Real-time PCR detection of multiple lamivudine-resistant mutations with displacing probes in a single tube.
J Clin Virol
; 38(4): 334-40, 2007 Apr.
Article
en En
| MEDLINE
| ID: mdl-17329165
ABSTRACT
BACKGROUND:
Detection of lamivudine-resistant hepatitis B virus (HBV) is essential to clinical diagnosis and treatment.OBJECTIVES:
To establish a single tube, real-time PCR assay for simultaneous detection of multiple lamivudine-resistant mutations in serum samples. STUDYDESIGN:
By using four sequence-specific displacing probes labeled with different fluorophores, a single real-time PCR reaction can tell whether a sample contains any of the following HBV variants wild-type, rtM204 mutant; mixtures of wild-type and rtM204 mutant; mixtures of rtM204 and rtL180 mutant; mixtures of wild-type, rtM204 mutant and rtL180 mutant. The assay was evaluated with 50 HBV mutation(s)-containing samples and 36 HBeAg-positive samples.RESULTS:
The results of the real-time PCR assay were consistent with the DNA sequencing, but with much higher sensitivity for detecting a mixture of quasispecies. As few as 10(2)-10(3)copies/ml HBV of all four sequences in pure population and as little as 5% mutant DNA in the presence of wild-type DNA can be detected.CONCLUSIONS:
Application of this high throughput assay into clinical use should enable earlier diagnosis and better treatment of lamivudine-resistant HBV.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Antivirales
/
Virus de la Hepatitis B
/
Reacción en Cadena de la Polimerasa
/
Lamivudine
/
Farmacorresistencia Viral
/
Hepatitis B
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
Límite:
Humans
Idioma:
En
Revista:
J Clin Virol
Asunto de la revista:
VIROLOGIA
Año:
2007
Tipo del documento:
Article
País de afiliación:
China