Tian Guo-bao, Zeng Zheng, Huang Zhong-hong, Lu Hai-ying, Yu Min, Gong Wei-bo, Wang Dong, Si Chong-wen
Department of Infectious Diseases, Peking University First Hospital, Beijing 100034, China
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2011 Feb;25(1):23-5.
To investigate the variants and quasispecies of reverse transcriptase region in polymerase gene of hepatitis B virus (HBV) during lamivudine treatment and their relationship with genotypes and viral loads.
HBV DNA of 117 chronic hepatitis B patients treated with lamivudine were amplified by using PCR. The PCR products including the YMDD motif were sequenced by DNA sequencer, of which, HBV DNA viral loads of 99 patients were determined by real-time PCR and 64 samples were sequenced by Pyrosequencing.
In HBV YMDD variant group and no variant group, the HBV genotypes were 79.6% and 86.7% of type C, 18.5% and 12.7% of type B, 1.9% of A/B recombinant type and 2.6% of type D, respectively. The viral loads (log 10) were 6.5699 and 6.6165, respectively. There was no significant difference in HBV genotypes and viral loads between these two groups. The rtL180M variant was found in association with the rtM204I/V variant, HBV variants and wild-type in YMDD motif all existed together in these two groups.
HBV variants (quasispecies) in YMDD motif could be quantified by pyrosequencing, which would be a feasible measure during nucleoside or nucleotide analogue therapy against chronic HBV infection.
研究拉米夫定治疗期间乙型肝炎病毒(HBV)聚合酶基因逆转录酶区的变异及准种情况,以及它们与基因型和病毒载量的关系。
采用聚合酶链反应(PCR)扩增117例接受拉米夫定治疗的慢性乙型肝炎患者的HBV DNA。对包括YMDD基序的PCR产物进行DNA测序,其中99例患者的HBV DNA病毒载量通过实时PCR测定,64份样本通过焦磷酸测序进行测序。
在HBV YMDD变异组和无变异组中,HBV基因型C型分别占79.6%和86.7%,B型分别占18.5%和12.7%,A/B重组型占1.9%,D型占2.6%。病毒载量(log10)分别为6.5699和6.6165。两组间HBV基因型和病毒载量无显著差异。发现rtL180M变异与rtM204I/V变异相关,两组中YMDD基序中的HBV变异体和野生型均同时存在。
焦磷酸测序可对YMDD基序中的HBV变异体(准种)进行定量分析,这在核苷或核苷酸类似物治疗慢性HBV感染过程中是一种可行的方法。