Ding Jing-juan, Liu Yue-hui, Wang Mei
Department of Infectious Diseases, Affiliated Hospital of Guiyang Medical College, Guiyang 550004, China.
Zhonghua Gan Zang Bing Za Zhi. 2007 Oct;15(10):725-8.
To investigate hepatitis B virus (HBV) subtypes in patients chronically infected with genotype B or C of hepatitis B virus in Guizhou and to study the relationship between the subtypes and the progression of their liver diseases.
Using PCR, 309 bp gene fragments in the HBV p region were amplified. The products of PCR were digested by VspI, NciI, BstEII and subjected to agarose gel electrophoresis. The subtypes of C1 and C2 were detected by restriction fragment length polymorphism (RFLP). B subtype was determined by direct sequencing of PCR product. One hundred seventy-eight patients with genotype B or C HBV infection in Guizhou, including 50 asymptomatic carriers (ASC), 100 chronic hepatitis (CHB), 14 liver cirrhosis (LC), and 14 hepatocellular carcinoma (HCC) patients were examined. The relationship between HBV C subtypes and the progression of their liver diseases was studied by analyzing the HBeAg positivity, HBV DNA loads and ALT levels of the patients.
Of 84 patients with HBV genotype C, 27 (32.14%) and 56 (66.67%) were subtype C1 and C2, respectively. In 94 genotype B, 93 (98.94%) were subtype Ba and only one was subtype Bj. Subtype C1 showed a trend of gradual decrease from ASC and CHB to LC/HCC groups. In contrast, subtype C2 showed a gradual increase (trend) in the same order. The HBeAg positivity was significantly lower in subtype C1 than that in subtype C2. The ALT levels and HBV DNA loads were higher in patients with subtype C2 than those in subtype C1, however no statistical significance was found in these primes (t=0.95, 0.79).
Subtype Ba is major and subtype C2 is more common in Guizhou. The distribution of subtype C1 and C2 are different in various stages of liver disease. The PCR-RFLP method is simple and accurate and can be used in a large-scale survey.
研究贵州慢性感染乙型肝炎病毒B或C基因型患者的乙肝病毒(HBV)亚型,并探讨亚型与肝脏疾病进展之间的关系。
采用聚合酶链反应(PCR)扩增HBV p区309 bp基因片段。PCR产物经VspI、NciI、BstEII酶切后进行琼脂糖凝胶电泳。通过限制性片段长度多态性(RFLP)检测C1和C2亚型。通过对PCR产物直接测序确定B亚型。对贵州178例B或C基因型HBV感染患者进行检测,其中包括50例无症状携带者(ASC)、100例慢性乙型肝炎(CHB)、14例肝硬化(LC)和14例肝细胞癌(HCC)患者。通过分析患者的HBeAg阳性率、HBV DNA载量和ALT水平,研究HBV C亚型与肝脏疾病进展之间的关系。
84例HBV C基因型患者中,C1亚型27例(32.14%),C2亚型56例(66.67%)。94例B基因型患者中,93例(98.94%)为Ba亚型,仅1例为Bj亚型。C1亚型从ASC和CHB组到LC/HCC组呈逐渐下降趋势。相反,C2亚型在相同顺序中呈逐渐上升(趋势)。C1亚型的HBeAg阳性率显著低于C2亚型。C2亚型患者的ALT水平和HBV DNA载量高于C1亚型患者,但这些指标差异无统计学意义(t = 0.95,0.79)。
贵州以Ba亚型为主,C2亚型较为常见。C1和C2亚型在肝脏疾病各阶段的分布不同。PCR-RFLP方法简便、准确,可用于大规模调查。