Willems H, Thiele D, Krauss H
Institut für Hygiene und Infektionskrankheiten der Tiere, Justus-Liebig-Universität, Giessen, F.R.G.
Eur J Epidemiol. 1993 Jul;9(4):411-8. doi: 10.1007/BF00157399.
A "nested" PCR approach with primers based on conserved plasmid sequences was used for the highly sensitive and specific detection of Coxiella (C.) burnetii in clinical samples collected from cattle, dogs, cats and humans. Results were in good agreement with those obtained from Capture-ELISA and isolation of the organism in BGM cell culture. We also tested primers with sequences derived from genomic DNA and sequences based on 16S rRNA. In addition, we applied PCR for the differentiation of C. burnetii plasmid types from 28 isolates originating from the USA, Europe and South Africa. Reference isolates Nine Mile RSA493, Dugway 5J108-111 and all European isolates tested were recognized only by primers specific for the QpH1 plasmid. One isolate from a goat abortion in Namibia reacted identically to the reference isolate Priscilla Q177 bearing the QpRS plasmid. Reference isolate S Q217 with plasmid sequences integrated into the genome reacted with none of the plasmid-specific primer pairs.
采用基于保守质粒序列的引物进行“巢式”PCR方法，用于从牛、狗、猫和人类采集的临床样本中高度灵敏且特异检测伯氏考克斯体（C. burnetii）。结果与捕获ELISA以及在BGM细胞培养中分离该病原体所获得的结果高度一致。我们还测试了源自基因组DNA序列和基于16S rRNA序列的引物。此外，我们应用PCR对来自美国、欧洲和南非的28株分离株的伯氏考克斯体质粒类型进行区分。参考菌株九英里RSA493、达格威5J108 - 111以及所有测试的欧洲分离株仅被QpH1质粒特异性引物识别。纳米比亚一只山羊流产的一株分离株与携带QpRS质粒的参考菌株普里西拉Q177反应相同。质粒序列整合到基因组中的参考菌株S Q217与任何质粒特异性引物对均无反应。
