Templeton Kate E, Scheltinga Sitha A, Sillekens Peter, Crielaard Jantine W, van Dam Alje P, Goossens Herman, Claas Eric C J
Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands.
J Clin Microbiol. 2003 Sep;41(9):4016-21. doi: 10.1128/JCM.41.9.4016-4021.2003.
A multiplex real-time PCR assay for detection of Legionella pneumophila and Legionella spp. and including an internal control was designed. Legionella species, L. pneumophila, and the internal control were detected simultaneously by probes labeled with 6-carboxy-fluorescein, hexachlorofluorescein, and indodicarbocyanine, respectively. Therefore, no postamplification analysis was required in order to distinguish the targets. The sensitivity of both assays was 2.5 CFU/ml, and from analysis of 10 culture-positive and 74 culture-negative samples from patients investigated for legionellosis, 100% agreement was observed by both assays in comparison to culture. Four additional positives were found by the multiplex real-time PCR assay in the Legionella culture-negative samples.
设计了一种用于检测嗜肺军团菌和军团菌属并包含内部对照的多重实时聚合酶链反应（PCR）检测方法。分别用6-羧基荧光素、六氯荧光素和吲哚二碳菁标记的探针同时检测军团菌属、嗜肺军团菌和内部对照。因此，无需进行扩增后分析来区分目标物。两种检测方法的灵敏度均为2.5 CFU/ml，在对10例军团病调查患者的培养阳性和74例培养阴性样本进行分析时，与培养结果相比，两种检测方法的一致性均为100%。在军团菌培养阴性样本中，多重实时PCR检测方法还发现了另外4个阳性结果。
