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Potential use of serum based quantitative real-time PCR for the detection of pneumonia pathogens in a densely colonised population

机译：潜在的基于血清的实时荧光定量PCR检测人群密集的人群中的肺炎病原体

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Molecular methods offer improvement in the detection of causative pneumonia pathogens, but there are concerns of false positive results. Here we validate quantitative real-time PCR (qPCR) assays for the detection of Streptococcus pneumoniae and Haemophilus influenzae in: (a) spiked serum samples and (b) in matched serum and nasopharyngeal swabs from a population of Indigenous Australian children without pneumonia, but with a high nasopharyngeal carriage prevalence of S. pneumoniae and H. influenzae . Matched sera and nasopharyngeal swabs were selected from Indigenous children less than 5 years of age without a diagnosis of pneumonia. Specimens were assayed by qPCR targeting the lytA and glpQ genes from S. pneumoniae and H. influenzae , respectively. Using qPCR, neither S. pneumoniae nor H. influenzae DNA was detected in serum samples, even after concentration of serum DNA. In matched nasopharyngeal swabs, bacterial load was high with up to 106 cells/ml detected by qPCR. In this cohort of children with a high nasopharyngeal carriage, prevalence and bacterial load of pneumonia pathogens, qPCR on sera would not have produced a false pneumonia diagnosis. Thus, qPCR analysis of sera appears to be an appropriate method to aid aetiological diagnosis of pneumonia in this population.
机译：分子方法可改善对致病性肺炎病原体的检测，但存在假阳性结果的问题。在这里，我们验证了定量实时PCR（qPCR）检测方法在以下几个方面的检测：肺炎链球菌和流感嗜血杆菌的检测：（a）加标的血清样品，以及（b）来自无肺炎的澳大利亚土著儿童群体的匹配血清和鼻咽拭子鼻咽癌携带率高的肺炎链球菌和流感嗜血杆菌。配对的血清和鼻咽拭子选自5岁以下的未诊断为肺炎的土著儿童。通过针对分别来自肺炎链球菌和流感嗜血杆菌的lytA和glpQ基因的qPCR测定标本。使用qPCR，即使在浓缩血清DNA之后，在血清样品中也未检测到肺炎链球菌和流感嗜血杆菌DNA。在相配的鼻咽拭子中，细菌载量高，qPCR检测到高达106细胞/ ml。在该人群中，鼻咽癌携带率高，肺炎病原菌的患病率和细菌载量，血清中的qPCR不会产生错误的肺炎诊断。因此，对患者进行qPCR血清分析似乎是一种合适的方法，可帮助其对肺炎进行病因诊断。

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