药品中沙门菌荧光PCR法的建立

Establishment of Fluorescence PCR Method for the Detection of Salmonella in Drugs

建立药品中沙门菌检查的荧光PCR检测方法。梯度稀释沙门菌悬液确定该方法的灵敏度,设置沙门菌液组、大肠埃希菌菌液组、沙门菌及大肠埃希菌混合菌液组、空白对照组共4个试验组,考察该方法的专属性,选取5批样品分别设置供试品组和阳性对照组,考察该方法的重现性及耐用性。该方法检测灵敏度为103cfu/m L,4组试验中,沙门菌液组、沙门菌及大肠埃希菌混合菌液组均检出沙门菌,大肠埃希菌组及空白对照均未检出。5组样品中,采用PCR法和药典标准方法,两种方法检查结果符合率为100%。该方法灵敏度高,不受大肠埃希菌干扰,具有良好的专属性、重现性及耐用性,可以用于药品中沙门菌的检查。

To establish a fluorescence polymerase chain reaction( PCR) based method for the detection of Salmonella in drugs,serial dilutions of Salmonella suspension were used for the sensitivity test. Four experimental groups were established Salmonella suspension group,E. coli suspension group,E. coil and Salmonella suspension mixed group and blank control group for the specificity test. The reproducibility and durability test were investigated by 5 batches of representative samples. The method was much specific and not affected by E. coli interference,the detection limit was 103 cfu/m L. The Salmonella suspension group and the E. coil-Salmonella suspension mixed group were positive results while the E. coil suspension group and the blank control group were negative results.The detection rate was consistent with that of pharmacopoeia method. Good sensitivity、specificity and reproducibility were found in the study,PCR could be probably used in the detection of Salmonella in drugs.