Pui Chai Fung, Wong Woan Chwen, Chai Lay Ching, Lee Hai Yen, Noorlis Ahmad, Zainazor Tuan Chilek Tuan, Tang John Yew Huat, Ghazali Farinazleen Mohamad, Cheah Yoke Kqueen, Nakaguchi Yoshitsugu, Nishibuchi Mitsuaki, Radu Son
Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia.
Trop Med Health. 2011 Mar;39(1):9-15. doi: 10.2149/tmh.2010-20. Epub 2011 Mar 24.
Salmonellosis outbreaks involving typhoid fever and human gastroenteritis are important diseases in tropical countries where hygienic conditions are often not maintained. A rapid and sensitive method to detect Salmonella spp., Salmonella Typhi and Salmonella Typhimurium is needed to improve control and surveillance of typhoid fever and Salmonella gastroenteritis. Our objective was the concurrent detection and differentiation of these food-borne pathogens using a multiplex PCR. We therefore designed and optimized a multiplex PCR using three specific PCR primer pairs for the simultaneous detection of these pathogens. The concentration of each of the primer pairs, magnesium chloride concentration, and primer annealing temperature were optimized before verification of the specificity of the primer pairs. The target genes produced amplicons at 429 bp, 300 bp and 620 bp which were shown to be 100% specific to each target bacterium, Salmonella spp., Salmonella Typhi and Salmonella Typhimurium, respectively.
在卫生条件常常难以维持的热带国家，涉及伤寒热和人类肠胃炎的沙门氏菌病疫情是重要的疾病。需要一种快速且灵敏的方法来检测沙门氏菌属、伤寒沙门氏菌和鼠伤寒沙门氏菌，以加强对伤寒热和沙门氏菌肠胃炎的控制与监测。我们的目标是使用多重聚合酶链反应（multiplex PCR）同时检测和区分这些食源性病原体。因此，我们设计并优化了一种多重聚合酶链反应，使用三对特异性PCR引物来同时检测这些病原体。在验证引物对的特异性之前，对每对引物的浓度、氯化镁浓度和引物退火温度进行了优化。目标基因产生了分别为429 bp、300 bp和620 bp的扩增子，结果表明这些扩增子对每种目标细菌，即沙门氏菌属、伤寒沙门氏菌和鼠伤寒沙门氏菌分别具有100%的特异性。
