發(fā)布時(shí)間：2018-05-30 22:27

  本文選題：HBV + 屠宰豬��； 參考：《大理學(xué)院》2010年碩士論文

【摘要】：目的本研究采用血清學(xué)和組織病理學(xué)技術(shù)對(duì)220份大理市屠宰豬乙型肝炎病毒(swine hepatitis B virus, SHBV)感染情況進(jìn)行檢測(cè),了解SHBV的感染情況、感染病毒的基因型,檢測(cè)(?)SHBV感染后肝臟組織的免疫病理學(xué)變化,以及SHBV感染后在血清學(xué)和組織病理方面與人HBV的關(guān)系。 方法本研究采集大理市屠宰場(chǎng)屠宰豬血液和肝臟標(biāo)本共220份,血液標(biāo)本離心后分離出血清,采用酶聯(lián)免疫吸附實(shí)驗(yàn)(enzyme-linked immunosorbent assay, ELISA)法檢測(cè)SHBV血清標(biāo)志物(HBV marker, HBVM,俗稱“兩對(duì)半”),型特異性PCR檢測(cè)屠宰豬HBV感染的基因型。肝臟標(biāo)本經(jīng)常規(guī)脫水固定后切片各兩份(4μm×2),一份HE常規(guī)染色檢測(cè)病變組織常規(guī)病理變化,一份采用PV-9000免疫組化試劑盒檢測(cè)屠宰豬肝組織HBsAg/HBcAg感染情況。 結(jié)果(1)220份屠宰豬血清標(biāo)本HBVM檢測(cè)結(jié)果：屠宰豬HBV檢出率為59.1%(130/220)；大三陽(yáng)(HBsAg、HBeAg、HBcAb陽(yáng)性)陽(yáng)性率0.9%(2/220)；其中HBsAg陽(yáng)性23份,陽(yáng)性率為10.5%(23/220)；全陰性率為40.9%(90/220)。(2)型特異性PCR檢測(cè)結(jié)果：選取HBVM陽(yáng)性及免疫組化陽(yáng)性反應(yīng)標(biāo)本(包括全部HBsAg陽(yáng)性標(biāo)本)67份進(jìn)行PCR擴(kuò)增反應(yīng)。凝膠電泳后36份標(biāo)本檢測(cè)出特異性基因條帶,其中顯示在B基因型區(qū)34例,B+C基因區(qū)2例。其中肝臟HBsAg/HBcAg雙陽(yáng)性豬檢出25份,HBsAg或HBcAg單陽(yáng)性檢出11份,二者差異有統(tǒng)計(jì)學(xué)意義(P0.01)。(3)肝組織病理檢測(cè)結(jié)果：HE染色檢測(cè)肝臟組織病變表現(xiàn)為炎細(xì)胞浸潤(rùn)、空泡變性、肝細(xì)胞變性、壞死等：免疫組化在137份標(biāo)本發(fā)現(xiàn)陽(yáng)性反應(yīng),HBsAg主要為漿膜型,HBcAg主要為核型和核漿混合型。表現(xiàn)與人HBV臨床病理各期類似。 結(jié)論(1)豬HBV感染在大理市屠宰豬中有較高的檢出率,感染基因型以B型為主,與大理地區(qū)人HBV感染主要基因型一致。(2)HBV相關(guān)HBsAg/HBcAg在屠宰豬肝組織中有廣泛存在,且病理表現(xiàn)與各期臨床肝炎相似,使豬做為人HBV研究的動(dòng)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.
【學(xué)位授予單位】：大理學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R512.62;R181.3

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