溶血性链球菌LAMP检测方法的建立

目的：研究溶血性链球菌的环介导等温扩增(loop-mediated isothermal amplification，LAMP)检测方法，实现对溶血性链球菌的快速检测。方法：针对溶血性链球菌的scpA 基因设计4 条特异性LAMP 内外引物进行LAMP扩增；优化扩增反应条件；采用包括溶血性链球菌在内的6 种不同菌株进行LAMP 的特异性检测，并酶切鉴定；对溶血性链球菌菌液以无菌水进行10 倍系列梯度稀释后，进行LAMP 扩增检测其灵敏度；将菌掺入牛奶样本并进行LAMP 检测。结果：本法可快速灵敏地检测出溶血性链球菌，反应特异性高，检出限为16.7CFU/mL，而牛奶样本的检出限则达10CFU/mL。结论：LAMP 法可用于食品溶血性链球菌的快速检测。

Establishment of Loop-mediated Isothermal Amplification for the Detection of Streptococcus hemolyticus

Objective: To develop a loop-mediated isothermal amplification (LAMP) method for the rapid detection of Streptococcus hemolyticus. Methods: According to the scpA gene of Streptococcus hemolyticus, 4 specific primers were designed for LAMP amplification. Six different Streptococcus hemolyticus strains were validated by LAMP and identified by enzymatic digestion. The sensitivity of LAMP method was evaluated by serial gradient dilution of Streptococcus hemolyticus solution. Results: LAMP could be used for the rapid and sensitive detection of Streptococcus hemolyticus with a high sensitivity and a detection limit of 16.7 CFU/mL. Moreover, the detection limit was 10 CFU/mL in milk samples. Conclusion: LAMP method is a simple, rapid and feasible method for the detection of Streptococcus hemolyticus.