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Detetion of Hepatitis E Virus in Pig Products from Abattoirs and Establishment of HEV Infection Model for Rats

Author: GongGa
Tutor: CuiLi;HuaXiuGuo
School: Shanghai Jiaotong University
Course: Preventive Veterinary Medicine
Keywords: Swine hepatitis E virus RT-PCT detection method Pig slaughtering products Rat infection model
CLC: S855.3
Type: Master's thesis
Year: 2009
Downloads: 55
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Hepatitis E virus (HEV) is a fecal - oral transmission Zoonosis pathogens. Numerous studies indicate that the pig is the natural host for HEV Although HEV infection is not caused by pigs obvious clinical symptoms, but HEV It is through this natural host a large number of copy storage, which infects humans, and poses a potential threat to human public health. This study proposes the laboratory tests of a swine HEV HEV carrying pig slaughtering products, the use of the method and the the SD rats HEV infection model. The research for animal inspection and quarantine departments The swine HEV monitoring, diagnosis, such as technical support and basic theoretical basis. , Of swine HEV laboratory testing method to establish the hepatitis E is mainly detected by ELASA (enzyme-linked immunosorbent) Act HEV antibody diagnostic window period, but HEV infection, the patient's own immune system and ELASA reagents itself many reasons to amyl the diagnosis of liver difficulty. HEV RNA detection is to determine the the HEV virus exists standard one. In this study, using two PCR detection methods to detect swine HEV RNA. Degenerate primers specific fragment has been submitted to the GenBank pig HEVORF2 design nested successfully established a laboratory swine HEV RT-nPCR detection method. The results show that this method has a high degree of specificity and sensitivity, and can be used in pig slaughter scene investigation. Submitted in GenBank pig HEVORF3 specific fragment primers were designed to establish the swine HEV Tag Man probe fluorescence quantitative PCR method. The method is simple, rapid, specific advantages. Two, the pig slaughter Product HEV carry 120 pig bile collected from the Shanghai area slaughterhouse survey of this study, 120 pig liver, 120 pig spleen, 120 pig mesenteric lymph nodes, a total of 729 of the 126 pig feces, 123 pig blood samples using RT-nPCR method established, HEV RNA samples testing positive amplicon TA cloning sequencing and phylogenetic analysis of viral sequences obtained results indicate that the slaughterhouse pigs carry the HEV present in the bile, serum and stool samples. Phylogenetic analysis showed that the slaughterhouse pig products HEV both genotype 3 have genotype 4 HEV strains. The results suggest that the slaughterhouse must properly handle the time of slaughter, serum, bile and feces, prevent HEV pollution from other slaughter products. Third, of swine HEV rat infection model to establish this study, HEV-positive pig manure supernatant inoculated SPF SD rats via the tail vein, HEV RNA was detected by RT-nPCR using indirect immunofluorescence observed HEV pathogen in the rat the dynamic distribution of the organization, while taking advantage of the pathological changes in histopathology observation challenged rats of different organizations. Rats infected with HEV obvious symptoms, can be observed in the liver, spleen, lymph node HEV antigen can be detected in the serum, stool HEV RNA, liver, spleen, lymph nodes, pathological changes, and in the late stage of infection can be detected HEV antibodies, but there is no consistency to enhance the activity of serum ALT. TBIL level during the whole experiment has been at normal levels, indicating that HEV infection does not affect the level of bilirubin. This study proposes two swine HEV PCR detection method - RT-nPCR assay and Tag Man probe fluorescence quantitative PCR method, and the establishment of the SPF rats infected with HEV model rats infected with HEV histopathological changes HEV in different tissue distribution, and virus RNA, serum transaminases and serum antibodies at different times of change. Preliminary theoretical and technical reserves to prevent the outbreak of hepatitis E disease, measures to develop comprehensive prevention and control of hepatitis E disease is of great significance.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Veterinary Infectious Diseases > Virus disease
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