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鳗鲡匹里虫小亚基核糖体RNA基因序列及用于该寄生虫诊断检测的PCR引物

The small subunit ribosomal RNA gene sequence of Pleistophora anguillarum and the use of PCR primers for diagnostic detection of the parasite.

作者信息

Hung H W, Lo C F, Tseng C C, Peng S E, Chou C M, Kou G H

机构信息

Department of Zoology, National Taiwan University, Taipei, ROC.

出版信息

J Eukaryot Microbiol. 1998 Sep-Oct;45(5):556-60. doi: 10.1111/j.1550-7408.1998.tb05116.x.

Abstract

Using the polymerase chain reaction (PCR) and two primers for conserved regions of the small subunit ribosomal RNA (SSU-rRNA) of Microsporidia, a DNA segment about 1,195 base pairs long was amplified from a DNA template prepared from purified spores of the microsporidian species Pleistophora anguillarum. These spores had been isolated from adult eels (Anguilla japonica) with "Beko Disease." A comparison of sequence data from other microsporidian species showed P. anguillarum SSU-rRNA to be most similar to Vavraia oncoperae. When juvenile eels were artificially infected with P. anguillarum, enzyme-linked immunosorbent assay could detect a positive infection only 12 days post-infection. However, when suitable PCR primers were used, a DNA fragment of about 0.8 kb was detected from these juvenile eels after only 3 days post infection. No PCR product was obtained with templates prepared from clinically healthy control animals.

摘要

利用聚合酶链反应(PCR)以及针对微孢子虫小亚基核糖体RNA(SSU-rRNA)保守区域的两种引物,从由微孢子虫物种鳗鲡匹里虫(Pleistophora anguillarum)纯化孢子制备的DNA模板中扩增出一段约1195个碱基对长的DNA片段。这些孢子是从患有“贝科病”的成年鳗鱼(日本鳗鲡,Anguilla japonica)中分离得到的。与其他微孢子虫物种的序列数据比较显示,鳗鲡匹里虫的SSU-rRNA与肿瘤瓦夫拉虫(Vavraia oncoperae)最为相似。当幼鳗被鳗鲡匹里虫人工感染时,酶联免疫吸附测定在感染后仅12天才能检测到阳性感染。然而,当使用合适的PCR引物时,在感染后仅3天就从这些幼鳗中检测到约0.8 kb的DNA片段。从临床健康对照动物制备的模板未获得PCR产物。

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