Method and kit for quickly detecting interior label of salmonella by adopting reverse transcription fluorescent quantitation

Abstract

The invention discloses a quick detection method by using an interior label and taking a sigD / sigE gene of salmonella as a target through reverse transcription fluorescent quantitation PCR (Polymerase Chain Reaction). In the quick detection method, an RNA (Ribonucleic Acid) interior label obtained by PCR amplification for three times is composed. In addition, the invention further provides a corresponding kit and application. The method disclosed by the invention can be widely applied to the fields such as research of laboratories, food safety and health and hygiene. The phenomenon of false negative missed detection can be avoided, so that the PCR amplification is effectively monitored; in addition, the detection specificity of salmonella is strong, so that dead bacteria and live bacteria can be effectively distinguished; the quickness and high detection sensitivity are obtained; and the method can be applied to high-throughput screening of the salmonella in the food.

Description

technical field [0001] The invention belongs to a nucleic acid detection method in the field of food safety. Specifically, the present invention relates to a method, a kit, and an application for rapid detection of internal standard reverse transcription fluorescence quantitative detection of Salmonella sigD / sigE gene band. Background technique [0002] Salmonella (Salmonella spp.) is one of the important food-borne pathogens, which is mainly transmitted and infected through meat, milk, eggs and other foods, causing typhoid fever, sepsis, acute gastroenteritis and other diseases. Among the cases of bacterial food poisoning in various countries in the world, food poisoning cases caused by Salmonella often rank first or second. Establish an effective quantitative detection method for food-borne Salmonella, in order to improve the detection speed and accuracy of the bacteria, so that the contaminated food can be processed in time, and it is widely used in public health, food h...

AI Technical Summary

Problems solved by technology

Traditional biochemical detection methods can distinguish dead bacteria from live bacteria, but it takes a long time; and using real-time fluorescent quantitative PCR (real-time PCR, referred to as RT-PCR) method, there is no effective method to eliminate false positives caused by inhibitors. Negative problems, so it is necessary to develop products that effectively monitor false negatives to avoid missed detections

Images