[Construction and application of a genechip method for detection of hepatitis B virus lamivudine-resistant mutants and basal core promotor/Pre-C mutants].
OBJECTIVE
The objective of this research is to construct a clinic-usable genechip method for detection of hepatitis B virus lamivudine-resistant mutants and basal core promotor/Pre-C mutants, compare this method with DNA sequencing to investigate this genechip's character (sensity, specificity, stability and practicability in clinic) and apply it in clinic.


METHODS
This genechip detection method can detect the DNA and 8 mutative site of HBV, include 3 lamivudine-resistant mutation site(No. 180, 204, 207 site in DNA polymerase gene), 5 HBeAg escape-related mutation site (nt 1896, 1899, 1862, 1764,1762 site in BCP/Pre-C region).The results of genechip method was verified by DNA sequencing.


RESULTS
In detecting HBV DNA, the results of genechip were agree with 100% of the results of DNA sequencing. In detecting HBV mutants, 251 sites (in 32 samples, 256 sites) showed the same results using both methods, and only 5 sites were not completely match (P > 0.05). In these 5 sites, genechip methods got multi-infection results, but sequencing got single-infection results.


CONCLUSIONS
These results suggest that genechip method has the same positive rate and almost these same specificity with DNA sequencing method, and is better than DNA sequencing method in detecting multi-infected HBV strains. [Key words]
Li
Bing
B
Infection Department, Shenzhen Disanrenmin Hospital of Guangdong Medical College, Shenzhen 518020, China
Zhou
Bo-Ping
BP
Peng
Jin-Fu
JF
Chen
Li-Yan
LY
Zhang
Wen
W
Tang
Wei
W
Wang
Zhao-Qin
ZQ
Yin
Zhong-Hua
ZH
Xu
Liu-Mei
LM
Luo
Rui-Ling
RL
Li
Xiao-He
XH
Liu
Sai-Yun
SY
D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D024882:Drug Resistance, Viral; D006509:Hepatitis B / Q000188:drug therapy / Q000821:virology*; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D019259:Lamivudine / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011401:Promoter Regions, Genetic
D000998:Antiviral Agents; D006512:Hepatitis B Core Antigens; D019259:Lamivudine