摘要
目的调查昆明、洛阳两地献血者人群HBV携带者HBV基因型的分布情况。方法采用多对型特异性引物,以套式PCR的方法对两地367份献血者HBsAg筛查阳性样本分型,根据HBV前s1基因和s基因区域内的保守序列设计出10条引物(2条外引物、8条内引物),并将其中8条内引物分为Ⅰ、Ⅱ组,分别扩增A、B、C和D、E、F型HBV基因片段;将第2轮PCR产物做琼脂糖电泳,根据PCR产物片断大小判定基因型;分型结果以s基因直接测序法验证。结果昆明279份样本中成功分型214例,分型成功率76.7%,其中B型69例(32.2%),C型103例(48.1%),B、C混合型41例(19.2%);D型1例(0.5%);洛阳献血者的88份样本中成功分型72例,分型成功率81.8%,其中B型10例(13.9%),C型46例(63.9%),B、C混合型16例(22.2%)。经s基因测序法验证,该分型方法准确可靠。结论昆明、洛阳两地献血者人群中HBV携带者HBV基因型均以C型为主,并有一定比例的B型和B、C混合型,仅在昆明献血者中发现1例D型,未见A、E、F型。
Objective To investigate the distribution of hepatitis B virus （HBV） genotypes among blood donors in Kunming and Luoyang via nested PCR with multiplex pairs of genotype-specific primers. Methods A total of 10 primers, including 2 outer and 8 inner primers, were designed on the basis of the conserved nucleotide sequences in regions of the Pre-S1 through S genes, and the 8 genotype-specific inner primers were divided into 2 groups,mix A and B to amplify HBV of genotype A, B, C or D, E, F, respectively. Electrophoresis of the two groups＇PCR products of the second-round in agarose gel was employed to identify the genotypes. S gene sequencing was used to assess the accuracy. Results Among the 88 blood samples of Luoyang donors ,72 were genotyped suceessfully, among which 10（ 13.9% ） were genotype B , 46（63. 9% ） were genotype C, 16 （22.2%） were mixed genotype （ B ＋ C）. Among the 279 blood samples of Kunming donors, 214 were genotyped suceessfully, among which 69 （ 32.2% ） were genotype B, 103 （48.1% ） were genotype C, 41 （ 19.2% ） were mixed genotype （B ＋ C）, 1 （0.5 % ）were genotype D. The results, confirmed by direct sequencing of S gene, were accurate and reliable. Conclusion Genotype C is predominant in both Kunming and Luoyang donors with hepatitis B, while genotype B and mixed genotype （B ＋ C） also exist. One ease of genotype D was detected among Kunming donors, and no A, E and F genotypes were found.
出处
《中国输血杂志》
CAS
CSCD
北大核心
2009年第7期552-555,共4页
Chinese Journal of Blood Transfusion
基金
美国NHLBI项目REDS-Ⅱ International基金(NO.8067-5003)部分赞助
