Li Bing, Zhou Bo-Ping, Peng Jin-Fu, Chen Li-Yan, Zhang Wen, Tang Wei, Wang Zhao-Qin, Yin Zhong-Hua, Xu Liu-Mei, Luo Rui-Ling, Li Xiao-He, Liu Sai-Yun
Infection Department, Shenzhen Disanrenmin Hospital of Guangdong Medical College, Shenzhen 518020, China
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Aug;23(4):309-12.
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.
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.
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.
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]
本研究旨在构建一种可用于临床检测乙型肝炎病毒拉米夫定耐药突变体及基本核心启动子/前C区突变体的基因芯片方法,将该方法与DNA测序法进行比较,以研究此基因芯片的特性(灵敏度、特异性、稳定性及临床实用性),并将其应用于临床。
该基因芯片检测方法可检测乙肝病毒的DNA及8个突变位点,包括3个拉米夫定耐药突变位点(DNA聚合酶基因第180、204、207位点),5个HBeAg逃逸相关突变位点(BCP/前C区的nt1896、1899、1862、1764、1762位点)。基因芯片法检测结果通过DNA测序进行验证。
在检测乙肝病毒DNA时,基因芯片结果与DNA测序结果100%相符。在检测乙肝病毒突变体时,两种方法对32份样本中的256个位点检测结果有251个位点相同,仅5个位点不完全匹配(P>0.05)。在这5个位点中,基因芯片法检测为多重感染结果,而测序法检测为单一感染结果。
这些结果表明基因芯片法与DNA测序法阳性率相同,特异性基本一致,且在检测多重感染乙肝病毒株方面优于DNA测序法。[关键词]