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CN108103212A 一种环介导等温扩增快速检测沙门菌的方法 无效 - 驳回

一种环介导等温扩增快速检测沙门菌的方法
摘要：
一种环介导等温扩增快速检测沙门菌的方法，步骤如下：提取待测菌的基因组DNA：以提取的基因组DNA为模板进行LAMP扩增：扩增反应体系中硫酸镁的终浓度为6.5~8 mM/L，dNTPs的终浓度为4.2~5mM/L，F3和B3引物的终浓度为0.13μM/L~0.26μM/L，FIP和BIP引物的终浓度为1.2~2μM/L，Bst DNA聚合酶的终浓度为0.3~1.8 U/μL，Betaine的终浓度为1~2M/L，扩增温度为60~62℃1~1.5h,85~87℃1~3min。本发明引物特异性强，根据是否扩增即可判断检测样品是否存在沙门氏菌，同时具有灵敏度高，可快速完成检测，成本较低。
摘要（英）：

The invention discloses a method for quickly detecting salmonella through loop-mediated isothermal amplification. The method comprises the following steps: extracting the genome DNA (deoxyribonucleicacid) of bacteria to be detected: taking the extracted genome DNA as a template to carry out LAMP (Loop-mediated isothermal amplification), wherein the final concentration of magnesium sulfate in an amplification reaction system is 6.5-8mM/L, the final concentration of dNTPs (High quality deoxyribonucleotide triphosphates) is 4.2-5mM/L, the final concentration of F3 and B3 primers is 0.13-0.26mu m/L, the final concentration of FIP (Forward Inner Primer) and BIP (Backward Inner Primer) primers is 1.2-2 mu M/L, the final concentration of Bst DNA polymerase is 0.3-1.8U/mu L, the final concentration of Betaine is 1-2M/L, and amplification temperature is 60-62 DEG C for 1-1.5h and 85-87 DEG C for 1-3min. The method has the advantages of high primer specificity, high sensitivity and lower cost,whether a detected sample contains salmonella or not can be judged according to whether amplification is carried out or not, and detection can be quickly finished.

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