Evaluation of different methods in monitoring YMDD motif mutations associated with lamivudine resistance / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 427-430, 2003.
Article
in Chinese
| WPRIM
| ID: wpr-305908
ABSTRACT
<p><b>OBJECTIVES</b>To evaluate the three different methods in monitoring the lamivudine-resistant HBV mutants in lamivudine-treated patients with chronic hepatitis B.</p><p><b>METHODS</b>The sensitivity and specialty of melting curve assay and polymerase chain reaction microplate nucleotide hybridization-enzyme linked immunosorbent assay (PCRmnh-ELISA) were compared with those of mismatch polymerase chain reaction-restriction fragment length polymorphism (mPCR-RFLP) and sequence analysis, through detection of HBV YMDD mutants in 44 serums from chronic hepatitis B patients receiving lamivudine monotherapy at the time of viral breakthrough.</p><p><b>RESULTS</b>mPCR-RFLP assay was more sensitive (10(4) copies/ml) than both PCRmnh-ELISA (10(5) copies/ml) and melting curve assay (10(6) copies/ml). 26 YMDD mutants and 18 wild-types were determined by the means of mPCR-RFLP. Among the 26 mutants, only 16 and 18 mutants were found by melting curve assay and PCRmnh-ELISA, respectively. Whereas, out of the 18 wild-types, 2 and 13 mutants were detected by melting curve assay and PCRmnh-ELISA, respectively. To confirm the different results determined by the three methods in 16 samples, sequence analysis was conducted and showed that the rate of consistency with sequencing was 93.8% by mPCR-RFLP, 43.8% by melting curve, and 18.8% by PCRmnh-ELISA, respectively (chi2=18.7, P<0.01).</p><p><b>CONCLUSIONS</b>The mPCR-RFLP assay is reliable to monitor HBV YMDD mutations. Melting curve assay and PCRmnh-ELISA should be further improved to increase their sensitivity and specialty.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Antiviral Agents
/
Virology
/
Blood
/
Polymorphism, Restriction Fragment Length
/
DNA, Viral
/
Gene Products, pol
/
Hepatitis B virus
/
Polymerase Chain Reaction
/
Sensitivity and Specificity
/
Lamivudine
Type of study:
Diagnostic study
Limits:
Adult
/
Female
/
Humans
/
Male
Language:
Chinese
Journal:
Chinese Journal of Hepatology
Year:
2003
Type:
Article
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