Clinical Trials and Research Unit, Changi General Hospital, 2 Simei Street 3, Singapore 529889, Singapore.
Department of Laboratory Medicine, Changi General Hospital, 2 Simei Street 3, Singapore 529889, Singapore.
Trans R Soc Trop Med Hyg. 2021 Dec 2;115(12):1450-1455. doi: 10.1093/trstmh/trab079.
Community-acquired pneumonia (CAP) is one of the most common infectious diseases and is a significant cause of mortality and morbidity globally. A microbial cause was not determined in a sizable percentage of patients with CAP; there are increasing data to suggest regional differences in bacterial aetiology. We devised a multiplex real-time PCR assay for detecting four microorganisms (Streptococcus pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae and Burkholderia pseudomallei) of relevance to CAP infections in Asia.
Analytical validation was accomplished using bacterial isolates (n=10-33 of each target organism for analytical sensitivity and n=117 for analytical sensitivity) and clinical validation using 58 culture-positive respiratory tract specimens.
The qPCR assay exhibited 100% analytical sensitivity and analytical specificity, and 100% clinical sensitivity and 94-100% clinical specificity. The limit of detection and efficiency for the multiplex PCR assay were 3-33 CFU/mL and 93-110%, respectively. The results showed that the PCR-based method had higher sensitivity than traditional culture-based methods. The assay also demonstrated an ability to semiquantify bacterial loads.
We have devised a reliable laboratory-developed multiplex qPCR assay, with a turnaround time of within one working day, for detection of four clinically important CAP-associated microorganisms in Asia. The availability of a test with improved diagnostic capabilities potentially leads to an informed choice of antibiotic usage and appropriate management of the patient to achieve a better treatment outcome and financial savings.
社区获得性肺炎（CAP）是最常见的传染病之一，也是全球范围内导致死亡率和发病率的主要原因。相当一部分 CAP 患者的病因无法确定微生物；越来越多的数据表明，细菌病因在不同地区存在差异。我们设计了一种多重实时 PCR 检测方法，用于检测与亚洲 CAP 感染相关的四种微生物（肺炎链球菌、流感嗜血杆菌、肺炎克雷伯菌和鼻疽伯克霍尔德菌）。
采用细菌分离株（每种目标生物的分析灵敏度为 10-33 个，分析灵敏度为 117 个）进行分析验证，采用 58 份培养阳性呼吸道标本进行临床验证。
qPCR 检测方法具有 100%的分析灵敏度和分析特异性，100%的临床灵敏度和 94-100%的临床特异性。该多重 PCR 检测方法的检测限和效率分别为 3-33 CFU/mL 和 93-110%。结果表明，基于 PCR 的方法比传统的基于培养的方法具有更高的灵敏度。该检测方法还能够半定量细菌负荷。
我们已经开发出一种可靠的实验室开发的多重 qPCR 检测方法，其周转时间在一个工作日内，用于检测亚洲四种与 CAP 相关的重要临床微生物。具有改进诊断能力的检测方法的出现，可能会导致更明智地选择抗生素使用，并对患者进行适当的管理，以实现更好的治疗效果和节省成本。
