摘要
目的通过用人工繁育的围生期和幼年树鼩接种人乙型肝炎病毒(HBV),并优选感染指标的检测方法,提高树鼩对HBV的感染率及检出方法的敏感性和可靠性。方法用人HBV阳性血清接种42只人工繁育的围生期和幼年树鼩,选择合适的引物,经巢式PCR(nPCR)技术检测它们肝组织及血清的HBV DNA,并对扩增出的DNA片段进行测序验证,以及对活检肝组织作病理组织学观察。结果接种后动物的肝组织和血清HBV DNA的阳性率分别为47.6%(20/42)和71.4%(30/42),肝组织和血清共同阳性的为40.5%(17/42),总阳性率为78.6%(33/42);扩增的DNA片段经测序与人HBV DNA相应片段的同源性为98%和99%;肝组织病理改变以轻度炎性病变较多见。结论用人工繁育的围生期和幼年树鼩感染HBV,可提高树鼩对HBV的感染效率;用nPCR技术及选择合适的引物配对检测肝组织和血清标本的HBV DNA,是评价树鼩感染HBV状况的敏感、可靠方法。
To improve the efficiency of using the artificially fed and young tree shrews as the infection model for human hepatitis B virus （HBV） as well as the sensitivity and reliability of this method as the detection marker in the samples of infected animals, 42 artificially fed perinatal and young tree shrews were inoculated with positive human HBV serum, and nested polymerase chain reaction （nPCR） with optimized primers was used to detect HBV DNA in samples of liver biopsy and serum specimens. The amplified products from nPCR were sequenced and compared with the corresponding sequences of HBV genome.s, and the liver biopsies were observed histopathologically. It was demonstrated that the positive rates of HBV DNA detected in liver biopsy samples and serum from the infected pups as revealed by nPCR with proper primers were 47.6 % （20/42） and 71.4% （30/42） respectively, with the total positive rate of 78.6% （33/42）, in which 40.5% （17/42） of pups showed the presence of HBV DNA both in liver biopsy and serum samples. The sequences amplified from samples of the tree shrews were similar to those of the corresponding fragments of HBV genomes, with the homology of 98% and 99 %. The main pathological change in the liver biopsies was mild inflammation. It is concluded that application of the artificially fed perinatal and young tree shrews as the infection model can improve the infection efficiency of HBV to tree shrews, and nPCR technique with proper primers is fairly useful for precisely detecting HBV DNA in the infected tree shrew.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2006年第8期792-795,共4页
Chinese Journal of Zoonoses
基金
广西科学基(金桂科回0144002
桂科自0447087和桂科回0236008)资助
