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Infection ; 47(5): 793-803, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30963405

RESUMO

PURPOSE: The frequency of detection of HBV co-infection with multiple HBV genotypes is influenced by the detection method; usually co-infections are detected by multiplex PCR or hybridization assays, and are rarely confirmed by sequencing and conventional cloning. The objective of this study was to confirm by ultra-deep pyrosequencing (UDPS) mixed HBV infections, previously detected by multiplex-nested PCR. METHODS: Sixteen samples from HBV co-infected Sudanese patients detected by multiplex-nested PCR, were amplified targeting the P/S region and sequenced by UDPS. RESULTS: The only genotypes identified using UDPS were D and E, while A, B, C and F genotypes, previously detected by multiplex-nested PCR, were not detected. Specifically, 10 samples were shown to be mono-infected (D or E); in 3 out of 10 mono-infected D patients, a subtype combination was observed: D1 + D7 in 2 cases and D2 + D6 in 1 case. The remaining 6 subjects were D + E co-infected (harboring different mixtures of D subtypes). CONCLUSIONS: Overall, UDPS is more effective than multiplex-nested PCR for identifying multiple HBV genotypes and subtypes infections.


Assuntos
Vírus da Hepatite B/genética , Hepatite B/virologia , Sequenciamento de Nucleotídeos em Larga Escala , Sequência de Aminoácidos , Coinfecção/virologia , DNA Viral/genética , Genótipo , Humanos , Mutação , Filogenia , Análise de Sequência de DNA , Sudão
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