赣州地区乙肝病毒基因分型及其临床相关性研究
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摘要
本研究通过收集赣南医学院第一附属医院及赣州市第二人民医院门诊或住院乙肝病毒基因(HBV DNA)检测阳性的慢性乙肝病例303例,采用聚合酶链反应(PCR)-反向点杂交法进行常见基因型别(A-D型)分型检测,同时检测这些病例的乙肝病毒(HBV)血清标志物及肝功能指标,以确定赣州地区HBV基因型的种类及分布频率。通过对HBV不同基因型的乙肝病毒e抗原(HBeAg)阳性率、HBV DNA载量、以及肝功能部分生化指标的比较分析,评估HBV不同基因型别的临床特点。为本地区诊治和预防乙肝提供理论依据。主要研究结果如下：
(1)303例慢性HBV感染者的HBV基因分型结果：B型249例(82.2%),C型39例(12.9%),B／C混合型10例(3.3%),D型3例(1.0%),未分型2例(0.6%),以B、C基因型为主,但B基因型显著多于C基因型(P<0.0001)。说明赣州地区HBV感染存在B基因型、C基因型、B／C混合型及D基因型,此外还有两例未分型病例；赣州地区HBV感染主要以B、C基因型为主,特别以B基因型为优势。
(2)HBV基因型与HBeAg的关系：303例各类乙肝患者的HBV DNA均为阳性,HBeAg总阳性率为46.5%,HBeAg阳性或阴性在B基因型与C基因型中的分布差异无统计学意义(P>0.05),但C基因型HBeAg阳性率要高于B基因型。
(3)HBV基因型与HBV DNA载量的关系：以HBV DNA载量≥Ig5为高载量,B基因型有103例为高载量(103／249,41.4%),而C基因型有34例为高载量(34/39,87.2%),C基因型组HBV DNA高载量病例显著高于B基因型组,差异有统计学意义(P<0.01)。
(4)B基因型、C基因型与肝损害程度之间的关系：对B、C基因型进行ALT、AST、TBA、TBIL、ALB几种生化指标均数间的两两比较,除ALB外(P>0.05),C基因型的其余四种生化指标均显著高于B基因型组(P<0.01或P<0.05)。
通过(2)、(3)、(4)对比分析说明C基因型HBV复制活跃,有更强的致病力,感染者更易发展为严重肝病。
This research is based on the collection of 303 Chronic hepatitis B cases whose HBV DNA tested positive of the outpatients and inpatients in the First Affiliated Hospital of GanNan Medical College and GanZhou City's Second People's Hospital. This study uses the Polymerase ChainReaction(PCR)-Reverse Dot Plot for common genotype(A-D type) and detects HBV serum maker,liver function index to confirm the types and distribution frequency of HBVgenotype in GanZhou area. Through the comparative analysis of the HbeAg's positive frequency, HBV DNA's Bacterial load and the comparison of the biochemical index of the liver function part of the different HBV's genotype, we evaluate clinical features of different HBV genotype, which will provide the theoretical basis to treat and prevent HBV in the local area. The main research results list as follows:
(1) The HBV genotype results of 303 cases Chronic hepatitis B are:B type 249 patients (82.2%), C type 39 patients (12.9%), B/C mixed type 10 patients (3.3%), D type 3 patienst (1.0%), unclassified 2 patients (0.6%). The main type is B, C type, but obviously the B type is more than the C type (P<0.0001). It also shows that there is B genotype,C genotype, B/C mixed genotype, D genotype, and another two unclassified type in HBV infection in GanZhou area. The main HBV infection is B and C type, particularly, the B genotype is in a dominant position.
(2) The relationship between the HBV genotype and HbeAg:the 303 all kinds of hepatitis B patients's HBV DNA tested positive, the total HbeAg positive rate is up to 46.5%, the distribution difference of the HbeAg positive or negative in the B genotype and C genotype has no statistic meaning (P>0.05), but the HbeAg positive rate in C genotype is higher than that of the B genotype.
(3) The relationship between the HBV genotype and the HBV DNA's Bacterial load:the load which is more than Ig5 belongs to high load, and there are 103 cases of B genotype belong to the high load (103/249,41.4%).while there are 34 cases of C genotype (34/39,87.2%). The HBV DNA's Bacterial load of C genotype is obviously higher than B genotype's, and the difference has statistic meaning.
(4) The relationship between B genotype,C genotype and the damage degree of the liver:through the comparisons of the average of biochemical index, the four biochemical indexes of C genotype except ALB are higher than these of B genotype. (P<0.01或P<0.05).
Through the contrast and analysis in article (2)、(3)、(4), we can conclude that the C genotype's HBV has an active replication, and has a strong pathogenicity, so the infected persons are more likely to be a serious disease of liver.
