摘要
目的了解兰州地区无偿献血者乙肝病毒的感染情况,分析其S区基因的特征,探明本地区实行核酸筛查的安全性和可靠性,为HBV感染的监测策略提供重要依据,持续改进血液筛查方法,降低经输血传播HBV的风险。方法对2015年1月—2016年12月收集到献血者标本采用ELISA进行HBsAg筛查,将HBsAg(+)标本进行中和试验,对中和试验阳性和HBsAg(-)/HBV DNA(+)标本进行核酸提取,然后按照HBV/HCV/HIV-1 3项目分项做核酸病毒检测,再对HBV核酸检测阳性标本做S区基因分型,运用Mega 5.0将HBV S基因分型的结果做系统进化树,并分析其S区的氨基酸序列的变异。结果本次共筛查108 244份血液标本,HBsAg阳性率为0.34%;NAT检测108 045份,共有HBV DNA反应性标本32份,流行率为1∶3 376 (32/108 045);中和试验有63份标本检测,其中40份为阳性,中和阳性率为63.5%。40份中和试验为阳性的标本中,26份标本成功测通S区基因,其分型结果为C型26例;32份HBV DNA反应性标本中,11份成功测通S区基因序列,其分型结果为C型9例、D型2例。分析ELISA与NAT之间在S区的氨基酸序列的差异性,发现主要的突变位点是sV126 L/T/I,sT148L/V,sC149F/V,sA159G/D,sR160D/K和sW163/G/R,而sV168A/K位点可能是本地区的潜在突变位点。结论兰州地区献血者HBV S区流行株C和D,以C基因型为主,其氨基酸序列存在多个位点的变异,这些变异受基因型的影响,这些变异序列为血液筛查策略和筛查试剂的选择提供了依据。
Objective To understand the infection status of hepatitis B virus(HBV) in voluntary blood donors in Lanzhou area of Northwest China, and characterize HBV S gene mutations, further to verify the safety and reliability of nucleic acid screening in this area so as to provide references for the monitoring strategy of HBV infection and continuous improvement of screening methodology.Methods HBsAg was screened in the blood samples collected from January 2015 to December 2016 using ELISA. Neutralization test was carried out on HBsAg positive samples.Nucleic acid extraction was conducted on positive samples in neutralization test, as well as samples with HBsAg negative and HBV DNA positive, and then nucleic acid virus test was employed to detect HBV,HCV, and HIV-1 items in these samples. The positive samples in HBV nucleic acid test were amplified for sequence analysis in the HBV S genetic region, and its sequence were analyzed by phylogenetic tree using Mega 5.0, and the variation in amino acid sequence of HBV S region was also analyzed. Results Of the 108 244 blood samples, 0.34% of them were screened as HBsAg positive. Of the 108 045 NAT samples, 32 HBV-DNA reactive samples were found, accounting for 1∶3 376(32/108 045) of the NAT samples. 63 samples underwent the neutralization test with 40 positive samples at a positive rate of 63.5%. Of the 40 neutralization positive samples, 26 had successful amplification of the S genetic region, and all were Genotype C. Of the 32 HBV DNA reactive samples, 11 had successful amplification of the S genetic region, and 9 were Genotype C and 2 were Genotype D.When the amino acid sequence in S region was compared between ELISA and NAT, such main mutation loci as sV126 L/T/I, sT148 L/V, sC149 F/V, sA159 G/D, sR160 D/K, sW163/G/R, and sV168 A/K were found, which may be the potential mutation loci in this region. Conclusion Strains C and D in HBV S region are prevalent in blood donors in Lanzhou, Genotype C,however, is predominant. Multiple HBV genetic variations were observed in the a
作者
潘登
张支凤
吴康乐
秦龑丽
杨树龙
蒋淼
方琴
何爽
PAN Deng;ZHANG Zhifeng;WU Kangle;QIN Yanli;YANG Shulong;JIANG Miao;FANG Qin;HE Shuang(Blood Center of Gansu Province,Lanzhou 730046;Shanghai Kehua Bio-engineering Co.,LTD.)
出处
《中国输血杂志》
CAS
2019年第2期137-141,共5页
Chinese Journal of Blood Transfusion
基金
甘肃省卫生行业科研计划项目(GSWSKY2016-42)
关键词
兰州
献血者
乙型肝炎病毒
基因型
S区突变
Lanzhou
blood donors
hepatitis B virus
genotype
S region mutation
