Abdullah J, Saffie N, Sjasri F A R, Husin A, Abdul-Rahman Z, Ismail A, Aziah I, Mohamed M
Institute for Research in Molecular Medicine Kubang Kerian Kelantan Institute for Research in Molecular Medicine, Kubang Kerian, Kelantan.
Hospital Raja Perempuan Zainab II 15586 Kota BharuKelantan Malaysia Hospital Raja Perempuan Zainab II, 15586 Kota Bharu, Kelantan, Malaysia.
Braz J Microbiol. 2015 Mar 4;45(4):1385-91. doi: 10.1590/s1517-83822014000400032. eCollection 2014.
An in-house loop-mediated isothermal amplification (LAMP) reaction was established and evaluated for sensitivity and specificity in detecting the presence of Salmonella Typhi (S. Typhi) isolates from Kelantan, Malaysia. Three sets of primers consisting of two outer and 4 inner were designed based on locus STBHUCCB_38510 of chaperone PapD of S. Typhi genes. The reaction was optimised using genomic DNA of S. Typhi ATCC7251 as the template. The products were visualised directly by colour changes of the reaction. Positive results were indicated by green fluorescence and negative by orange colour. The test was further evaluated for specificity, sensitivity and application on field samples. The results were compared with those obtained by gold standard culture method and Polymerase Chain Reaction (PCR). This method was highly specific and -10 times more sensitive in detecting S. Typhi compared to the optimised conventional polymerase chain reaction (PCR) method.
建立了一种内部环介导等温扩增（LAMP）反应，并对其检测来自马来西亚吉兰丹伤寒沙门氏菌（伤寒杆菌）分离株的敏感性和特异性进行了评估。基于伤寒杆菌基因伴侣蛋白PapD的基因座STBHUCCB_38510设计了三组引物，包括两个外部引物和四个内部引物。以伤寒杆菌ATCC7251的基因组DNA为模板对反应进行优化。通过反应颜色变化直接观察产物。绿色荧光表示阳性结果，橙色表示阴性结果。对该检测方法的特异性、敏感性及在现场样本中的应用进行了进一步评估。将结果与通过金标准培养法和聚合酶链反应（PCR）获得的结果进行比较。与优化后的传统聚合酶链反应（PCR）方法相比，该方法检测伤寒杆菌具有高度特异性且灵敏度高10倍。