(1)303例慢性HBV感染者的HBV基因分型结果：B型249例(82.2%),C型39例(12.9%),B／C混合型10例(3.3%),D型3例(1.0%),未分型2例(0.6%),以B、C基因型为主,但B基因型显著多于C基因型(P<0.0001)。说明赣州地区HBV感染存在B基因型、C基因型、B／C混合型及D基因型,此外还有两例未分型病例；赣州地区HBV感染主要以B、C基因型为主,特别以B基因型为优势。
(2)HBV基因型与HBeAg的关系：303例各类乙肝患者的HBV DNA均为阳性,HBeAg总阳性率为46.5%,HBeAg阳性或阴性在B基因型与C基因型中的分布差异无统计学意义(P>0.05),但C基因型HBeAg阳性率要高于B基因型。
(3)HBV基因型与HBV DNA载量的关系：以HBV DNA载量≥Ig5为高载量,B基因型有103例为高载量(103／249,41.4%),而C基因型有34例为高载量(34/39,87.2%),C基因型组HBV DNA高载量病例显著高于B基因型组,差异有统计学意义(P<0.01)。
(4)B基因型、C基因型与肝损害程度之间的关系：对B、C基因型进行ALT、AST、TBA、TBIL、ALB几种生化指标均数间的两两比较,除ALB外(P>0.05),C基因型的其余四种生化指标均显著高于B基因型组(P<0.01或P<0.05)。
通过(2)、(3)、(4)对比分析说明C基因型HBV复制活跃,有更强的致病力,感染者更易发展为严重肝病。
This research is based on the collection of 303 Chronic hepatitis B cases whose HBV DNA tested positive of the outpatients and inpatients in the First Affiliated Hospital of GanNan Medical College and GanZhou City's Second People's Hospital. This study uses the Polymerase ChainReaction(PCR)-Reverse Dot Plot for common genotype(A-D type) and detects HBV serum maker,liver function index to confirm the types and distribution frequency of HBVgenotype in GanZhou area. Through the comparative analysis of the HbeAg's positive frequency, HBV DNA's Bacterial load and the comparison of the biochemical index of the liver function part of the different HBV's genotype, we evaluate clinical features of different HBV genotype, which will provide the theoretical basis to treat and prevent HBV in the local area. The main research results list as follows:
(1) The HBV genotype results of 303 cases Chronic hepatitis B are:B type 249 patients (82.2%), C type 39 patients (12.9%), B/C mixed type 10 patients (3.3%), D type 3 patienst (1.0%), unclassified 2 patients (0.6%). The main type is B, C type, but obviously the B type is more than the C type (P<0.0001). It also shows that there is B genotype,C genotype, B/C mixed genotype, D genotype, and another two unclassified type in HBV infection in GanZhou area. The main HBV infection is B and C type, particularly, the B genotype is in a dominant position.
(2) The relationship between the HBV genotype and HbeAg:the 303 all kinds of hepatitis B patients's HBV DNA tested positive, the total HbeAg positive rate is up to 46.5%, the distribution difference of the HbeAg positive or negative in the B genotype and C genotype has no statistic meaning (P>0.05), but the HbeAg positive rate in C genotype is higher than that of the B genotype.
(3) The relationship between the HBV genotype and the HBV DNA's Bacterial load:the load which is more than Ig5 belongs to high load, and there are 103 cases of B genotype belong to the high load (103/249,41.4%).while there are 34 cases of C genotype (34/39,87.2%). The HBV DNA's Bacterial load of C genotype is obviously higher than B genotype's, and the difference has statistic meaning.
(4) The relationship between B genotype,C genotype and the damage degree of the liver:through the comparisons of the average of biochemical index, the four biochemical indexes of C genotype except ALB are higher than these of B genotype. (P<0.01或P<0.05).
Through the contrast and analysis in article (2)、(3)、(4), we can conclude that the C genotype's HBV has an active replication, and has a strong pathogenicity, so the infected persons are more likely to be a serious disease of liver.
引文
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[2]Norder H,HammasB,Lofdahl S,et al.Comparisonof the amino acid sequencesof nine different serotypes of hepatitis B surface antigen and genomic classificationof the corresponding hepatitisBvirus strains[J].J Gen Virol,1992,73 (5):1201-1208.
[3]Weber B. Genetic variability of the S gene of hepatitis Bvirus:clinical and diagnostic impact [J]. J Clin Virol,2005,32 (2):102.
[4]Arauzruiz P, Narder H, Robertson H, et al. GenotypeH:a new Amerindian genotype of the hepatitis B virus revealed in central America[J].J Genvirol,2002,83(4):2059.
[5]范金水等.我国八城市HBsAg阳性和阴性乙肝患者的病毒血清型和基因型分析.中国微生物和免疫学杂志[J],1998,18(3)：88-92.
[6]夏国良等.乙型肝炎病毒基因型和血.清亚型在我国部分地区的分布及其特点。中华流行病学杂志,2001,22(5)：348.
[7]Yeh CT, Liaw YF, Ou JH. The arginine-rich domain of hepatitis Bvirus precore and core proteins contains a signal for nuclear transport. J Virol 1990,64(4):6141-6147.
[8]Eckhardt SG, Milich DR, McLachlan A. Hepatitis B virus core antigenhas two nuclear localization sequences in the arginine-rich carboxyl terminus. J Virol 1991,65(6):575-582.
[9]Abbott WG, Geursen A, Fraser JD, et al.The influence of a maternal chronic hepatitis B virus infection on the repertoire of transcribedT-cell receptor beta chain variable region genes in human cordblood. Hepatology 1995,22(3):1034-1039.
[10]Huy TT, Ushijima H, Quang VX, et al. Genotype C of hepatitis B virus can be classified into at least two subgroups[J].J Gen Virol,2004,85 (2):283.
[11]Tanaka Y, Hasegawa I, Kato T, et al. A casecontrol study for differences among hepatitis B virus infections of genotypes A (subtypes Aa and Ae) and D[J]. Hepatology,2008,40(5):747.
[12]Sugauchi F, Kumada H, Sakugawa H, et al. Two subtypes of genotype B(Ba and Bj) of hepatitis B virus inJapan[J].Clin Infect Dis,2006,38(5):1222.
[13]Erhardt A, Blondin D, Hauck K, et al. Response to interferon alfa is hepatitis B virus genotype dependent; genotype A is more sensitive to inter feron than genotype D[J]. Gut,2006, 54(7):1009-1013.
[14]Yuen MF, Sablon E, Tanaka Y, et al. Epidemiological study of hepatitis B virus genotypes, core promoter and precore mutations of chronic hepatitis B infection in Hong Kong[J]. Hepatol,2004,41(2):119-125.
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