摘要
为建立溶藻弧菌(VA)的快速检测方法,本研究以VA Collagenase基因为靶基因设计合成引物及Taq Man探针,建立了实时荧光定量PCR快速检测VA的方法。结果显示,对15株实验菌株进行荧光定量PCR检测,只有VA菌株检测为阳性,表明该检测方法特异性强;该方法的灵敏度为18 cfu/m L;稳定性和重复性实验结果表明,同一样品重复检测4次Ct值的变异系数均小于2%;利用该检测方法对采集的165份样品进行检测,共计检出2份VA阳性样品,与SN/T2564-2010行标法检测结果一致,显示了良好的实用性。该检测方法灵敏度高、特异性强,具有良好的实用性。
To establish a rapid assay for Vibrio alginolyticus（VA） detection,a dual real-time PCR method was developed targeting Collagenase gene of VA. The results showed that the test for 15 bacteria strains of the dual realtime PCR method, only VA test was positive, indicating that the method had high specificity. In addition,the sensitivity of a dual real-time PCR was 18 cfu/m L. Stability and reproducibility of the test results showed that the coefficient of variation of the same sample repeat the test four times Ct values were less than2%. Furthermore,a total 2 positive samples for VA were detected from 165 clinical samples by the real-time method,which was in accordance with the testing result by SN/T 2564-2010 standard detection protocol.Therefore,the real-time method provides a novel rapid and sensitive detection method for VA infection.
出处
《食品工业科技》
CAS
CSCD
北大核心
2016年第8期69-71,76,共4页
Science and Technology of Food Industry
基金
海南省社会发展科技专项(2015SF29)
国家质检总局科技项目(2013IK031
2013IK051
2015IK089)
重庆市科技计划项目(cstc2014yykf A80017)
海南省应用技术研究与开发专项项目(ZDXM20130025)
广东检验检疫局科技计划项目(2011GDK44
2013GDK04)
