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聚合酶链反应(PCR)与其他检测方法在犬埃立克体病早期诊断中的比较。

Comparison of PCR with other tests for early diagnosis of canine ehrlichiosis.

作者信息

Iqbal Z, Chaichanasiriwithaya W, Rikihisa Y

机构信息

Department of Veterinary Pathobiology, College of Veterinary Medicine, Ohio State University, Columbus 43210.

出版信息

J Clin Microbiol. 1994 Jul;32(7):1658-62. doi: 10.1128/jcm.32.7.1658-1662.1994.

Abstract

The purpose of the study was to compare the sensitivity of PCR with those of cell culture reisolation of Ehrlichia canis, the indirect fluorescent antibody test (IFA), and Western immunoblotting (WI) in the early diagnosis of canine ehrlichiosis. Five German shepherd dogs were intravenously inoculated with 10(7) E. canis-infected DH82 cells. Blood was collected on alternate days during a 2-week postinoculation period. Mononuclear cell fractions were harvested and used for E. canis reisolation and DNA extraction for PCR. The plasma was used for assaying antibodies against E. canis. By PCR, the 16S rRNA gene of E. canis was detected in the mononuclear cell specimens collected as early as day 4 to 10 postexposure (PE). E. canis was reisolated from the blood starting on day 2 PE from all five dogs. The indirect fluorescent antibody test and Western immunoblotting could detect E. canis antibodies as early as 2 to 8 days PE. Cell culture reisolation proved to be the most sensitive and definitive for early diagnosis of ehrlichiosis, but it is not very convenient, since it takes a long time (14 to 34 days) to show up positive. The sensitivity of PCR is comparable to or slightly less than that of other established methods; however, the convenience, quickness, and direct nature of detecting E. canis DNA is expected to make PCR more useful for clinical diagnosis.

摘要

本研究的目的是比较聚合酶链反应(PCR)与犬埃立克体细胞培养再分离、间接荧光抗体试验(IFA)及蛋白质免疫印迹法(WI)在犬埃立克体病早期诊断中的敏感性。五只德国牧羊犬静脉接种10⁷个感染犬埃立克体的DH82细胞。在接种后2周内每隔一天采集血液。收集单核细胞组分用于犬埃立克体的再分离及PCR的DNA提取。血浆用于检测抗犬埃立克体抗体。通过PCR,在暴露后(PE)第4至10天采集的单核细胞标本中检测到犬埃立克体的16S rRNA基因。从所有五只犬的PE第2天开始,血液中再次分离出犬埃立克体。间接荧光抗体试验和蛋白质免疫印迹法最早可在PE第2至8天检测到犬埃立克体抗体。细胞培养再分离被证明是埃立克体病早期诊断中最敏感和最具决定性的方法,但不太方便,因为出现阳性结果需要很长时间(14至34天)。PCR的敏感性与其他既定方法相当或略低;然而,检测犬埃立克体DNA的便利性、快速性和直接性有望使PCR在临床诊断中更有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/03f1/263754/87c7a587557a/jcm00007-0051-a.jpg

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