Evaluation of different nucleic acid amplification techniques for the detection of M-pneumoniae, C-pneumoniae and Legionella spp. in respiratory specimens from patients with community-acquired pneumonia
Evaluation of different nucleic acid amplification techniques for the detection of M-pneumoniae, C-pneumoniae and Legionella spp. in respiratory specimens from patients with community-acquired pneumonia
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DOI：
10.1016/j.mimet.2008.02.010
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发表时间：
2008-06-01
影响因子：
2.2
通讯作者：
Leven, M.
中科院分区：
文献类型：
--
作者：
Loens, K.;Beck, T.;Leven, M.
社区获得性肺炎涉及的病原体数量众多，对抗菌剂的敏感性各不相同，这对诊断微生物学构成了巨大挑战。肺炎链球菌感染与肺炎支原体、肺炎嗜衣原体或L.比较了实时单纯和多重核酸序列扩增（NASBA）和单纯PCR检测嗜肺支原体的敏感性和特异性。pneumoniae、C.肺炎和军团菌属。对147例社区获得性肺炎（CAP）患者的251份呼吸道标本进行了检测。使用NucliSens Basic试剂盒（bioMerieux）进行NASBA。PCR检测M. pneumoniae和C.如前所述[Leven，M.，Ursi，D.，货车贝弗，H.，昆特，西，尼斯特斯，H. G. M.，和Goossens，H. 1996.两种聚合酶链反应检测肺炎支原体的研究。肺炎在儿科患者急性呼吸道感染中的作用。J. Infect. Dis.173，1445 -14452. Ursi，D.，Leven，M.，货车贝弗，H. P.，和Goossens，H. 1998.肺炎衣原体PCR检测内参的构建摩尔Cellul Probes. 12，235-238.]。建立了实时荧光定量PCR检测L.而实时NASBA被设计用于检测军团菌属。对所有不一致结果的样品进行重新分析，与扩展金标准相比，不同技术检测M的灵敏度分别为77.8%、100%和100%。对肺炎克雷伯氏菌的检出率分别为50%、100%和50%。单纯实时NASBA被证明比单纯PCR更敏感，并且也比实时多重NASBA更敏感，如先前在加标临床标本中发现的。它的实际吸引力恳求进一步优化的多重方法。(C)2008 Elsevier B. V.保留所有权利。
The number of pathogens involved in community-acquired pneumonia, with varying susceptibilities to antimicrobials, is numerous constituting an enormous challenge for diagnostic microbiology. Differentiation of infections due to Streptococcus pneumoniae and those due to Mycoplasma pneumoniae, Chlamydophila pneumoniae, or L. pneumophila as well as those due to viruses is essential to allow correct decisions concerning the antibiotics to be administered.The sensitivity and specificity of real-time simplex and multiplex nucleic acid sequence-based amplification (NASBA), and simplex PCR were compared for the detection of M. pneumoniae, C. pneumoniae and Legionella spp. in respiratory specimens from hospitalized and outpatients with community-acquired pneumonia (CAP).Two hundred fifty one respiratory specimens were collected from 147 patients with CAP. NASBA was done using the NucliSens Basic Kit (bioMerieux). PCR for M. pneumoniae and C. pneumoniae was done as described earlier [leven, M., Ursi, D., Van Bever, H., Quint, W., Niesters, H. G. M., and Goossens, H. 1996. Detection of Mycoplasma pneumoniae by two polymerase chain reactions and role of M. pneumoniae in acute respiratory tract infections in pediatric patients. J. Infect. Dis. 173,1445-14452.; Ursi, D., leven, M., Van Bever, H. P., and Goossens, H. 1998. Construction of an internal control for the detection of Chlamydia pneumoniae by PCR. Mol. Cellul. Probes. 12, 235-238.]. A real-time PCR was developed to detect L. pneumophila whereas a real-time NASBA was designed to detect Legionella spp. All samples with discordant results were re-analysed.Compared to an expanded gold standard the sensitivities of the different techniques, were 77.8%, 100%, and 100% for detection of M. pneumoniae; and 50%, 100%, and 50% for detection of L pneumophila by PCR, real-time simplex NASBA, and real-time multiplex NASBA, respectively.C. pneumoniae was detected in two samples only.Simplex real-time NASBA proved to be more sensitive than simplex PCR and was also more sensitive than real-time multiplex NASBA, as previously found with spiked clinical specimens. It's practical attractiveness pleads for further optimalisation of the multiplex approach. (C) 2008 Elsevier B.V. All rights reserved.