NASBA method for detecting tomato spotted wilt virus

Abstract

The invention provides an NASBA method for detecting a tomato spotted wilt virus (TSWV). The sequences of NASBA primers for detecting the TSWV are SEQ ID No: 1-2 respectively. According to the highly conserved region of gene N of the TSWV, two inner primers with specificity are designed. The conserved gene sequences are shared by different strains with TSWV to ensure the reliability in detecting different sources of TSWV at the level of strains. The NASBA method is suitable for rapid detection and confirmation of TSWV and can be widely used in disease monitoring in production and environment, as well as TSWV confirmation in the import and export trade.

Description

Technical field [0001] The invention belongs to the technical field of plant pathogen detection, and specifically relates to a NASBA method for detecting tomato spotted wilt virus. Background technique [0002] In recent years, Tomato spotted wilt virus (TSWV) has been listed as one of the ten most harmful plant viruses in the world due to its wide host range and huge economic losses. In the 1960s and 1980s, the virus was pandemic on tobacco and tomatoes in Europe, America and Africa, with an annual incidence of 20% to 50%, causing losses of up to several billion US dollars. In Hawaii, Brazil, Italy, and South Africa, the prevalence of TSWV in the 1980s and 1990s caused crops such as tomatoes and lettuce to be almost extinct. In recent years, the tomato spotted wilt virus genus virus, especially TSWV, has become an important factor that causes great economic losses to a variety of economic crops and ornamental plants worldwide. [0003] Many vegetable seeds such as peppers, onion...

AI Technical Summary

Problems solved by technology

However, traditional TSWV detection and confirmation methods generally use biological hosts and morphological tests to determine whether plants have plant viruses. These methods are time-consuming and cumbersome to operate, and cannot meet the needs of disease control.

[0004] At present, the quarantine and identification of the virus is mainly limited to traditional detection techniques such as electron microscope technology, serological technology and RT-PCR, such as ELISA method, molecular hybridization, fluorescent quantitative PCR method, ordinary PCR method, etc., but in these methods, serum The detection sensitivity of science and hybridization technology is low, the operation steps are cumbersome, and the cycle is long; electron microscope and fluorescent PCR technology rely on large-scale instruments, so many laboratories cannot meet the requirements in terms of instrument configuration and detection capabilities

In addition, since tomato spotted wilt virus is a single-stranded RNA virus, which is easy to degrade, the complexity and sensitivity of the above methods limit the detection and prediction of the disease

Although sensitive using PCR technology, current testing practice shows that false positive or false negative detection is frequent

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