Position: Home > Articles > Research on Detection of Salmonella Combining EMA and PCR
Animal Husbandry and Feed Science
2013
(11)
113-116
EMA与PCR结合检测沙门氏菌方法的研究
作 者：
Yanan Zha
单 位：
Henan Animal Products Quality Monitor Test Cente
关键词：
ema;salmonella;concentration;amplification;target gene;exposure tim
摘 要：
为了建立一种快速区分样品中沙门氏菌的死细菌与活细菌的检测方法,将叠氮溴化乙锭(EMA)与PCR技术相结合,以沙门氏菌的invA为靶基因,以沙门氏菌的纯培养细胞做模板进行扩增,并进行灵敏度、特异性、曝光时间及EMA浓度试验。结果显示,灵敏度为14 CFU/mL,最佳曝光时间为2 min,当EMA的浓度小于18μg/mL时,EMA对活菌靶基因的扩增没有明显的抑制,而终浓度为1μg/mL的EMA,能有效抑制1×108CFU/mL沙门氏菌死菌的扩增。EMA-PCR能有效降低沙门氏菌检测过程中的假阳性。
译 名：
Research on Detection of Salmonella Combining EMA and PCR
作 者：
ZHAO Yu;LI Jin-lei;DONG Peng;DI Yuan-ran;GAO Yan-ling;Zhumadian Animal Products Quality Monitor Test Center of Henan Province;Henan Animal Products Quality Monitor Test Center;
关键词：
Salmonella;;EMA-PCR;;detection;;dead bacteria and live bacteria
摘 要：
In order to establish a fast detection method for distinguishing dead bacteria and live bacteria of Salmonella from samples, EMA and PCR were combined, meanwhile invA gene of Salmonella was taken as target gene, and then amplification was carried out by taking pure culture cells of Salmonella as template, furthermore sensitivity, specificity, exposure time and EMA concentration test were carried out. Results showed that: the sensitivity was 14 CFU/mL; the optimal exposure time was 2 min; when the concentration of EMA was less than 18 μg/mL, EMA had no obvious inhibition to the amplification of live bacteria′ target gene, while EMA at final concentration of 1 μg/mL could effectively inhibit the amplification of dead bacteria at 1×108CFU/mL. So EMA-PCR could effectively decrease false positive rate in the detection of Salmonella.
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