapy in patients with newly diagnosed glioblastomas.Neoplasia 11/2005; 7(10):883-93. · 5.47 Impact Factor
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ABSTRACT: Allele frequencies for the seven PCR-based loci (LDLR, GYPA, HBGG, D7S8, Gc, HLA-DQAI, and D1S80) were determined in a Black African population from Zimbabwe. All loci are highly polymorphic and meet Hardy-Weinberg expectations. An interclass correlation analysis detected only two significant departures from independence out of 21 pair-wise comparisons of the 7 loci. The Black African allele frequency data are similar to African American data at four of the seven PCR-based loci.Deutsche Zeitschrift für die Gesamte Gerichtliche Medizin 02/2000; 113(5):300-1. · 2.69 Impact Factor
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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, and D21S11. Results were obtained using the multiplex STR systems AmpFlSTR Profiler Plus and AmpFlSTR COfiler (Applied Biosystems, Foster City, CA), GenePrint PowerPlex (Promega Corporation, Madison, WI), and subsets of these kits. For detection of fluorescently labeled amplified products, the ABI Prism 310 Genetic Analyzer, the ABI Prism 377 DNA Sequencer, the FMBIO II Fluorescent Imaging Device, and the Fluorlmager 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 general balance among loci labeled with the same fluorophore was not observed, the results 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.Journal of Forensic Sciences 06/2001; 46(3):647-60. · 1.24 Impact Factor