慢性乙肝患者HBV核心基因一组单核苷酸多态性与血清HBVDNA水平的相关性研究  

Study on the correlation between a set of single nucleotide polymorphisms of hepatitis B virus core gene and serum HBV DNA levels in patients with chronic hepatitis B

作　　者：李一荣 王雪平 陈凤花 袁琳[1] 马荣红[1] 温晓波[1] 周志明[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院检验科,武汉430022 [2]临床免疫教研室

出　　处：《中华微生物学和免疫学杂志》2011年第5期443-447,共5页Chinese Journal of Microbiology and Immunology

摘　　要：目的研究慢性乙肝患者HBV核心基因的单核苷酸多态性（SNP）与血清HBVDNA水平的相关关系。方法采用PCR-RFLP和限制性内切酶Tsp509I榆测HBV核心基闪的SNP，采用双脱氧终止法对核心基因进行序列测定，实时荧光PCR技术用于HBVDNA水平的定量检测。结果慢性乙肝人群中发现5种典型的PCR-RFLP图谱，分别是RFLP－C、RFLP－D、RFLP－E、RFLP—G和RFLP—C／G，各种RFLPL图谱的分布频率依次为61．5％、2．6％、9．6％、16．7％和9．6％。A165T、A336C、A336T、T337C和T385C等5种SNP与RFLP图谱的变化有关，但是只有SNPA336C和A336T会导致HBcAg第83位的Glu被Asp所替代。RFLP-C组患者的血清HBV DNA水平显著高于RFLP—G组（P=0．02）和RFLP—C／G组（P=0．006），而且RFLP—C／G组患者血清ALT的阳性率显著低于RFLP—C、RFLP—E和RFLP—G组；当HBeAg阳性时，RFLP—C组患者的血清HBVDNA水平显著高于RFLP－G组（P=0．015）和RFLP—C／G组（P=0．008）。结论本研究中使用的PCR—RFLP能够用于检测核心基因的SNP，RFLP—C组患者的血清HBVDNA水平显著高于RFLP—G组和RFLP—C／G组，叮能与Glu83Asp突变有关。Objective To investigate the relation between a set of single nucleotide polymorphisms （SNP） in core gene of HBV in chronic hepatitis B patients and HBV DNA levels. Methods PCR restriction fragment length polymorphism（PCR-RFLP） assay and restriction enzyme Tsp509I were adopted to determine HBV SNP in HBV core gene. Nucleotide sequences of core gene were determined using the dideoxy chain termination method. HBV DNA levels were quantitated with real-time PCR. Results Five typical RFLP patterns, RFLP-C, RFLP-D, RFLP-E, RFLP-G and RFLP-C/G mixture were found and the distribution of HBV RFLP patterns was as follows： C, 61.5% ; D, 2.6% ; E, 9.6% ; G, 16.7% ; C/G mixture, 9.6%. Five SNPs, A165T, A336C, A336T, T337C and T385C, were found to be associated with RFLP patterns change and only SNP A336C or A336T caused the substitution of Glu-83 with Asp in HBcAg. The serum HBV DNA levels in RFLP-C group were higher than that in RFLP-G （P = 0.02 ） and RFLP-C/G group （ P = 0.006 ）, respectively, furthermore, the positive rate of serum ALT in RFLP-C/G group was lower than that in RFLP-C, RFLP-E and RFLP-G group, respectively. Under the condition of HBeAg-positive, the serum HBV DNA levels in RFLP-C group were higher than that in RFLP-G （ P = 0. 015 ） and RFLP-C/G group（ P = 0. 008）, respectively. Conclusion PCR-RFLP used in this study can be adopted to determine HBV SNPs, not genotypes in Chinese patients with chronic hepatitis B. The serum HBV DNA level in RFLP-C group higher than that in RFLP-G or RFLP-C/G group maybe associated with amino acid mutation, Glu83Asp.

关 键 词：乙型肝炎病毒 基因型 聚合酶链反应 多态性 

分 类 号：R512.62[医药卫生—内科学]