[High throughput detection of drug-resistance gene mutations in HBV using MALDI-TOF mass spectrometry].
OBJECTIVE
To develop a high-throughput clinical method on drug-resistance gene mutations of HBV using MALDI-TOF-MS.


METHODS
Using MassArray Assay Design software designed the iPLEX primers and followed the iPLEX instruction for amplification, SAP reaction, primer extenction, desalination, dispensing, MALDI-TOF-MS screening and data analysis of the gene mutation locus. 138 serum samples of chronic HBV patients with single drug-resistance or multiple drug-resistance on Lamivudin, adefovi, Entecavir were detected.


RESULTS
The HBV gene mutation platform was successfully developed and applied on the high-throughput dectection of clinical serum samples. It was also a high throughput assay which could be used to detect for more than 138 samples once. The MALDI-TOF-MS technology and the DNA sequencing simultaneously examine 33 samples, in which result of 10 sample is inconsistent, the including 2 samples by MALDI-TOF-MS technology has not tested, 1 sample has 2 inconsistent mutations.


CONCLUSIONS
Detection of HBV gene mutations using MALDI-TOF-MS is highly-sensitive, highly-accurate, high-throughput, fast achieved and suitable to use in the diagnosis and monitoring of HBV.
Zhou
Bo-ping
BP
Shenzhen Donghu Hospital, Shenzhen 548020, China.
Zhang
Hong-mei
HM
Li
Xiao-he
XH
Yuan
Jing
J
Li
Wei
W
Xu
Liu-mei
LM
Wang
Huo-sheng
HS
Chen
Xin-chun
XC
Ding
Chun-ming
CM
D004279:DNA, Viral / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics*; D024882:Drug Resistance, Viral / Q000187:drug effects* / Q000235:genetics*; D006515:Hepatitis B virus / Q000187:drug effects*; D013058:Mass Spectrometry; D020641:Polymorphism, Single Nucleotide
D004279:DNA, Viral