Monocyte hyperplasia Listeria PCR detecting method adding amplified interior label

Abstract

The invention relates to a detection method of Listeria monocytogenes PCR with monocyte hyperplasia used for adding internal amplification control for food safety, pertaining to the biological technical field. The invention comprises the steps: a section of internal amplification control sequence is synthesized artificially, and corresponding special primers are designed; the concentration of MgCl2 and the anneal temperature are optimally selected; PCR testing and comparison are made to choose an adding concentration which has least effect to detection sensitivity and is capable of clearly indicating the concentration of the internal amplification control of false negative; whether the specific and the internal amplification control of primers can be normally enlarged in the testing of a Listeria monocytogenes sample with the monocyte hyperplasia can be determined; the stability of a PCR testing system is estimated through interference-free experiments; whether the false negative of the internal amplification control is indicated or not is judged by testing the artificial pollution sample; the accuracy of the testing result of the PCR testing system is estimated. The invention improves the accurate rate of the PCR testing and provides effective and reliable technical means to investigate and test food-borne pathogenic bacteria required during quarantine and law enforcement process.

Description

technical field [0001] The invention relates to a detection method in the field of biotechnology, in particular to a PCR detection method for Listeria monocytogenes with the addition of an internal standard for food safety. Background technique [0002] PCR, the abbreviation of polymerase chain reaction in English, refers to: polymerase chain reaction. Listeria monocytogenes (Listeria monocytogenes) is an important food-borne pathogenic bacteria. Listeria cellulogenes is mainly spread and infected human body through food (especially animal food). Compared with diseases caused by other food-borne pathogens such as salmonella, food-borne listeriosis is a very serious disease with a mortality rate as high as 20-30%. It is listed as a key detection by the WHO food safety work plan One of the four major foodborne pathogens (Salmonella, Escherichia coli 0157:H7, Vibrio parahaemolyticus, and Listeria monocytogenes). Data from my country's foodborne disease monitoring network sho...

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However, the fluorescence quantitative method has high requirements on the instruments and equipment, and additional fluorescent pr

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