Newcastle disease virus LAMP detection reagent case and detection method thereof

Abstract

The invention relates to a reagent box and a detecting method for quickly detecting newcastle disease virus by using loop-mediated isothermal amplification (LAMP) technique, which comprises 10 x isothermal reaction buffer solution, AMV 8U / Mul, Rnasin 40U / Mul, Bst DN polymerase 8U / Mul, 10mM dNTP, 100mM magnesium sulphate, 20 Mu M inner primer 1, 20Mu M inner primer 2, 10Mu M outer primer 1, 10Mu outer primer 2, the reaction solution A of 5M betaine, and the reaction solution B of the betaine: 1000 x fluorescent dye SYBR Green I. By extracting the RNA of tissue sample as well as the RNA in the poultry throat and cloaca cotton swab sample, conducting the loop-mediated isothermal amplification of newcastle disease virus, and detecting the coloration of the amplified products, the newcastle disease virus can be detected. By eliminating the defects in prior art, such as time-consuming quality, large workload, cross contamination and complex operations, the reagent box and the detecting method for quickly detecting newcastle disease virus has the advantages of strong specificity, high sensitiveness, good speed, low cost, simpler operating methods, and perfect applicability to quick detection at site.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a kit for rapidly detecting Newcastle disease virus using a loop-mediated isothermal amplification (LAMP) technology, and a method for rapidly detecting Newcastle disease virus using the kit. Background technique [0002] Newcastle disease (ND) is an acute and highly contagious poultry disease caused by Newcastle disease virus (NDV), which is a serious hazard to the poultry industry. The World Organization for Animal Health (OIE) lists it as a statutory notifiable infectious disease, and my country also defines it as a first-class animal infectious disease. [0003] With the development of highly intensive poultry farming, the control of ND has attracted much attention. The on-site diagnosis of ND mainly relies on the comprehensive judgment of history, clinical symptoms, pathological changes and epidemiological investigation, which has brought overload work to the majority of veterina...

AI Technical Summary

Problems solved by technology

These methods take a long time to report results and require a lot of work

However, the commonly used polymerase chain reaction (PCR) technology requires special instruments, and has the disadvantages of easy cross-contamination and cumbersome operation.

Although fluorescent real-time quantitative polymerase chain reaction (real time PCR) technology has solved the problem of cross-contamination and simplified the operation process, it needs more complex quantitative measurement instruments, so it is not suitable for rapid on-site detection. Higher, it also increases the difficulty of promotion and application