Mohd Ali M R, Foo P C, Hassan M, Maning N, Hussin A, Syed Ahmad Yunus S Z, Fauzi M H, Muhd Besar A, Harun A, Ismail N, Chan Y Y
Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia.
Bacteriology Unit, Infectious Disease Research Center (IDRC), Institute for Medical Research (IMR), Complex National Institutes of Health (NIH), Section U13 Setia Alam, 40170 Shah Alam, Selangor, Malaysia.
Trop Biomed. 2019 Jun 1;36(2):379-389.
Rapid detection of Burkholderia pseudomallei, the etiologic agent of melioidosis, allows for timely initiation of appropriate treatment and better clinical outcomes. In the current gold standard, the culture method is time consuming and suffers from low sensitivity. Meanwhile, previously reported molecular assays are fast and sensitive, but their performance on isolates from Malaysia, an endemic region of melioidosis is under reported. This study designed oligonucleotides targeting orf2 of Type III secretion system (TTSS) genes cluster for the detection of Malaysian B. pseudomallei isolates and evaluated the assay on 95 local B. pseudomallei strains, 58 other microorganisms and 71 clinical specimens from patients. The developed assay exclusively detected all tested B. pseudomallei isolates with a detection limit of 20 fg per reaction (equivalent to ~2.5 copies). Subsequent testing on clinical samples showed 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 subsequent study and should be considered as a potential alternative to current time-consuming culture method for the detection of B. pseudomallei.
快速检测类鼻疽的病原体——伯克霍尔德菌，有助于及时开始适当治疗并取得更好的临床结果。在当前的金标准中，培养方法耗时且灵敏度低。同时，先前报道的分子检测方法快速且灵敏，但它们对来自类鼻疽流行地区马来西亚的分离株的检测性能报道较少。本研究设计了针对III型分泌系统（TTSS）基因簇的orf2的寡核苷酸，用于检测马来西亚伯克霍尔德菌分离株，并对95株本地伯克霍尔德菌菌株、58种其他微生物和71份患者临床标本进行了检测。所开发的检测方法仅能检测所有测试的伯克霍尔德菌分离株，检测限为每个反应20 fg（相当于约2.5个拷贝）。随后对临床样本的检测表明，该检测方法能检测出所有经培养确认有伯克霍尔德菌生长的标本（n = 10/10）。在阴性标本中，无一例在我们的TTSS-orf2检测中出现可检测信号（n = 0/61）。总之，本研究为后续研究提供了关键的初步数据，应被视为当前耗时的培养方法检测伯克霍尔德菌的潜在替代方法。
