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通过聚合酶链反应(PCR)检测自然发生的牛流产胎儿组织中的犬新孢子虫。

Detection by PCR of Neospora caninum in fetal tissues from spontaneous bovine abortions.

作者信息

Baszler T V, Gay L J, Long M T, Mathison B A

机构信息

Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164-7040, USA.

出版信息

J Clin Microbiol. 1999 Dec;37(12):4059-64. doi: 10.1128/JCM.37.12.4059-4064.1999.

Abstract

The routine diagnosis of Neospora caninum abortion is based upon histopathologic changes in fetal tissues and identification of tissue parasites by immunohistochemistry. Confirmation of N. caninum infection by immunohistochemistry has low sensitivity. In the present study, we examined the utility of PCR in detecting N. caninum infection in fetal tissues from spontaneous bovine abortion. DNA was obtained from fresh and formalin-fixed tissues from 61 bovine fetuses submitted for abortion diagnosis. Histopathology and immunohistochemistry determined the true status of N. caninum infection in each fetus. In formalin-fixed paraffin-embedded tissues, PCR detected N. caninum DNA in 13 of 13 true-positive fetuses (100%) and in 1 of 16 true-negative fetuses (6%). In fresh or frozen tissues, PCR detected N. caninum DNA in 10 of 13 true-positive fetuses (77%) and 0 of 11 true-negative fetuses (0%). PCR also detected N. caninum DNA in 6 of 8 fetuses that had typical lesions of N. caninum but were immunohistochemistry negative, indicating a higher sensitivity of PCR in comparison to that of immunohistochemistry. N. caninum DNA was amplified most consistently from brain tissue. PCR detection of N. caninum DNA in formalin-fixed, paraffin-embedded tissues was superior to that in fresh tissues, presumably because of the increased accuracy of sample selection inherent in histologic specimens.

摘要

犬新孢子虫流产的常规诊断基于胎儿组织的组织病理学变化以及通过免疫组织化学鉴定组织中的寄生虫。通过免疫组织化学确认犬新孢子虫感染的敏感性较低。在本研究中,我们检测了PCR在检测自然流产牛胎儿组织中犬新孢子虫感染方面的效用。从提交用于流产诊断的61个牛胎儿的新鲜和福尔马林固定组织中获取DNA。组织病理学和免疫组织化学确定了每个胎儿中犬新孢子虫感染的真实状况。在福尔马林固定石蜡包埋组织中,PCR在13个真阳性胎儿中的13个(100%)以及16个真阴性胎儿中的1个(6%)中检测到犬新孢子虫DNA。在新鲜或冷冻组织中,PCR在13个真阳性胎儿中的10个(77%)以及11个真阴性胎儿中的0个(0%)中检测到犬新孢子虫DNA。PCR还在8个具有典型犬新孢子虫病变但免疫组织化学呈阴性的胎儿中的6个中检测到犬新孢子虫DNA,这表明与免疫组织化学相比,PCR具有更高的敏感性。从脑组织中最一致地扩增出犬新孢子虫DNA。在福尔马林固定石蜡包埋组织中PCR检测犬新孢子虫DNA优于新鲜组织,这可能是因为组织学标本中样本选择的准确性提高。

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