A highly sensitive and specific real-time PCR assay for the detection of spotted fever and typhus group rickettsiae
A highly sensitive and specific real-time PCR assay for the detection of spotted fever and typhus group rickettsiae
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DOI：
10.4269/ajtmh.2005.73.1083
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发表时间：
2005-12-01
影响因子：
3.3
通讯作者：
Unsworth, NB
中科院分区：
文献类型：
--
作者：
Stenos, J;Graves, SR;Unsworth, NB
以柠檬酸合成酶基因为靶点，建立了检测斑点热立克次体和斑疹伤寒群立克次体的高特异性实时聚合酶链式反应(PCR)方法。该方法扩增了斑点热群和斑疹伤寒群的立克次体成员，包括阿卡里立克次体、澳大利亚立克次体、康诺里立克次体、霍尼立克次体、“马尔米尼立克次体”、西伯利亚立克次体、立克次体、伤寒立克次体和普氏立克次体。贝氏立克次体的祖先群没有产生阳性反应，立克次体目的其他成员或任何非立克次体细菌也没有产生阳性反应。该方法的灵敏度为每次反应一个靶拷贝数，由含有斑点热群靶序列的质粒的连续稀释确定。当立克次体数量很低时，这种定量检测方法可用于临床标本中立克次体的计数和立克次体疾病的诊断。
A highly specific real-time polymerase chain reaction (PCR) assay was developed to detect spotted fever and typhus group rickettsiae using the citrate synthase gene as the target. The assay amplified rickettsial members of the spotted fever and typhus group including Rickettsia akari, R. australis, R. conorii, R. honei, "R. marmionii, " R. sibirica, R. rickettsii, R. typhi, and R. prowazekii. The ancestral group rickettsia, R. bellii, did not produce a positive reaction, nor did other members of the order Rickettsiales or any non-rickettsial bacteria. The assay had a sensitivity of one target copy number per reaction as determined by serial dilutions of a plasmid containing a spotted fever group target sequence. This quantitative assay is useful for the enumeration of rickettsiae in clinical specimens and the diagnosis of rickettsial illnesses, when rickettsial numbers are very low.