Detection of drug resistance mutation in HBV using matrix assisted laser desorption/ionization time-of-flight mass spectrometry
10.3760/cma.j.issn.1009-9158.2011.03.006
- VernacularTitle:飞行时间质谱检测HBV耐药位点变异及其影响因素探讨
- Author:
Yingxin HAN
;
Yanfang GUAN
;
Jingjing LI
;
Yinhong ZHAO
;
Huanming YANG
;
Jian HUANG
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus;
Drug resistance,viral;
Variation (genetics);
Spectrometry,mass,electrosprayionization
- From:
Chinese Journal of Laboratory Medicine
2011;34(3):213-217
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a rapid method for detection of drug-resistance mutation in HBV, based on PCR-MALDI-TOF MS, and to explore the influential factors on this method. Methods One hundred blood serum samples, which were collected from chronic HBV patients with single drug-resistance or multiple drug-resistance of Lamivudin, Adefovi, Entecavir and Telbivudine, and 10 kinds of mutant HBV plasmids were analyzed using PCR-MALDI-TOF MS and confirmed by PCR-based sequencing. Results Of 100 samples detected, thirty-one samples were positive for drug-resistance and 69 samples were negative. The PCR-MALDI-TOF MS results of 94 samples were completely consistent with PCR-based sequencing. Six samples were inconsistent , of which three samples were positive by the two methods, but more mutation loci were detected by PCR-MALDI-TOF MS than sequencing. The consistent rate of two methods was 94%,detection sensitivity was up to 100 copies/μl, and the cut off value of detectable mutation level was 5%.Conclusion PCR-MALDI-TOF MS could be used for rapid and simple analysis of the drug resistance for the clinical application with features of high sensitivity and accuracy, high throughput and automation.
