一种多重嵌套 PCR 方法，可同时检测来自印度南部 Vellore 的不明原因发热（PUO）患者中的伤寒沙门氏菌、结核分枝杆菌和类鼻疽伯克霍尔德菌。
A multiplex nested PCR for the simultaneous detection of Salmonella typhi, Mycobacterium tuberculosis, and Burkholderia pseudomallei in patients with pyrexia of unknown origin (PUO) in Vellore, South India.
机构信息
Sri Sakthi Amma Institute of Biomedical Research, Sri Narayani Hospital and Research Centre, Sripuram, Vellore, 632 055, Tamil Nadu, India,
出版信息
Mol Diagn Ther. 2014 Jun;18(3):315-21. doi: 10.1007/s40291-013-0079-7.
BACKGROUND AND OBJECTIVES
Salmonella typhi, Mycobacterium tuberculosis, and Burkholderia pseudomallei are among the most important monocyte-tropic bacterial agents causing pyrexia of unknown origin (PUO), with a significant number of endemic infections in both South and Southeast Asian regions. These infections pose a major risk to travelers to these regions as well.
METHODS
We developed and evaluated a multiplex nested polymerase chain reaction (PCR) for the simultaneous detection of the three pathogens in 305 patients' buffy coat samples.
RESULTS
The assay for S. typhi and B. pseudomallei was able to detect down to 1 colony forming unit/5 μL PCR input and M. tuberculosis was detected down to 20 genome copies/5 μL PCR input. S. typhi was detected in 10 (3.3 %) individuals, B. pseudomallei in 10 individuals (3.3 %), and M. tuberculosis in 18 individuals (5.9 %). Co-infections of M. tuberculosis and B. pseudomallei were detected in three individuals and S. typhi and B. pseudomallei in two individuals.
CONCLUSION
This protocol is efficient for PUO diagnosis especially in Asian countries.
背景和目的
伤寒沙门氏菌、结核分枝杆菌和类鼻疽伯克霍尔德菌是引起不明原因发热（PUO）的最重要的单核细胞亲代细菌病原体之一，在南亚和东南亚地区都有大量地方性感染。这些感染也对前往这些地区的旅行者构成重大威胁。
方法
我们开发并评估了一种多重巢式聚合酶链反应（PCR），用于同时检测 305 名患者的白细胞样本中的三种病原体。
结果
该检测方法可检测到伤寒沙门氏菌和类鼻疽伯克霍尔德菌的检测下限为 1 个菌落形成单位/5 μL PCR 输入，结核分枝杆菌的检测下限为 20 个基因组拷贝/5 μL PCR 输入。10 名个体（3.3%）中检测到伤寒沙门氏菌，10 名个体（3.3%）中检测到类鼻疽伯克霍尔德菌，18 名个体（5.9%）中检测到结核分枝杆菌。在 3 名个体中检测到结核分枝杆菌和类鼻疽伯克霍尔德菌的合并感染，在 2 名个体中检测到伤寒沙门氏菌和类鼻疽伯克霍尔德菌的合并感染。
结论
该方案对 PUO 诊断有效，特别是在亚洲国家。
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