Eightfold PCR amplification kit and amplification primer thereof for detecting pathogenic bacteria in food

Abstract

The invention discloses an eightfold PCR amplification kit and an amplification primer thereof for detecting pathogenic bacteria in food, wherein a PCR amplification reaction solution consists of the following components: 0.4umol / L-0.7umol / L of a primer, 50-100mmol / L of Tris-HCl which is 8.0-8.5 in pH value, 50-100mmol / L of potassium chloride, 25-50mmol / L of magnesium chloride, 2-10mmol / L of dithiothreitol, 10%-15% of DMSO, 2.5-5.0mmol / L of dNTP, 0.1-0.2ug / uL of LBSA, 1% of TritonX-100, 30-60mmol / L of ammonium sulfate and 5U / uL of Taq enzyme. The amplification kit and the amplification primer disclosed by the invention can be used for synchronously detecting seven pathogenic bacteria; and the amplification kit and the amplification primer are high in sensitivity and high in specificity, and are capable of improving detection speed and reducing detection cost.

Description

technical field [0001] The invention relates to the technical field of detection of food pathogenic bacteria, in particular to an eight-fold PCR amplification kit for detecting pathogenic bacteria in food and amplification primers thereof. Background technique [0002] Food safety is one of the most concerned issues in today's society. Foodborne diseases and food poisoning are caused by pathogenic bacteria contaminating food. Common pathogenic bacteria that cause foodborne diseases include: Salmonella, Vibrio parahaemolyticus, Staphylococcus aureus , Listeria monocytogenes, Escherichia coli O157, Escherichia coli H7, Vibrio cholerae and Vibrio vulnificus, etc. At present, the multiplex PCR method is often used in the molecular diagnosis of multiple pathogenic bacteria in food or food-borne diseases at the same time, because this method can detect 3 to 4 kinds of pathogenic bacteria at the same time, which not only improves the detection speed but also reduces the inspection ...

AI Technical Summary

Problems solved by technology

However, in the traditional multiplex PCR system, due to the existence of multiple pairs of primers, the interference between primers and the mismatch between primers and templates lead to problems such as decreased sensitivity and increased non-specific amplification reactions, which restrict the development of multiplex PCR technology.

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