贵州侗族、苗族和汉族人群乙型肝炎病毒基因型分布

目的　了解贵州侗、苗、汉族HBV感染者的基因型。方法　比较 12 7株各基因型HBV全序列S基因核苷酸序列 ,用DNA分析软件筛选出 3个限制性内切酶。设计 3条HBVS区引物并进行聚合酶链反应扩增 ,产物经MboⅠ、BstNⅠ或BsmAⅠ酶切 ,分析酶切产物电泳图谱 ,建立区分HBV(A～F)基因型的方法。对贵州 16 6份侗、苗、汉族HBV感染者血清进行基因分型 ,5份PCR产物直接测序验证分型方法的准确、可靠。结果　S基因PCR RFLP分型结果准确 ,5份酶切鉴定结果经测序证实。 16 6份标本中 ,B基因型 138份 (83 13% ) ,C型 2 8份 (16 87% ) ,未发现B、C以外的其他基因型。侗族的 4 8例中 ,4 7例为B型 (97 92 % ) ,1例C型 (2 0 8% ) ,苗族的 5 2例中 ,4 9例为B型(94 2 3% ) ,3例C型 (5 77% ) ;6 6例汉族也以B基因为主 (6 3 6 4 % ,4 2 6 6 ) ,但C型有 2 4例 (36 36 % ) ,与侗、苗族HBV感染者相比 ,差异有显著意义 (χ2 =35 0 5 88,P <0 0 0 5 )。结论　贵州地区HBV基因型由B、C 2型构成 ,且以B基因型为主。汉族患者C型较多 ,而侗、苗族患者中B型是占绝对优势的基因型。

Distribution of hepatitis B virus genotype among population of Dong,Miao minority and Han in Guizhou

OBJECTIVE: To study the distribution of hepatitis B virus (HBV) genotype in population of Dong, Miao minority and Han in Guizhou. METHODS: S region nucleotides were compared in 127 strains whole sequence of HBV and three restriction enzymes which can be used for genotyping were found by DNA software analysis system. The partial gene fragment of HBV S region was amplified by nested polymerase chain reaction (nPCR). The products were digested with Mbo I, BstN I or BsmA I and subjected to electrophoresis on agarose gel, respectively. The patterns of restriction fragment length polymorphism (RFLP) were analysed. The genotypes were determined by nPCR-RFLP in 166 asymptomatic HBV carriers (ASC), including 48 Dong minority, 52 Miao minority and 66 Han subjects. Some of the ASC were also analysed by direct sequencing of PCR products. RESULTS: The nPCR-RFLP method was simple and accurate. Of the 166 ASC, 138 (83.13%) were genotype B, and 28 (16.87%) were genotype C. Of the 48 Dong minority subjects, 47 (97.2%) were genotype B and 1 (2.08%) was type C. Of the 52 Miao minority subjects, 49 (94.23%) were genotype B and 3 (5.77%) were genotype C. Of the 66 Han subjects, 42 (63.64%) were genotype B and 24 (36.36%) were genotype C. There was a statistical significance in the distribution of genotype C between Dong, Miao minority and Han (7.85% vs 36.36% P less than 0.005). CONCLUSION: Genotype B and C exist in Guizhou and genotype B is the major genotype. Genotype C is found more frequently in Han than in Dong and Miao minority.