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[1]梁洪军,李 晶,王永山,等.应用荧光定量PCR检测食品中沙门菌[J].医学研究与战创伤救治(原医学研究生学报),2013,15(03):215-218.[doi:10.3969/j.issn.1672-271X.2013.03.003]
　LIANG Hong-jun,LI Jing,WANG Yong-shan,et al.Detecting Salmonella in food by fluorescence quantitative PCR technique[J].JOURNAL OF MEDICALRESEARCH —COMBAT TRAUMA CARE,2013,15(03):215-218.[doi:10.3969/j.issn.1672-271X.2013.03.003]
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应用荧光定量PCR检测食品中沙门菌()
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《医学研究与战创伤救治》（原医学研究生学报）[ISSN:1672-271X/CN:32-1713/R]

卷:
第15卷
期数:
2013年03期
页码:
215-218
栏目:
出版日期:
2013-03-30

文章信息/Info

Title:
Detecting Salmonella in food by fluorescence quantitative PCR technique
作者:
梁洪军1; 李 晶1; 王永山2; 魏德江1; 陈 琼1; 周东明1; 唐雨德1
1.210002 江苏南京,南京军区疾病预防控制中心; 2.210014 江苏南京,江苏省农业科学院兽医所
Author(s):
LIANG Hong-jun1; LI Jing1; WANG Yong-shan2; WEI De-jiang1; CHEN Qiong1; ZHOU Dong-ming1; Tang Yu-de1
1.Center for Disease Control and Prevention of Nanjing Military Command,Nanjing,Jiangsu 210002,China; 2.Institute of Veterinary,Jiangsu Province Academy of Agricultural Sciences,Nanjing,Jiangsu 210014,China
关键词:
沙门菌; 荧光定量PCR; 检测
Keywords:
Salmonella;  fluorescence quantitative PCR;  detecting
分类号:
R516.3
DOI:
10.3969/j.issn.1672-271X.2013.03.003
文献标志码:
A
摘要:
目的 应用荧光定量PCR技术检测食品中沙门菌,为病原监测和快速诊断提供实验依据。 方法 选择沙门菌invA基因设计引物,应用SYBRGreen I染料建立荧光定量PCR法。分别对61株沙门菌、14株非沙门菌以及食品模拟标本、市售禽蛋制品进行检测,观察方法的特异性、敏感性和可行性。结果 建立的荧光定量PCR法检测所有沙门菌均出现了特异的熔解曲线,扩增产物的熔点值79.6 ℃左右; 目标菌DNA的扩增产物循环数(Ct值)为20,空白对照及非沙门菌的Ct值大于30,非沙门菌均未出现特异的熔解曲线。每反应最低检测的沙门菌数是31 CFU; 对食品中人工污染的沙门菌的检测,最低检测量是1 CFU,检测时间为7～8 h,与培养法比较,阳性符合率100%; 对市售的369份禽蛋制品进行检测,阳性结果19份,而细菌培养法检测阳性结果为12份。结论 基于SYBR Green I染料建立的荧光定量PCR检测沙门菌是敏感、特异、省时、省力的方法,适用于沙门菌快速检测。
Abstract:
Objective To develop a fluorescence quantitative PCR technique for detecting Salmonella in food.Methods A pair of specific primers for Salmonella invA gene were screened.SYBRGreen I was used to establish the method of fluorescence quantitative polymerase chain reaction for detecting Salmonella.61 strains of Salmonella and 14 strains of other bacteria,food simulation specimens and poultry products were detected to test its sensitivity,specificity and possibility.Results All Salmonella appeared the specific melting curve with the developed fluorescence quantitative PCR,the melting point of the target DNA was about 79.6 ℃.The Ct value was about 20.The Ct value of blank control and other bacteria were greater than 30 and didn't appear the specific melting curve.The lowest detecting number was 31 CFU each reaction and 1 CFU in the artificial contamination food,respectively.Testing time was 7 to 8 hours,and its positive coincidence rate was 100% in comparison with culture method.Detecting 369 copies of the selling poultry products with the method,the positive number was 19,but 12 with bacteria culture method.Conclusion The developed fluorescence quantitative PCR array was sensitive,specific and efficient.The processing was rapid and simple.The method is suitable for rapid detection of Salmonella.

参考文献/References:

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备注/Memo

备注/Memo:
基金项目: 南京军区“十一五”医药卫生科研基金课题(06Z54); 南京军区卫生专业人才培养“122”工程资助项目
更新日期/Last Update: 2013-05-20