应用多重PCR技术检测120株蜡样芽胞杆菌毒力基因  

Detection of 120 strains of Bacillus cereus virulence genes by multiplex PCR

作　　者：刘秀峰[1] 潘洁茹[1] 林萍[1] LIU Xiu -feng, PAN Jie -ru, LIN Ping(Fuzhou Center for Disease Control and Prevention, Fuzhou, Fujian 350004, China)

机构地区：[1]福州市疾病预防控制中心福建医科大学教学基地,福建福州350004

出　　处：《现代预防医学》2018年第21期3972-3976,共5页Modern Preventive Medicine

基　　金：福建省福州市科技计划项目(No.2014-s-145)

摘　　要：目的应用多重PCR技术对蜡样芽孢杆菌毒力基因进行快速检测,为研究蜡样芽胞杆菌致病性提供有力依据。方法根据蜡样芽胞杆菌(B. cereus)的cer、hbl A、hbl C、nhe A、nhe B、cyt K和bce T基因设计并合成引物,优化反应条件,应用多重PCR技术检测120株B. cereus毒力基因序列。结果多重PCR检测结果非常理想,不同基因扩增条带清晰可辨,蜡样芽胞杆菌呕吐毒素基因cer携带率为96. 66%(116/120),细胞毒素cyt K基因携带率为42. 37%(50/118),肠毒素bce T基因携带率为38. 33%(46/120),非溶血性基因nhe A携带率为78. 33%(94/120),非溶血性基因nhe B携带率为83. 05%(98/118),溶血性基因hbl A携带率为55. 0%(66/120),溶血性hbl C基因携带率为27. 96%(33/118)。结论多重PCR结果很好,7对引物所扩增出的DNA条带区分明显,电泳条带清晰,特异性强,敏感度高。本实验对120株蜡样芽胞杆菌7种毒力基因进行检测,为研究蜡样芽胞杆菌致病性提供有力依据。Objective To use multiple PCR assay for the detection of enterotoxic Bacillus cereus group strains, and to provide strong evidence for the pathogenicity study of Bacillus cereus. Methods 7 pairs of primers were designed and compounded according to the B. cereus＇ s gene of car, hblA, hblC, nheA, nheB, eytK and beeT. The reaction conditions were optimized, and 120 strains of Bacillus cereus virulence gene sequence were detected by multiplex PCR. Results The bands of 7 pairs of primers were clear and distinguishable. The carrier rate of vomiting toxin gene cer in Bacillus cereus was 96.66% （ 116/120）, the carrying rate of cytK gene was 42.37 % （50/118 ）, and the carrying rate of enterotoxin bceT gene was 38.33% （46/120）. The carrying rates of hemolysin hblA, hblC and the nonhemolytic enterotoxin nheA, nheB were 55.0% （66/120）, 27.96% （33/118）, 78.33% （94/120） and 83.05% （98/118）, respectively. Conclusion The results of this study show that mPCR increases the detection efficiency. The DNA bands amplified by the seven pairs of primers are distinct, the electrophoresis bands are clear, the specificity is strong, and the sensitivity is high. In this experiment, the detection of seven virulence genes of 120 strains of Bacillus cereus provides a powerful basis for studying the pathogenicity of Bacillus cereus.

关 键 词：蜡样芽孢杆菌 多重PCR技术 毒力基因 腹泻 携带率 

分 类 号：R113[医药卫生—公共卫生与预防医学]