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首页> 外文期刊>Tropical biomedicine. >Development and validation of TaqMan real-time PCR for the detection of Burkholderia pseudomallei isolates from Malaysia
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Development and validation of TaqMan real-time PCR for the detection of Burkholderia pseudomallei isolates from Malaysia

机译：TaqMan实时PCR的开发和验证，用于检测马来西亚的伯克霍尔德菌假苹果毒株

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Rapid detection of Burkholderia pseudomallei, the etiologic agent of melioidosis,allows for timely initiation of appropriate treatment and better clinical outcomes. In thecurrent gold standard, the culture method is time consuming and suffers from low sensitivity.Meanwhile, previously reported molecular assays are fast and sensitive, but their performanceon isolates from Malaysia, an endemic region of melioidosis is under reported. This studydesigned oligonucleotides targeting orf2 of Type III secretion system (TTSS) genes clusterfor the detection of Malaysian B. pseudomallei isolates and evaluated the assay on 95 localB. pseudomallei strains, 58 other microorganisms and 71 clinical specimens from patients.The developed assay exclusively detected all tested B. pseudomallei isolates with a detectionlimit of 20 fg per reaction (equivalent to ~2.5 copies). Subsequent testing on clinical samplesshowed that the assay detected all confirmed specimens with the growth of B. pseudomallei(n = 10/10). None of the negative specimens had a detectable signal of our TTSS-orf2 assay(n = 0/61). In conclusion, the present study provides crucial preliminary data for a subsequentstudy and should be considered as a potential alternative to current time-consuming culturemethod for the detection of B. pseudomallei.
机译：快速检测类鼻疽病的病原体伯克霍尔德氏菌，可以及时开始适当的治疗并获得更好的临床效果。在当前的金标准中，该培养方法耗时且灵敏度低。同时，先前报道的分子测定是快速且灵敏的，但是其在来自马来西亚的类li虫病流行地区的分离株中的性能尚有报道。这项研究设计了针对III型分泌系统（TTSS）基因orf2的寡核苷酸簇，用于检测马来西亚假单胞菌分离株，并在95个localB上评估了该检测方法。假苹果芽孢杆菌菌株，58种其他微生物和71例来自患者的临床标本。开发的测定法仅检测所有测试的假芽孢杆菌菌株，每个反应的检出限为20 fg（约〜2.5份）。随后对临床样品进行的测试表明，该检测方法检测到所有确认的标本均带有假芽孢杆菌的生长（n = 10/10）。阴性样品均未检测到我们的TTSS-orf2检测信号（n = 0/61）。综上所述，本研究为后续研究提供了重要的初步数据，应被认为是目前检测假苹果芽孢杆菌耗时的培养方法的潜在选择。

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