摘要
根据GenBank已公布的化脓隐秘杆菌溶血素((Pyolysin,PLO)基因设计6条引物,建立一种灵敏的环介导等温扩增方法(loop-mediated isothermal amplification,LAMP)用于化脓隐秘杆菌(Arcanobacterium pyogenes)的快速检测,并对检测方法的特异性和灵敏度进行验证。对化脓隐秘杆菌、鼠伤寒沙门氏菌、非O1霍乱弧菌、单增李斯特氏菌、表皮葡萄球菌、金黄色葡萄球菌、嗜水气单孢菌、副溶血性弧菌、蜡样芽孢杆菌等9株实验菌株进行的特异性试验表明,建立的LAMP方法仅对化脓隐秘杆菌的检测结果为阳性;该方法的检测灵敏度在DNA水平上可达112 fg。采用本研究建立的方法检测20份林麝临床病例样品,共鉴定出10株化脓隐秘杆菌,与API Coryne生化鉴定方法的符合率为100%。
According to Arcanobacterium pyogenes plo gene sequences published on GenBank, six specific loop-mediated isothermal amplification （LAMP） primers were designed to establish a method of LAMP for detection of Arcanobacterium pyogenes from forest musk deer and the specificity and sensitivity of this method were assayed. The specific test results showed that positive results obtained only from Arcanobacterium pyogenes while the amplified results of 8 non-Arcanobacterium pyogenes strains including Salmonella typhimurium, non-O1 Vibrio eholerae, Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus, Aeromonas hydrophila, Vibrio parahemolyticus and Bacillus cereus were negative. The detection sensitivity of the LAMP assay for Arcanobacterium pyogenes was 112 fg pure genomic DNA per 25 μL reaction. The test based on the method of this essay were applied on 20 clinical samples, 10 clinical samples were positive which were in accord with API coryne identification.
出处
《湖南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2014年第3期316-320,共5页
Journal of Hunan Agricultural University(Natural Sciences)
基金
陕西省科学院应用基础专项(2013K–12)
关键词
林麝
化脓隐秘杆菌
环介导等温扩增(LAMP)
forest musk deer
Arcanobacterium pyogenes
loop-mediated isothermal amplification （LAMP）
