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Brazilian Journal of Infectious Diseases

Print version ISSN 1413-8670

Abstract

WANG, Yong-Zhong et al. Detection of hepatitis B virus A1762T/G1764A mutant by amplification refractory mutation system. Braz J Infect Dis [online]. 2014, vol.18, n.3, pp.261-265. ISSN 1413-8670.  https://doi.org/10.1016/j.bjid.2013.09.005.

OBJECTIVE:

To study the role of hepatitis B virus with A1762T/G1764A double mutation in liver cirrhosis and hepatocellular carcinoma, and create a sensitive, fast, accurate assay for detection of A1762T/G1764A double mutation.

METHODS:

We developed an accurate and fast real-time amplification refractory mutation system to detect A1762T/G1764A double mutation. Cloned hepatitis B virus genome was used as a control. Assay sensitivity was determined by serial dilution and mixed template experiments. Specificity was determined by cross experiments with wild and mutant hepatitis B virus. Fifty clinical samples were tested by the real-time amplification refractory mutation system and the results were compared with sequencing.

RESULTS:

The real-time amplification refractory mutation system had a sensitivity of 100 copies of virus with these mutations, and 0.1% weak population virus with double mutation could be found in mixtures. A total of 50 randomly collected clinical samples were detected by real-time amplification refractory mutation system, and the results were consistent with those by DNA sequencing. Hepatitis B virus genotype C was more prevalent in 39 of 50 samples than genotype B (11 samples), and about 75% of genotype C carried a double mutation compared to 45% of genotype B. However, the percentage of A1762T/G1764A double mutation in hepatitis B e antigen-negative (58.3%) samples was almost the same as in hepatitis B e antigen-positive (61%) samples.

CONCLUSION:

The real-time amplification refractory mutation system is sensitive and specific for detection of hepatitis B virus double mutation.

Keywords : Hepatitis B virus; Basal core promoter mutations; Amplification refractory mutation system; PCR.

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