Characterization of a duplex PCR assay for the detection of enterotoxigenic strains of Staphylococcus aureus

作     者：Junni Tang~(1,2) Xianming Shi Chunlei Shi~1 Huanchun Chen~2 (1Department of Food Science and Technology and Bor Luh Food Safety Center,Shanghai Jiao Tong University,Shanghai 201101 China)(2National Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070 China) 

会议名称：《中国食品科学技术学会第五届年会暨第四届东西方食品业高层论坛》

会议日期：2007年

学科分类：09[农学] 0903[农学-农业资源与环境] 

关 键 词：duplex PCR uniform design Staphylococcus aureus rapid detection 

摘      要：Sequence analysis demonstrated that all reported primers based on the thermonuclease gene (nuc)used in the detection of the food-borne pathogen,Staphylococcus aureus,were derived from the nuclease gene,instead of the thermonuclease *** this study,two pairs of primers targeting the sequences of the nuc gene and 16S rRNA gene were designed in a duplex PCR assay for the detection of ***,particularly those of the enterotoxigenic *** optimization of the amplification conditions by uniform design,two specific products were amplified in the PCR detection system:a 223-bp nuc fragment and a 565-bp 16S rDNA *** tests demonstrated that this duplex-PCR method has high specificity for only *** detection limit of the assay was 9.35 pg/μl using *** genomic *** sensitivity for direct detection of milk samples sparked with S. aureus was 104 to 105 CFU/ml,and the sensitivity was 1 CFU/ml as an initial concentration when an enrichment process was used.A PCR-based detection assay for enterotoxin gene was performed in parallel with the detection of the nuc-encoded TNase activity in several *** strains,and a good correlation between TNase production and enterotoxigenicity was demonstrated in all the strains tested.