RPA primer and probe for detecting Listeria monocytogenes and detection method

Abstract

The invention provides a RPA primer and a probe for detecting Listeria monocytogenes and a detection method. The RPA primer and the probe are designed based on a Listeria monocytogenes actA gene, andprimer sequences are shown in SEQ ID No.1, 2 and 3. The probe length is 45-52bp, a position where is 30bp apart from 5' terminal in the middle of the probe is modified by dSpacer, thymidine at doublesides of the dSpacer molecule are respectively replaced by a fluorescence group FAM and a quenching group BHQ1, a 3' terminal of the probe is modified by a blocking group C3Spacer, and the sequence isshown in SEQ ID No.4. Compared with the prior art, detection by the detection method can be completed within 15 minutes, in order to rapidly detect whether Listeria monocytogenes exists in a sample;the established RPA method has good specificity and high sensitivity, special apparatuses are not needed, and the method has important application values in the fields of disease monitoring, clinic diagnosis, foodstuff safety monitoring, and the like.

Description

technical field [0001] The invention belongs to the field of molecular biology detection, and in particular relates to a primer, a probe and a detection method for detecting Listeria monocytogenes. Background technique [0002] Listeria monocytogenes is one of the most important foodborne pathogens and a zoonotic pathogen. After infection, the main manifestations are sepsis, meningitis and mononucleosis. It exists widely in nature. Listeria monocytogenes in food is dangerous to human safety. The bacteria can still grow and reproduce in an environment of 4°C. It is one of the main pathogenic bacteria that threaten human health in refrigerated food. Therefore, It is of great significance to develop a rapid detection method for Listeria monocytogenes. [0003] The current detection methods for Listeria monocytogenes are traditional culture methods, immunological methods and PCR methods. Although the traditional culture method has low requirements on experimental equipment an...

AI Technical Summary

Problems solved by technology

Although the traditional culture method has low requirements for experimental equipment and strong operability, the detection cycle is long and the detection efficiency is low, which cannot meet the requirements of rapid detection of samples in the current detection work.

However, immunological detection methods have non-specific reactions, although they have high sensitivity, they lack specificity.

The disadvantage of the PCR detection method is that the amplification reaction is affected by many factors, requires relatively expensive instruments, is prone to non-specific amplification, and the detection time is relatively long

At present, RPA technology has been applied to the rapid detection of viruses, bacteria, mycoplasma, parasites, etc., but it has not been applied to the detection of Listeria monocytogenes.

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