Li Xiaodong, Jiang Lihong, Li Fan, Liu Yan, Dai Jiuzeng, Zhao Pan, Qin Yaqun, Li Jin, Xu Dongping
Medical school of Chinese PLA, Beijing 100853, China.
Zhonghua Gan Zang Bing Za Zhi. 2015 Jan;23(1):23-7. doi: 10.3760/cma.j.issn.1007-3418.2015.01.007.
To determine the mutational profile and clinical implications of the viral reverse-transcriptase (rt)A 181T mutation in hepatitis B virus (HBV) through population-based analysis of clinical samples.
Serum samples from 3, 013 patients who visited The 302 Hospital (Beijing, China) were investigated.HBV DNA was extracted and HBV mutations and genotypes were determined by direct sequencing.Recombinant plasmids harboring the rtA181T/sW172* mutant or wild type sequence were constructed and transfected into the HepG2 cell line. The levels of HBsAg in culture supernatants were compared and statistically analyzed.
The incidence of rtA181T across the study population was 4.1% (165/3, 013), and most of the rtAl 81T-positive patients had received adefovir and/or lamivudine.Forty percent (66/165) of the rtA 181T cases were single mutants and treatment responsive, 46.1% (76/165) included the adefovir-resistant mutation rtA 181 V/N236T, 12.1% (20/165) included the lamivudine-resistant mutation rtM204V/rtM2041, and 1.8% (3/165) included multidrug-resistant mutations.Interestingly, 73.9% (122/165) of the rtA181T-positive samples were detected with co-existing wild-type nucleotides at the site. The rates of HBV/C to HBV/B were 92.1% to 7.9% in the rtA181T-positive patients, but 82.1% to 17.9% in the rtA181T-negative paticnts (P less than 0.01).Almost all (98.2%; 129/165) of the rtA181T led to sW172*, while only 1.8% of the rtA181T (3/165) led to sW172L or sW172S.HBsAg secretion in vitro was reduced from the rtA181T/ sW172* strain, but there was no significant difference observed in the average serum HBsAg and HBV DNA levels of patients who carried or did not carry the mutant.
The HBV rtA181T mutation is closely associated with adefovir and lamivudine exposure.rtA181T may led to sW172*, culminating in suppression of HBsAg secretion.However, co-existence of the mutant with wild-type sequences was common among our patient population, suggesting that the mutation had little impact on serum HBsAg and HBV DNA levels across the clinical study population.
通过对临床样本进行基于人群的分析，确定乙型肝炎病毒（HBV）中病毒逆转录酶（rt）A181T突变的突变谱及其临床意义。
对就诊于中国人民解放军第三〇二医院（北京）的3013例患者的血清样本进行研究。提取HBV DNA，通过直接测序确定HBV突变和基因型。构建携带rtA181T/sW172*突变体或野生型序列的重组质粒，并转染至HepG2细胞系。比较并统计分析培养上清液中HBsAg的水平。
在整个研究人群中，rtA181T的发生率为4.1%（165/3013），大多数rtA181T阳性患者接受过阿德福韦和/或拉米夫定治疗。40%（66/165）的rtA181T病例为单突变且对治疗有反应，46.1%（76/165）包含阿德福韦耐药突变rtA181V/N236T，12.1%（20/165）包含拉米夫定耐药突变rtM204V/rtM204I，1.8%（3/165）包含多药耐药突变。有趣地是，73.9%（122/165）的rtA181T阳性样本在该位点检测到野生型核苷酸共存。rtA181T阳性患者中HBV/C与HBV/B的比例为92.1%至7.9%，而rtA1S1T阴性患者中为82.1%至17.9%（P<0.01）。几乎所有（98.2%；129/165）的rtA181T导致sW172*，而只有1.8%（3/165）的rtA181T导致sW172L或sW172S。rtA181T/sW172*毒株体外HBsAg分泌减少，但携带或未携带该突变的患者血清HBsAg和HBV DNA平均水平无显著差异。
HBV rtA181T突变与阿德福韦和拉米夫定暴露密切相关。rtA181T可能导致sW17S*，最终抑制HBsAg分泌。然而，在我们的患者群体中，突变体与野生型序列共存很常见，这表明该突变对整个临床研究人群的血清HBsAg和HBV DNA水平影响较小。
