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首页> 美国卫生研究院文献>Journal of Clinical Microbiology >A cloned DNA probe for Cowdria ruminantium hybridizes with eight heartwater strains and detects infected sheep.
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A cloned DNA probe for Cowdria ruminantium hybridizes with eight heartwater strains and detects infected sheep.

机译：一种克隆的反刍动物反刍动物DNA探针与八种心水菌株杂交并检测出感染的绵羊。

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The DNA probe pCS20, which was cloned from the DNA of the Crystal Springs heartwater strain from Zimbabwe, cross-reacted with DNAs of heartwater strains from all endemic areas, including four heartwater strains from Zimbabwe, two strains from South Africa, one strain from Nigeria, and the Gardel strain from the Caribbean island of Guadeloupe. By nucleic acid hybridization, the pCS20 DNA probe detected Cowdria ruminantium DNA in all DNA preparations made from plasma samples from infected sheep before and during the febrile reaction. Synthetic oligonucleotides were prepared for amplification of specific C. ruminantium DNA sequences by the polymerase chain reaction (PCR). Amplification of two DNA products (181 and 279 bp) from pCS20 DNA and C. ruminantium genomic DNA of heartwater strains was demonstrated. In contrast, amplification of these products or any other products was not possible from genomic DNAs of Anaplasma marginale, Babesia bigemina, Trypanosoma brucei brucei, Escherichia coli, and bovine endothelial cells. The cross-reactivities of the 32P-labeled PCR products with genomic DNAs from several heartwater strains were similar to those with the pCS20 DNA probe. A nucleic acid-based test that uses hybridization assays and PCR provides a sensitive method for the detection of heartwater in both animals and ticks and has applications in epidemiological studies for the disease, which may allow for improved disease control.
机译：DNA探针pCS20是从津巴布韦的Crystal Springs心脏水株的DNA中克隆而来的，与来自所有地方性地区的心脏水株的DNA发生了交叉反应，包括来自津巴布韦的四株心脏水株，来自南非的两株，来自尼日利亚的一株。 ，以及来自加勒比海瓜德罗普岛的Gardel毒株。通过核酸杂交，pCS20 DNA探针在高热反应之前和过程中，检测了从受感染绵羊血浆样品制备的所有DNA制品中的反刍动物DNA。制备了用于通过聚合酶链反应（PCR）扩增特定反刍类反刍动物DNA序列的合成寡核苷酸。证明了从心水菌株的pCS20 DNA和反刍梭状芽胞杆菌基因组DNA扩增了两个DNA产物（181和279 bp）。相比之下，这些产物或任何其他产物的扩增不可能来自无缘无浆体，大巴贝虫，布鲁氏锥虫，布鲁氏菌，大肠杆菌和牛内皮细胞的基因组DNA。 32P标记的PCR产物与几种心水菌株的基因组DNA的交叉反应性与pCS20 DNA探针的交叉反应性相似。使用杂交测定和PCR的基于核酸的测试为检测动物和and中的心水提供了一种灵敏的方法，并且已在该疾病的流行病学研究中得到应用，这可能有助于改善疾病控制。

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