Development and evaluation of Chlamylege, a new commercial test allowing simultaneous detection and identification of Legionella, Chlamydophila pneumoniae, and Mycoplasma pneumoniae in clinical respiratory specimens by multiplex PCR
Development and evaluation of Chlamylege, a new commercial test allowing simultaneous detection and identification of Legionella, Chlamydophila pneumoniae, and Mycoplasma pneumoniae in clinical respiratory specimens by multiplex PCR
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DOI：
10.1128/jcm.43.7.3247-3254.2005
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发表时间：
2005-07-01
影响因子：
9.4
通讯作者：
Grattard, F
中科院分区：
文献类型：
--
作者：
Ginevra, C;Barranger, C;Grattard, F
本研究描述了一种新的商业检测方法Chlamyge(Argene Inc.)的开发和评价，该方法允许通过多重聚合酶链式反应和微孔板杂交在呼吸道样本中同时检测肺炎衣原体、肺炎支原体和大多数军团菌以及聚合酶链式反应抑制剂。肺炎衣原体、肺炎支原体和嗜肺军团菌的敏感度分别为每管1×10(-3)IFU、5×10(-2)变色单位和1 CFU。用该试剂盒对154份呼吸道感染患者的临床标本进行了分析，其中肺炎衣原体感染2份，肺炎支原体感染9份，军团菌感染19份，三种病原体检测均为阴性114份。所有阳性标本均经衣原体试剂盒正确检测和鉴定，阴性标本未见假阳性结果。该试剂盒随后在一项儿科前瞻性研究中进行了评估，该研究包括220个气管内吸入物，并将结果与三种单一的室内PCR检测结果进行了比较。使用这两种方法，发现4个标本肺炎衣原体阳性，6个标本肺炎衣原体阳性。衣原体试剂盒检测到另外两个样本对肺炎支原体呈阳性，另一个样本对嗜肺乳杆菌以外的军团菌呈阳性；其他技术表明这三个样本为真阳性。这些结果表明，衣原体试验是敏感、特异和方便的，可以快速检测和鉴定呼吸道感染患者临床样本中的非典型病原体。
This study describes the development and evaluation of a new commercial test, Chlamylege (Argene Inc.), which allows the simultaneous detection in respiratory samples of Chlamydophila pneumoniae, Mycoplasma pneumoniae, and most Legionella species, as well as PCR inhibitors, by using a multiplex PCR and microplate hybridization. The sensitivities of Chlamylege were 1 x 10(-3) IFU, 5 x 10(-2) color-changing units, and 1 CFU per reaction tube for C. pneumoniae, M. pneumoniae, and Legionella pneumophila, respectively. A cohort of 154 clinical samples from patients with documented respiratory infections was analyzed by the kit, including 2 samples from patients with C. pneumoniae infection, 9 samples from patients with M. pneumoniae infection, 19 samples from patients with Legionella species infection, and 114 samples that tested negative for the three pathogens. All the positive specimens were correctly detected and identified by the Chlamylege kit, and no false-positive result was observed with the negative samples. The kit was then evaluated in a pediatric prospective study that included 220 endotracheal aspirates, and the results were compared with those obtained by three single in-house PCR assays. Four specimens were found to be positive for C. pneumoniae and six were found to be positive for M. pneumoniae by using both strategies. The Chlamylege kit detected two additional samples positive for M. pneumoniae and one additional sample positive for a Legionella species other than L. pneumophila; these three samples were shown to be true positive by other techniques. These overall results demonstrate that the Chlamylege assay is sensitive, specific, and convenient for the rapid detection and identification of atypical pathogens in clinical samples from patients with respiratory infections.