Clinical Chemistry
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Clinical Chemistry 45: 178-183, 1999;
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(Clinical Chemistry. 1999;45:178-183.)
© 1999 American Association for Clinical Chemistry, Inc.


Articles

Efficiency of Two Different Nine-Loci Short Tandem Repeat Systems for DNA Typing Purposes

Lucia Sacchetti1, Giuseppe Calcagno1, Iolanda Coto1, Nadia Tinto2, Emilia Vuttariello2 and Francesco Salvatore1,a

1 Dipartimento di Biochimica e Biotecnologie Mediche, Università degli Studi di Napoli "Federico II", 80131 Napoli, Italy.

2 CEINGE-Biotecnologie Avanzate; via S. Pansini 5, 80131 Napoli, Italy.
a Address correspondence to this author at: Dipartimento di Biochimica e Biotecnologie Mediche, Facoltà di Medicina e Chirurgia, Università di Napoli "Federico II", via S. Pansini 5, 80131 Napoli Italy. Fax 39-81-7463650; e-mail salvator{at}unina.it.

Background: Genotyping based on short tandem repeat (STR) regions is widely used in human identification and parentage testing, in gene mapping studies, and as an approach to studies on the etiopathogenesis and diagnosis of hereditary diseases. We wished to study a new analytical approach that uses capillary electrophoresis and multicolor fluorescence in place of slab gel electrophoresis. Methods: We evaluated the efficiency for parentage and forensic purposes of the AmpFLSTR Profiler PlusTM typing kit that is used with the ABI Prism 310 Genetic Analyzer (System-2 STR), and that of a widely used panel of nine STRs analyzed with conventional slab-gel electrophoresis followed by radioactive detection (System-1 STR). System-2 STR, based on automated capillary electrophoresis and automated sizing of the alleles by Genotyper 2.0 software, was used to determine the allele frequency of the nine loci in 157 Caucasian subjects from southern Italy. On the basis of the data obtained, we submitted 40 trios to parentage testing. Results: A higher median probability of paternity attribution and power of exclusion were obtained with System-2 STR vs System-1 STR: respectively, 99.99% and 99.95% (P <0.05) for attribution; and five and four excluding loci (P <0.05) for exclusion. The most informative and highly discriminating loci were D18S51, D21S11, and FGA. The combined probability of matching-by-chance for all nine STRs was 1.36 x 10-12 for System-2 compared with 1.11 x 10-7 obtained with the other system. The internal standard and allelic ladder of the System-2 STR facilitated accurate and precise genotyping; furthermore, System-2 STR and was faster than the conventional System-1 STR. Conclusions: The System-2 STR allows rapid testing with higher probabilities of attribution and a higher power of exclusion than with the comparison method with slab-gel electrophoresis.




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