CHB患者HBV基因型及耐药突变基因联合检测临床研究

Clinical Study of Combined Detection about Hepatitis B Virus Genotypes and Drug Resistant Mutation Genes in Patients with Chronic Hepatitis B

目的 研究慢性乙型肝炎（chronic hepatitis B,CHB）患者乙型肝炎病毒（Hepatitis B virus,HBV）基因型分布特征,监测HBV发生耐药突变基因状况.方法 随机收集CHB患者血清样本共52例,其中男性32例,女性20例.采用实时荧光定量聚合酶链式反应检测血清HBV DNA含量.通过聚合酶链式反应（polymerase chain reaction,PCR）和反向斑点（reverse dot blot,RDB）杂交技术联合检测HBV基因型以及多位点耐药突变基因,并对受检者相关资料进行分析,统计学分析所用软件为GraphPad Prism 5.0.结果 男性患者平均HBV DNA含量5.38±1.15 lg IU/ml,女性患者平均HBV DNA含量5.05±1.59 lg IU/ml.不同性别组之间HBV DNA含量差异无统计学意义（t=0.875 0,P=0.3857＞0.05）.HBV C基因型检出率最高48.08％ （25/52）,其次为B基因型36.54％（19/52）.总耐药突变率30.77％（16/52）,rt204位点发生突变率最高50.0％（8/16）.HBV不同基因型之间不同位点发生耐药突变频率差异无统计学意义（χ2 =7.286,P=0.6982＞0.05）.结论 C型为优势流行基因型;rt204为最易产生耐药突变基因位点;联合检测HBV基因分型和耐药突变基因具有重要临床意义.

Objective To investigate the distribution of hepatitis B virus（HBV） genotypes and monitor the frequency of drug resistant mutations in patients with chronic hepatitis B. Methods Conducted a cohort study with 52 participants including 32 male patients and 20 female patients with CHB. HBV DNA was detected by real time fluorescent quantitative polymerase chain reaction. HBV genotypes and drug resistant mutations were simultaneously observed by polymerase chain reaction and reverse dot blot （RDB）. And the related data of all subjects were statistically analyzed by GraphPad Prism 5.0 software. Re- sults The average degree of HBV DNA in male patients with CHB was 5.38±1.15 lg IU/ml, and the average degree of HBV DNA in female patients with CHB was 5.05 ± 1.59 lg IU/ml. There was no statistically significance difference be- tween the different gender groups （t=0. 8750 ,P=0. 3857〉0.05）. Detection ratios of genotype C and B were 48.08% （25/ 52） and 36.54%（19/52） respectively. The total ratio of drug resistant mutations was 30.77% （16/52）. Drug resistant mu- tations were frequently detected in gene site rt204 （8/16,50.0%）. There was no statistically significant difference （χ2 = 7. 286,P=0. 6982〉0.05） for drug resistant mutation ratios in different gene sites among the different genotype groups. Conclusion HBV genotype C was the predominant genotype. Drug resistant mutations were easily taken place in gene site rt204. It was very significant to detect HBV genotypes and drug resistance mutations for clinical treatments.