The detection of YMDD mutation of hepatitis B virus by allele-specific PCR and a cross-contamination proofed device.
Pak J Pharm Sci
; 28(4): 1311-4, 2015 Jul.
Article
en En
| MEDLINE
| ID: mdl-26142504
ABSTRACT
Lamivudine (LAM) is commonly used to treat Hepatitis B virus (HBV) infection, but its use frequently induces drug resistance. Therefore, rapid and correct detection of drug-resistant HBV is important for effective treatment of HBV infection. In this study, we aimed to develop a novel, simple, and user-friendly method for the detection of LAM resistant HBV. Samples were collected from 60 HBV infected patients for the analysis by allele-specific polymerase chain reaction (AS-PCR), nucleic acid detection strip (NADS) and a cross-contamination proofed device. HBV YMDD mutations were detected by AS-PCR, restriction fragment length polymorphism (RFLP) and DNA sequencing. A 91.7% concordance between all three methods was obtained. Compared to sequencing and RFLP, AS-PCR detected more samples with mutant variants and was more sensitive. This novel method had a detection limit of approximately 103 copies/ml and detected a variant of only 5% of total HBV population. In conclusion, we develop a new assay which could be useful for the detection of HBV LAM resistance, especially in resource-poor settings.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Virus de la Hepatitis B
/
Reacción en Cadena de la Polimerasa
/
Secuencias de Aminoácidos
/
ADN Polimerasa Dirigida por ADN
/
Alelos
/
Mutación
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
Idioma:
En
Revista:
Pak J Pharm Sci
Asunto de la revista:
FARMACIA
/
FARMACOLOGIA
/
QUIMICA
Año:
2015
Tipo del documento:
Article
País de afiliación:
China