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两种检测实验性诱导感染猪粪便中传染性胃肠炎病毒方法的比较。

Comparison of two methods for detection of transmissible gastroenteritis virus in feces of pigs with experimentally induced infection.

作者信息

van Nieuwstadt A P, Cornelissen J B, Zetstra T

机构信息

Virology Department, Central Veterinary Institute, Lelystad, The Netherlands.

出版信息

Am J Vet Res. 1988 Nov;49(11):1836-43.

PMID:2854707
Abstract

An indirect, double-antibody sandwich-type ELISA for detection of transmissible gastroenteritis virus (TGEV) was developed, using a solid phase of rabbit hyperimmune serum and a pool of 3 antipeplomer monoclonal antibodies to trap and to detect the virus, respectively. The technique was used to detect viral antigen in feces of pigs that had been infected with the virulent Miller strain, the attenuated Purdue strain, or the Erica strain (a Dutch field isolate) of TGEV. The results were compared with those of a solid-phase immunosorbent electron microscopy (SPIEM) technique for virus detection. Both techniques detected shedding of virulent virus in feces obtained from pigs on the first or second day after infection, and virus excretion continued for 6 to 8 consecutive days. Virus shedding started later in pigs infected with the attenuated Purdue strain of TGEV and lasted only 2 to 4 days. In comparison with the 2 virulent strains, infection with the attenuated strain appeared to be limited to a smaller portion of the small intestine. Of 242 fecal specimens that were tested by use of ELISA and SPIEM, 119 had positive results in both tests. Additionally, virus could be detected by ELISA in 21 and by SPIEM in 16 specimens. Fecal specimens obtained from pigs before infection always reacted negatively by ELISA for TGEV antigen; there was no cross-reactivity with fecal specimens containing porcine rotavirus or porcine epidemic diarrhea virus. The ELISA and SPIEM were found to be specific and sensitive for the detection of TGEV in feces.

摘要

开发了一种间接双抗体夹心型酶联免疫吸附测定法(ELISA)用于检测传染性胃肠炎病毒(TGEV),该方法使用兔超免疫血清作为固相,并用一组3种抗纤突蛋白单克隆抗体分别捕获和检测病毒。该技术用于检测感染了TGEV强毒株米勒株、弱毒普渡株或埃丽卡株(一种荷兰田间分离株)的猪粪便中的病毒抗原。将结果与用于病毒检测的固相免疫吸附电子显微镜(SPIEM)技术的结果进行比较。两种技术均检测到感染后第1天或第2天从猪获得的粪便中有强毒病毒排出,并且病毒排泄持续6至8天。感染TGEV弱毒普渡株的猪病毒排出开始较晚,仅持续2至4天。与2种强毒株相比,感染弱毒株似乎局限于小肠的较小部分。在使用ELISA和SPIEM检测的242份粪便标本中,119份在两种检测中均呈阳性结果。此外,ELISA可在21份标本中检测到病毒,SPIEM可在16份标本中检测到病毒。感染前从猪获得的粪便标本ELISA检测TGEV抗原总是呈阴性反应;与含有猪轮状病毒或猪流行性腹泻病毒的粪便标本无交叉反应。发现ELISA和SPIEM对检测粪便中的TGEV具有特异性和敏感性。

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