The Relevance of a Novel Quantitative Assay to Detect up to 40 Major Streptococcus pneumoniae Serotypes Directly in Clinical Nasopharyngeal and Blood Specimens.
The Relevance of a Novel Quantitative Assay to Detect up to 40 Major Streptococcus pneumoniae Serotypes Directly in Clinical Nasopharyngeal and Blood Specimens.
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DOI：
10.1371/journal.pone.0151428
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Telles JN
中科院分区：
文献类型：
--
作者：
Messaoudi M;Milenkov M;Albrich WC;van der Linden MP;Bénet T;Chou M;Sylla M;Barreto Costa P;Richard N;Klugman KP;Endtz HP;Paranhos-Baccalà G;Telles JN
为了流行病学和监测的目的，监测肺炎链球菌血清型的分布和动态是相关的。传统的血清分型方法不能提供有关血清型载量的快速或定量信息。定量血清分型可以预测特定血清型与携带的其他血清型相比的侵袭性。在这里，我们描述了一种新型、快速的多重实时PCR检测方法，用于识别和定量40种最流行的肺炎球菌血清型，以及该检测方法对新兴国家和发展中国家肺炎样本的影响。优化了11条多重PCR检测40个血清型或血清群的方法。通过参考已知细菌负荷的标准稀释液进行定量。评价了检测试剂盒的性能，以特异性分型和定量S。来自五个不同国家的成人和儿科肺炎住院患者（n = 664）的鼻咽和血液样本中的肺炎。血清群6在所有5个队列的鼻咽标本中广泛存在。法国、南非和巴西队列中最常见的血清型分别为1和7A/F、3和19 F以及14。当两种样本都可用时，血液中的血清型总是与鼻咽中的其他血清型一起携带。此外，血清型侵入血流的能力可能与其鼻咽负荷有关。移动到血液中的血清型的平均鼻咽浓度比仅在鼻咽中发现的血清型高3个对数倍。这种新的、快速的、定量的检测方法可能潜在地预测一些S。肺炎血清型侵袭性和肺炎球菌血清型分布的评估。
For epidemiological and surveillance purposes, it is relevant to monitor the distribution and dynamics of Streptococcus pneumoniae serotypes. Conventional serotyping methods do not provide rapid or quantitative information on serotype loads. Quantitative serotyping may enable prediction of the invasiveness of a specific serotype compared to other serotypes carried. Here, we describe a novel, rapid multiplex real-time PCR assay for identification and quantification of the 40 most prevalent pneumococcal serotypes and the assay impacts in pneumonia specimens from emerging and developing countries. Eleven multiplex PCR to detect 40 serotypes or serogroups were optimized. Quantification was enabled by reference to standard dilutions of known bacterial load. Performance of the assay was evaluated to specifically type and quantify S. pneumoniae in nasopharyngeal and blood samples from adult and pediatric patients hospitalized with pneumonia (n = 664) from five different countries. Serogroup 6 was widely represented in nasopharyngeal specimens from all five cohorts. The most frequent serotypes in the French, South African, and Brazilian cohorts were 1 and 7A/F, 3 and 19F, and 14, respectively. When both samples were available, the serotype in blood was always present as carriage with other serotypes in the nasopharynx. Moreover, the ability of a serotype to invade the bloodstream may be linked to its nasopharyngeal load. The mean nasopharyngeal concentration of the serotypes that moved to the blood was 3 log-fold higher than the ones only found in the nasopharynx. This novel, rapid, quantitative assay may potentially predict some of the S. pneumoniae serotypes invasiveness and assessment of pneumococcal serotype distribution.