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改良聚合酶链反应方案与间接血凝试验在检测猪附红细胞体感染中的比较

Comparison of an improved polymerase chain reaction protocol and the indirect hemagglutination assay in the detection of Eperythrozoon suis infection.

作者信息

Gwaltney S M, Oberst R D

机构信息

Department of Veterinary Diagnostic Investigation, College of Veterinary Medicine, Kansas State University, Manhattan 66506.

出版信息

J Vet Diagn Invest. 1994 Jul;6(3):321-5. doi: 10.1177/104063879400600307.

Abstract

The ability of an improved polymerase chain reaction (PCR) protocol to detect Eperythrozoon suis DNA in the blood of experimentally infected nonsplenectomized pigs was evaluated. The protocol utilizes previously described E. suis-specific primers and a proprietary DNA-releasing reagent in a 2-step amplification cycle followed by visualization of the 492-bp amplification product on agarose gels. This PCR protocol successfully amplified E. suis DNA in blood from all postinfection samples and from the preinfection samples of 2 pigs, indicating preexisting natural infections. Results of the indirect hemagglutination test on serum samples from these pigs revealed that only 1 pig developed detectable antibody titers to E. suis infection during the 43-day study; that pig was determined by PCR to have been infected naturally with E. suis prior to experimental inoculation. These results confirm previous reports of poor antibody response of young pigs to E. suis infection and demonstrate the potential of PCR as a valuable tool for the diagnosis and study of E. suis infection in pigs.

摘要

对一种改良的聚合酶链反应(PCR)方法在检测实验感染但未进行脾切除的猪血液中猪附红细胞体DNA的能力进行了评估。该方法在两步扩增循环中使用先前描述的猪附红细胞体特异性引物和一种专利DNA释放试剂,随后在琼脂糖凝胶上对492 bp的扩增产物进行可视化分析。此PCR方法成功扩增了所有感染后样本以及2头猪感染前样本血液中的猪附红细胞体DNA,表明存在自然感染。对这些猪血清样本进行的间接血凝试验结果显示,在为期43天的研究中,只有1头猪产生了可检测到的针对猪附红细胞体感染的抗体效价;通过PCR确定该猪在实验接种前已自然感染猪附红细胞体。这些结果证实了先前关于幼猪对猪附红细胞体感染抗体反应不佳的报道,并证明了PCR作为诊断和研究猪附红细胞体感染的有价值工具的潜力。

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