慢乙肝患者HBVcccDNA与血清HBVDNA及其标志物相关性研究

发布时间：2018-08-22 07:23

【摘要】：目的探讨慢性乙型病毒性肝炎（CHB）患者肝组织HBVcccDNA与血清HBV DNA及其标志物等之间的相关性。方法采用实时定量荧光PCR检测法检测10例CHB患者肝组织HBVcccDNA，用PCR-荧光探针法检测血清HBV DNA含量，用电化学发光免疫分析法定量检测乙肝标志物，常规病理学染色分析病理学改变，对肝组织进行炎症活动度分级(G)及纤维化程度分期(S)。结果①10例CHB患者肝组织病理结果：G2S1：3例；G2S2：2例；G2S3：1例；G3S2：2例；G3S3：2例。②10例CHB患者肝组织均检测到HBVcccDNA，定量值为1.80x104拷贝/mL～8.50x109拷贝/mL。10例CHB患者血清HBVDNA:阳性8例，定量值为2.23xlO5拷贝/mL～3.08xlO8拷贝/mL；阴性2例(低于检测下限103拷贝/mL)。③肝组织HBVcccDNA定量值与肝组织炎症活动度(G)及纤维化程度(S)无相关性（Ｐ0.05）。④肝组织HBVcccDNA定量值与血清HBV DNA定量值成高度正相关（ｒ=0.898，Ｐ0.01）。⑤肝组织HBVcccDNA定量值与血清HBeAg滴度呈高度正相关性(r=0.821, Ｐ0.01)，与血清HBsAg滴度无相关性(r=-0.483, Ｐ0.05）。⑥肝组织HBVcccDNA定量值与ALT、AST定量水平均无相关性。⑦血清HBV DNA与HBsAg、HBeAg、ALT及AST在统计学上，均无相关性（0.05）。结论肝组织HBVcccDNA定量值与血清HBV DNA定量值和HBeAg滴度呈高度正相关，均能够直接反映CHB患者体内HBV的存在和复制状态；但其定量值的高低与肝组织病理结果、HBsAg滴度、ALT、AST水平均无相关。肝组织HBVcccDNA定量检测联合血清HBVDNA和HBeAg是判断抗病毒治疗疗效的可靠指标。目的探讨慢性乙型病毒性肝炎（CHB）患者血清HBVcccDNA与血清HBV DNA及其标志物等之间的相关性。方法采用实时定量荧光PCR检测法检测10例CHB患者血清HBVcccDNA，用PCR-荧光探针法检测血清HBV DNA含量，用电化学发光免疫分析法定量检测乙肝标志物。结果①10例CHB患者血清中检测到HBVcccDNA有6例，定量值为4.68x103拷贝/mL～1.08x107拷贝/mL，，未检测到者4例(低于检测下限103拷贝/mL)。②10例CHB患者血清HBV DNA阳性10例，定量值为3.51xlO5拷贝/mL～6.29xlO8拷贝/mL。③CHB血清HBVcccDNA定量值与血清HBV DNA定量值、血清HBeAg滴度、血清HBsAg滴度均没有显著性的相关性（ｒ=0.049，Ｐ0.05；r=0.302,0.05；r=-0.248, Ｐ0.05）。④血清HBVcccDNA定量值与ALT、AST水平均无相关性(Ｐ0.05）。结论血清HBVcccDNA水平与血清HBVDNA水平、血清HBSAg和ＨＢeＡｇ的滴度以及ALT、AST均无相关。由此可见检测血清HBVcccDNA的定量并不能全面的反应血中HBV的复制和感染。所以不能成为血清HBVDNA及乙肝血清标志物的替代指标。目的探讨慢性乙型病毒性肝炎（CHB）在未进行抗病毒之前检测患者P-S区基因变异。方法采用实时定量荧光PCR检测法检测P-S区基因变异。结果10例CHB患者血清均未检测到P-S区基因变异。结论本研究没有检测到慢乙肝患者在接受抗病毒之前发生基因突变导致的原发性耐药，因此仍需要以后的研究实验进一步证实。
[Abstract]:Objective to investigate the relationship between HBVcccDNA in liver tissue and serum HBV DNA and its markers in patients with chronic hepatitis B (CHB). Methods Hepatic HBV cccDNAs were detected by real-time quantitative PCR assay, serum HBV DNA levels were detected by PCR-fluorescence probe method, HBV markers were quantitatively detected by electrochemiluminescence immunoassay, pathological changes were analyzed by routine pathological staining. Grade of inflammation activity (G) and stage of fibrosis (S). For liver tissue Results the histopathological findings of liver tissue of 110 CHB patients were as follows: 1 case of G2S2: 1 case of G3S2: 1 case of G2S2: 1 case of G2S2 HBV cccDNAwas detected in liver tissue of 210 CHB patients with G3S3:2. The quantitative value was 1.80x104 copy / mLX 8.50x109 / mL.10. The serum HBV DNA was positive in 8 cases and 2.23xlO5 copy / mLX 3.08xlO8 copy / mLL in the serum of CHB patients. There was no significant correlation between the quantitative value of liver tissue HBVcccDNA and the degree of hepatic inflammation (G) and fibrosis (S) (P0.05). There was a high positive correlation between the quantitative value of liver tissue HBVcccDNA and serum HBV DNA (P0.05). The quantitative value of HBVcccDNA in liver tissue was positively