大理市屠宰猪乙型肝炎病毒感染的分子流行病学研究

发布时间：2018-05-30 22:27

本文选题：HBV + 屠宰猪　；参考：《大理学院》2010年硕士论文

【摘要】：目的本研究采用血清学和组织病理学技术对220份大理市屠宰猪乙型肝炎病毒(swine hepatitis B virus, SHBV)感染情况进行检测,了解SHBV的感染情况、感染病毒的基因型,检测(?)SHBV感染后肝脏组织的免疫病理学变化,以及SHBV感染后在血清学和组织病理方面与人HBV的关系。方法本研究采集大理市屠宰场屠宰猪血液和肝脏标本共220份,血液标本离心后分离出血清,采用酶联免疫吸附实验(enzyme-linked immunosorbent assay, ELISA)法检测SHBV血清标志物(HBV marker, HBVM,俗称“两对半”),型特异性PCR检测屠宰猪HBV感染的基因型。肝脏标本经常规脱水固定后切片各两份(4μm×2),一份HE常规染色检测病变组织常规病理变化,一份采用PV-9000免疫组化试剂盒检测屠宰猪肝组织HBsAg/HBcAg感染情况。结果(1)220份屠宰猪血清标本HBVM检测结果：屠宰猪HBV检出率为59.1%(130/220)；大三阳(HBsAg、HBeAg、HBcAb阳性)阳性率0.9%(2/220)；其中HBsAg阳性23份,阳性率为10.5%(23/220)；全阴性率为40.9%(90/220)。(2)型特异性PCR检测结果：选取HBVM阳性及免疫组化阳性反应标本(包括全部HBsAg阳性标本)67份进行PCR扩增反应。凝胶电泳后36份标本检测出特异性基因条带,其中显示在B基因型区34例,B+C基因区2例。其中肝脏HBsAg/HBcAg双阳性猪检出25份,HBsAg或HBcAg单阳性检出11份,二者差异有统计学意义(P0.01)。(3)肝组织病理检测结果：HE染色检测肝脏组织病变表现为炎细胞浸润、空泡变性、肝细胞变性、坏死等：免疫组化在137份标本发现阳性反应,HBsAg主要为浆膜型,HBcAg主要为核型和核浆混合型。表现与人HBV临床病理各期类似。结论(1)猪HBV感染在大理市屠宰猪中有较高的检出率,感染基因型以B型为主,与大理地区人HBV感染主要基因型一致。(2)HBV相关HBsAg/HBcAg在屠宰猪肝组织中有广泛存在,且病理表现与各期临床肝炎相似,使猪做为人HBV研究的动物模型提供了可能。
[Abstract]:Objective to detect the infection of hepatitis B virus (HBV) hepatitis B virus, SHBV) in 220 pigs slaughtered in Dali by serological and histopathological techniques, and to understand the infection of SHBV and the genotype of the virus. The changes of liver immunopathology and the relationship between the serological and histopathological aspects of SHBV infection and human HBV were detected. Methods A total of 220 blood and liver samples were collected from slaughtered pigs in Dali slaughterhouse. Serum was isolated from blood samples after centrifugation. Enzyme linked immunosorbent assay (Elisa) was used to detect the genotypes of HBV infection in slaughtered pigs. Liver specimens were fixed with routine dehydration and fixed by routine dehydration. The liver sections were divided into 4 渭 m 脳 2 pieces each. One specimen was stained with HE to detect the pathological changes of pathological tissues, and the other one was detected for HBsAg/HBcAg infection in slaughtered pig liver tissue by PV-9000 immunohistochemical kit. Results the results of HBVM detection in 220 serum samples of slaughtered pigs were as follows: the positive rate of HBV in slaughtered pigs was 59.1% (130 / 220); the positive rate of HBcAb in HBeAg of HBeAg was 0.910%; the positive rate of HBcAb was 0.92%; 23 of them were positive for HBsAg. The positive rate was 10.5% 23 / 220% and the total negative rate was 40.9% 90 / 220%) type specific PCR: 67 samples of HBVM positive and immunocytochemical reaction (including all HBsAg positive specimens) were selected for PCR amplification. After gel electrophoresis, specific gene bands were detected in 36 samples, of which 2 were found in B genotype in 34 cases. Of the 25 HBsAg/HBcAg positive pigs, 11 were HBsAg/HBcAg positive or HBcAg positive. The difference between them was significant (P 0.01). The pathological results of liver tissue were as follows: the pathological changes were inflammatory cell infiltration, vacuolar degeneration and hepatocyte degeneration. Necrosis: the positive reaction of HBsAg was serous type HBcAg was mainly nuclear and cytoplasmic mixed type in 137 specimens by immunohistochemistry. The findings were similar to those of human HBV in clinical and pathological stages. Conclusion (1) the positive rate of HBV infection in Dali slaughtered pigs is high. The genotype of HBV infection is mainly type B, which is consistent with the main genotypes of HBV infection in Dali area. The pathological findings are similar to those of clinical hepatitis, which makes it possible for pigs to be used as animal models of human HBV.
【学位授予单位】：大理学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R512.62;R181.3

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