Deteksi Bakteri Patogen Salmonella typhi pada Sayuran yang dikonsumsi Mentah Menggunakan Metode nested Polymerase Chain Reaction

Idar Idar, Shinta Kusumawardhani, Mia Tria Novianti
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Abstract

Salmonella typhi (S. typhi) infection is a zoonotic infection and known as salmonellosis. In the human body, salmonellosis causes an increase in high body temperature or known as typhoid fever that cause high morbidity and mortality in developing countries, especially Indonesia. It was found that out of 22 million cases of typhoid fever, 200 thousand of them ended in death. S. typhi often contaminates food that was consumed raw or not perfectly cooked, for example meat, eggs, dairy products, fruits and vegetables. The conventional method for detecting these bacteria is culture method which time consuming and need BSL 2  facilities. PCR was one of DNA based detection method that could overcome the culture method weakness. In this study conducted detection of Salmonella bacteria in raw vegetables which are usually consumed as fresh by using nested PCR method. The detection procedures were sample preparation; bacterial DNA isolation; amplification by using two sets of primer, ST1-ST2 in first round PCR and ST3-ST4 in second round PCR; and the characterization by using agarose electrophoresis. The results indicated that two of nine raw vegetables, tomatos and cabbages have been contaminated with Salmonella. We conclude that nested PCR could detect Salmonella contamination in raw vegetables.
通过使用nested Polymerase链反应方法对生吃食用的蔬菜进行检测的沙门氏菌病原体
伤寒沙门氏菌感染是一种人畜共患感染,被称为沙门氏菌病。在人体内,沙门氏菌病导致体温升高,或称为伤寒,在发展中国家,特别是印度尼西亚,造成高发病率和死亡率。据调查,在2200万伤寒病例中,有20万人死亡。伤寒沙门氏菌通常会污染生食或未煮熟的食物,例如肉、蛋、乳制品、水果和蔬菜。传统的细菌检测方法是培养法,耗时长,需要BSL 2设备。PCR是一种基于DNA的检测方法,可以克服培养法的缺点。本研究采用巢式PCR法对新鲜食用的生蔬菜中的沙门氏菌进行了检测。检测程序为样品制备;细菌DNA分离;采用两组引物进行扩增,第一轮PCR为ST1-ST2,第二轮PCR为ST3-ST4;并用琼脂糖电泳进行表征。结果显示,9种生蔬菜、西红柿和卷心菜中有2种被沙门氏菌污染。结果表明,巢式PCR可以检测出蔬菜中沙门氏菌的污染情况。
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