correlated with the titer of serum HBeAg (r = 0.821, P 0.01), but not correlated with the titer of serum HBsAg (r-0.483, P0.05) .6 there was no correlation between the quantitative value of HBVcccDNA in liver tissue and the quantitative level of alt. 7. There was no statistical correlation between serum HBV DNA and HBHBeAgALT and AST. There was no correlation (0.05). Conclusion there is a highly positive correlation between the quantitative value of HBVcccDNA in liver tissue and the serum HBV DNA and HBeAg titer, which can directly reflect the existence and replication of HBV in patients with CHB, but there is no correlation between the quantitative value of CHB and the level of serum HBV DNA titer and alt level in patients with CHB. The quantitative detection of liver tissue HBVcccDNA combined with serum HBVDNA and HBeAg is a reliable index for evaluating the efficacy of antiviral therapy. Objective to investigate the relationship between serum HBVcccDNA, serum HBV DNA and its markers in patients with chronic hepatitis B (CHB). Methods Serum HBV cccDNAs in 10 patients with CHB were detected by real-time quantitative fluorescence PCR assay, serum HBV DNA levels were detected by PCR-fluorescence probe method, and HBV markers were quantitatively detected by electrochemiluminescence immunoassay (ECLIA). Results there were 6 cases of HBVcccDNA detected in the serum of 110 patients with CHB. The quantitative value was 1.08x107 copies / mLof 4.68x103 copy. 4 cases (below the detection limit of 103copies / mL) and 10 cases of CHB patients were positive for HBV DNA in the serum of 4 cases (below the detection limit of 103copies / mL). There was no significant correlation between the quantitative value of serum HBVcccDNA and serum HBV DNA, the titer of serum HBeAg and the titer of serum HBsAg (r 0.049%, P 0.05 P 0.05 0. 302 卤0. 05). There was no correlation between serum HBVcccDNA and alt level (P0.05). Conclusion there is no correlation between serum HBVcccDNA level and serum HBVDNA level, serum HBSAg and HBeAg titers and alt AST. It can be seen that the detection of serum HBVcccDNA does not fully respond to the replication and infection of HBV in the blood. So it can not be a substitute index of serum HBVDNA and hepatitis B serum marker. Objective to investigate the gene variation of P-S region in patients with chronic viral hepatitis B (CHB) before antiviral detection by (CHB). Methods A real-time quantitative PCR assay was used to detect the gene variation of P-S region. Results No P-S gene mutation was detected in serum of 10 patients with CHB. Conclusion this study did not detect the primary drug resistance caused by gene mutation in patients with chronic hepatitis B before receiving antiviral therapy.
【学位授予单位】：宁夏医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.62

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