Application and method of imp gene in preparation of kit for detecting Riemerella anatipestifer serotype 1

Abstract

The present invention relates to the application and method of an Imp gene in the preparation of a kit for detecting Riemeria anatipestifer serotype 1. The specific steps are as follows: according to the Imp gene sequence SEQ ID NO.1 in the Riemeria anatipestifer genome DNA sequence Design the amplification primer pair sequence SEQ ID NO.2 and SEQ ID NO.3, add FAM fluorescent markers at the 5' and 3' ends at the same time, extract sample DNA, and amplify by PCR; DNA recovery kit for liquid recovery PCR amplification Store the product in the dark at ‑20°C for later use; use PNA synthesized with the sequence SEQ ID NO.4 as a probe for reverse hybridization, observe under a fluorescent microscope, if green fluorescent spots appear, it is the serum of Riemeria anatipestifer Type 1; if no green fluorescent spots appear, it is other strains. The detection method of the invention is used to detect the Riemerella anatipestifer serotype 1, and the detection time is short, the cost is low, the detection result is specific, and the result determination is simple.

Description

technical field [0001] The invention belongs to the field of livestock and poultry detection, and relates to a kit for detecting Riemerella anatipestifer serotype 1 by reverse hybridization using a PNA probe, and also relates to a detection method using the kit. Background technique [0002] Riemerella anatipestifer (RA) is the causative bacterium of infectious serositis in ducks, which is one of the most important diseases that endanger the duck industry. The disease is an acute or chronic sepsis process, characterized by fibrinous pericarditis, perihepatitis, air sac inflammation, meningitis, and some caseous salpingitis and arthritis. The morbidity rate is 10%-90%, and the case fatality rate is as high as 80%, which can occur throughout the year. The disease is prevalent in almost all duck raising areas in the world, causing huge economic losses to the duck industry. The traditional identification of RA usually detects phenotypic indicators such as its cultural characte...

AI Technical Summary

Problems solved by technology

The disease is prevalent in almost all duck raising areas in the world, causing huge economic losses to the duck industry

The traditional identification of RA usually detects phenotypic indicators such as its cultural characteristics, morphological staining, physiological and biochemical characteristics, and certain chemical compositions of the bacteria. so confused

In addition, the traditional identification method is complicated to operate, time-consuming and laborious, which is not conducive to timely diagnosis of the cause, inspection of the pathogen and control of the spread of the disease

Aiming at these characteristics of Riemerella anatipestifer, many researchers have established some detection techniques, such as fluorescent antibody technique, ELLSA, immunohistochemistry, etc., which have been reported successively, but in practice, they all have different degrees of limitations, such as the need to target different serum type of antibody as a detection reagent

Images