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使用聚合酶链反应检测猪附红细胞体。

Detection of Eperythrozoon suis using the polymerase chain reaction.

作者信息

Gwaltney S M, Hays M P, Oberst R D

机构信息

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

出版信息

J Vet Diagn Invest. 1993 Jan;5(1):40-6. doi: 10.1177/104063879300500110.

Abstract

Eperythrozoon suis is an extracellular red blood cell parasite that causes icteroanemia in acutely ill pigs and a variety of syndromes in chronically infected pigs. Current techniques to detect E. suis infection are limited by variability of parasitemias and antibody responses in infected animals. The polymerase chain reaction (PCR) was investigated to determine its potential as a means of detecting E. suis infection in pigs. With DNA samples extracted from either purified E. suis organisms or E. suis-infected pig blood, PCR produced an amplification product 492 base pairs in length. This amplification product hybridized successfully with the fragment of the DNA probe from which the primer sequences had been selected. Sensitivity studies indicated that the PCR protocol was capable of amplifying total genomic E. suis DNA in quantities as low as 450 pg. When PCR was used with DNA from blood samples from a splenectomized pig that had been infected with E. suis, amplification products were detectable as early as 24 hours postinfection. This preliminary analysis indicates that PCR shows promise as a means of efficiently detecting E. suis infection in pigs.

摘要

猪附红细胞体是一种细胞外红细胞寄生虫,可导致急性发病猪出现黄疸性贫血,并在慢性感染猪中引发多种综合征。目前检测猪附红细胞体感染的技术受到感染动物中虫血症和抗体反应变异性的限制。对聚合酶链反应(PCR)进行了研究,以确定其作为检测猪附红细胞体感染方法的潜力。用从纯化的猪附红细胞体生物体或感染猪附红细胞体的猪血中提取的DNA样本进行PCR,产生了一个长度为492个碱基对的扩增产物。该扩增产物与从中选择引物序列的DNA探针片段成功杂交。敏感性研究表明,该PCR方案能够扩增低至450 pg的猪附红细胞体总基因组DNA。当将PCR用于来自感染猪附红细胞体的脾切除猪的血液样本的DNA时,早在感染后24小时就可检测到扩增产物。这一初步分析表明,PCR有望成为一种有效检测猪附红细胞体感染的方法。

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