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J Med Virol ; 83(6): 1042-7, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21503918

RESUMO

In order to investigate the influence of DNA extraction on two PCR-based HPV genotyping tests (Linear Array, Roche and INNO-LiPA Extra, Innogenetics), three different procedures were used to purify DNA from 28 cervico-vaginal samples tested previously by the Hybrid Capture 2: the AmpliLute Liquid Media Extraction kit (Roche), the QIAamp DNA Blood mini kit (QIAGEN), and the NucliSENS EasyMAG automated platform (bioMérieux). All HC2-positive samples were found positive by both assays, independently of the extract used. Type-specific concordance (i.e., identical HPV type-specific profile in all the extracts of the same sample) was observed in 55% and 75% of the cases testing samples by the Linear Array and the INNO-LiPA, respectively. Using the DNA extracted with the two manual methods the results were concordant in 75% of the cases both for the Linear Array and the INNO-LiPA. When comparing the Linear Array results obtained on either of the two manual extracts with those obtained following automated extraction, 65% of the samples showed type-specific concordance in both cases. The INNO-LiPA results were concordant in 80% of the cases comparing the AmpliLute versus the automated extract, while concordant results were observed in 90% of the cases when comparing the QIAGEN versus the automated extract. In conclusion, the Linear Array and INNO-LiPA results are affected by the method of DNA extraction. Consequently, different HPV type-specific profiles may be observed using different extracts of the same sample. The use of consistent protocols for DNA purification is a priority to guarantee intra-assay reproducibility over time.


Assuntos
Alphapapillomavirus/isolamento & purificação , DNA Viral/isolamento & purificação , Técnicas de Genotipagem/métodos , Infecções por Papillomavirus/virologia , Infecções Tumorais por Vírus/virologia , Alphapapillomavirus/classificação , Alphapapillomavirus/genética , Colo do Útero/virologia , DNA Viral/genética , Feminino , Genótipo , Técnicas de Genotipagem/normas , Humanos , Infecções por Papillomavirus/genética , Reação em Cadeia da Polimerase/métodos , Gravidez , Complicações Infecciosas na Gravidez/genética , Complicações Infecciosas na Gravidez/virologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Infecções Tumorais por Vírus/genética , Vagina/virologia , Esfregaço Vaginal/métodos
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