Detection of hepatitis B virus A1762T/G1764A mutant by amplification refractory mutation system
Braz. j. infect. dis
; 18(3): 261-265, May-June/2014. tab, graf
Article
in En
| LILACS
| ID: lil-712963
Responsible library:
BR1.1
ABSTRACT
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. .Key words
Full text:
1
Index:
LILACS
Main subject:
DNA, Viral
/
Hepatitis B virus
/
Carcinoma, Hepatocellular
/
Liver Cirrhosis
/
Liver Neoplasms
/
Mutation
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
En
Journal:
Braz. j. infect. dis
Journal subject:
DOENCAS TRANSMISSIVEIS
Year:
2014
Type:
Article