Abstract
The degenerate primers were designed to establish a PCR assay for synchronous and fast detection of staphylococcal enterotoxin A and B. According to the sequences of SEA and SEB gene, primers were designed and used for PCR. The lengths of the amplified primers were 105 bp and 135 bp. To evaluate the specificity of PCR assay, 2 strains of SEA, SEB and 4 strains of non-SEA-B were tested by PCR. SEB was 10-fold serially diluted and amplified by PCR to verify its sensitivity. The results showed that the detection of Staphylococcus aureus enterotoxin A, B strain DNA results were positive and 4 strains of control strains tested negative. The specificity of PCR products was confirmed by DNA sequencing. In addition, the minimum detection limit of DNA was 3.58 ng of SEB and the whole process was less than 20 h. A rapid, specific and sensitive detection method was established and the method can simultaneous detect Staphylococcal enterotoxins A, B in the same reaction condition.
Publication Date
10-28-2015
First Page
59
Last Page
61
DOI
10.13652/j.issn.1003-5788.2015.05.014
Recommended Citation
Jichao, LIU; Bairu, CHEN; Tiemin, JIANG; and Lijun, CHEN
(2015)
"Research on degenerate primers for detection of Staphylococcus aureus enterotoxin A, B gene,"
Food and Machinery: Vol. 31:
Iss.
5, Article 14.
DOI: 10.13652/j.issn.1003-5788.2015.05.014
Available at:
https://www.ifoodmm.cn/journal/vol31/iss5/14
References
[1] 何晖，刘华章，魏泉德. PCR技术检测金黄色葡萄球菌肠毒素A、B、C、E基因的研究[J].中国卫生检验杂志，2012，22(7)：1 585～1 587.
[2] Smyth Davida S， Hartigan Patrick J， Meaney William J, et al. Superantigen genes encoded by the egc cluster and SaPIbov are predominant among Staphylococcus aureus isolates from cows, goats, sheep, rabbits and poultry[J]. Journal of Medical Microbiology, 2005, 54(4): 401～411.
[3] 刘继超，姜铁民，姜阿赤. 多重PCR检测金黄色葡萄球菌六型肠毒素基因的研究[J]. 食品科技，2012，37(6)：304～307.
[4] Gigaud O. Staphylococcal enterotoxin genes of classical and new types detected by PCR in France[J]. Int. J. Food Microbiol., 2002, 77(1～2): 61～70.
[5] Lovseth A, Loncarevic S, Berdal K G. Modified multiplex PCR method for detection of pyrogenic exotoxin genes in staphylococcal isolates[J]. J. Clin Microbiol., 2004, 42(8): 3 869～3 872.
[6] Ikeda T, Tamate N, Yamaguchi K, et al. Quantitative analysis of Staphylococcus aureus in skimmed milk powder by real-time PCR[J]. J. Vet. Med. Sci., 2005, 67(10): 1 037～1 041.
[7] 顾琳，黄平，宋衍燕，等. PCR检测食品中金黄色葡萄球菌肠毒素基因[J]. 中国食物与营养，2015，21(2)：17～19.
[8] Mehrotra M, Wang G, Johnson W M. Multiplex PCR for detection of genes for Staphylococcus aureus enterotoxins, exfoliative toxins, toxic shock syndrome toxin 1, and methicllin resistance[J]. Journal of Clinical Microbiology, 2000, 38(3):1 032～1 035.
[9] 王少峡，陈丽媛，张竞秋，等. 利用生物信息学资源设计简并引物[J]. 天津师范大学学报(自然科学版)，2006，26(2)：26～29.
[10] Moon J S, Lee A R, Kang H M, et al. Antibiogram and coagulase diversity in staphylococcal enterotoxin-producing Staphylococcus aureus from bovine mastitis[J]. Journal of Dairy Science, 2007, 90(4):1 716～1 724.
[11] Simon A L, Jean M D, Johan M, et al. Identification of potyviruses using the polymerase chain reaction with degenerate primers[J]. Journal of General Virology, 1991(73): 1 531～1 541.