Anisakis real-time fluorescent PCR detection kit and detection method

Abstract

The invention provides a real-time fluorescence PCR detection kit of anisakis. The kit comprises a specific PCR primer and a fluorescent probe, wherein the specific PCR primer comprises an anisakis ribosome 18s forward primer having a base sequence shown by SEQ. ID. No. 1 in a sequence table and an anisakis ribosome 18s reverse primer having a base sequence shown by SEQ. ID. No. 2 in the sequence table; the fluorescent probe has a base sequence shown by SEQ. ID. No.3 in the sequence table, the 5 ' terminal of the fluorescent probe is marked by a fluorescent reporter group, and the 3' terminal is marked by a fluorescence quenching group. The kit provided by the invention reflects a target gene amplification process in real time and directly reflects the copy number of the target gene through according to internal reference or external reference; the kit has the advantages of high sensitivity, strong specificity, short period, high throughput and the like, and meanwhile, the complicated operation of gel electrophoresis is omitted due to the optical system for detecting fluorescence signal, thereby both reducing the occurrence rate of false positive and potential harm on human and environment, and shortening the detection time.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, and in particular relates to a real-time fluorescent PCR detection kit and detection method of anisakis. Background technique [0002] With the rise of the diet fashion of eating raw fish, anisakiasis has become an important foodborne zoonotic parasitic disease. In order to understand the infection status of Anisakis in seafood, it is necessary to detect Anisakis on the seafood in the market. However, the traditional detection method mainly uses morphological identification technology, which is cumbersome, time-consuming and low detection rate. [0003] Compared with the shortcomings of traditional detection methods, the existing biomolecular detection methods are mostly carried out through the steps of pretreatment of the side sample, nucleic acid extraction, gel electrophoresis, etc., but the operation of gel electrophoresis during the implementation of this method is prone...

AI Technical Summary

Problems solved by technology

However, the traditional detection method mainly uses morphological identification technology, which is cumbersome, time-consuming, and low detection rate.

[0003] Compared with the shortcomings of traditional detection methods, the existing biomolecular detection methods are mostly carried out through the steps of pretreatment of the side sample, nucleic acid extraction, gel electrophoresis, etc., but the operation of gel electrophoresis during the implementation of this method is prone to occurrence False positive results, and the process is cumbersome

Images