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The detection of YMDD mutation of hepatitis B virus by allele-specific PCR and a cross-contamination proofed device.
Author(s):
1. Wanchuan Zhuang: Department of Hematology, Lianyungang Second People’s Hospital (Lianyungang Hospital affiliated to Bengbu Medical College, Lianyungang, China
2. Ning Sha: Department of Pediatrics, Huai’an Second People’s Hospital Affiliated to Xuzhou Medical College, Huai’an, China
3. Chun Lyu: Department of Hepatology, Huai`an Fourth People`s Hospital, Huai’an, China
4. Jiaxin Li: Department of Pediatrics, Huai`an First People’s Hospital, Nanjing Medical University, Huai’an, China
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.
Page(s): 1311-1314
DOI: DOI not available
Published: Journal: Pakistan Journal of Pharmaceutical Sciences, Volume: 28, Issue: 4, Year: 2015
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