A rt-lamp visualization kit for detecting goose Newcastle disease virus and its application

Abstract

The invention discloses an RT-LAMP visualkit for detecting goose newcastle disease virus (NDV) and application thereof. The visual kit comprises a primer group and color materials, wherein the primer group is a sequence as shown by SEQ ID No: 1 to 4, and the color materials are calcein and manganese chloride. An application method of the kit comprises the following steps of firstly preparing an RT-LAMP reaction system, which comprises AMV reverse transcriptase, 10-time reaction buffer, strand displacement DNA polymerase, dNTP mixture, glycine betaine, the calcein, the manganese chloride, MgSO4, an FIP primer, a BIP primer, an F3 primer, a B3 primer and a sample RNA to be tested; then reacting the reaction system for 50min at the constant temperature of 61 to 65 DEG C, and finally inactivating for 4min at the temperature of 80 DEG C; interpreting a result through natural illumination or ultraviolet light or interpreting through the natural illumination and the ultraviolet light; if a reaction product is green under the natural light, indicating that the sample to be tested contains the NDV; if the reaction product shows obvious green fluorescence under the ultraviolet light, indicating that the sample to be tested contains the goose NDV.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an RT-LAMP visualization kit for detecting goose Newcastle disease virus and its application. Background technique [0002] Newcastle disease (ND) is an acute and severe infectious disease caused by Newcastle disease virus (NDV), characterized by respiratory and digestive tract lesions. sick. So far, there have been four ND pandemics worldwide, causing huge losses to the poultry industry. Waterfowl, as the natural reservoir of NDV, were once considered to have strong resistance to the virus, even if they were infected with a strong strain, they did not show any clinical symptoms. In 1997, since a highly pathogenic strain to geese was isolated in Jiangsu and Guangdong, it has continued to break out among geese flocks all over the country, causing a large number of deaths. At present, goose Newcastle disease (GND) has caused huge economic losses and seriously endangered the healthy ...

AI Technical Summary

Problems solved by technology

However, the use of a turbidimeter not only increases the cost, but is also inconvenient to carry; some scholars use the method of adding SYBR Green I dye to the reacted system to detect the amplification product, but this observation must be opened for processing. RT-LAMP The high efficiency and high sensitivity of amplification make the product contain a large number of target fragments, and adding dyes after opening the cap is extremely easy to pollute the environment

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