Novel and practical litopenaeus vannamei RNA (ribonucleic acid) virus fast detection kit and detection method

Abstract

The invention discloses a novel and practical litopenaeus vannamei RNA (ribonucleic acid) virus fast detection kit and a detection method. The detection kit comprises a PBS (poly butylenes succinate) pretreatment solution, an RNA fast extraction solution and a detection solution. The kit and the detection method have the advantages that the sensitivity is high; the specificity is high; the detection time is short; the accuracy is high; the operation is simple; the requirements on instruments and equipment are low; the pollution cannot be caused. The problems that the conventional detection method has high requirements on the equipment and the detection environment, the operation is complicated, the requirement on the specialty of technicians is high, and the like are solved. The wide application prospects are realized.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a novel and practical rapid detection kit and detection method of Litopenaeus vannamei RNA virus. Background technique: [0002] Litopenaeus vannamei (Litopenaeus vannamei, also known as Penaeus vannamei) occupies an absolute dominant position in my country's prawn farming. In 2014, the aquaculture output reached 1.4 million tons, accounting for more than 85% of the country's total prawn aquaculture output. It occupies a relatively large share in China's fishery economy. Large specific gravity. However, disease problems cause huge losses to the shrimp industry every year, seriously restricting the healthy development of the industry. Litopenaeus vannamei RNA virus is one of the more harmful species, such as: Taura virus (TauraSyndromeVirus, TSV) can cause 60-90% of catastrophic mortality of Litopenaeus vannamei juveniles, stealing dead disease Nomura virus (Covertmortali...

AI Technical Summary

Problems solved by technology

However, there are certain deficiencies in the existing technical methods: for example, the detection sensitivity of the cell culture method is not high, and the detection time is long; the PCR detection method requires specific instruments and equipment, and has high requirements for the detection environment; Operation, higher professional requirements for technicians

However, the existing RNA virus LAMP detection kits all need to undergo conventional RNA extraction and purification processes. The RNA extraction method needs to undergo Trizol extraction, chloroform layering, isopropanol extraction and centrifugation, 100% ethanol, 75 % ethanol, DEPC water to dissolve RNA and other steps, these operations have high requirements for experimental equipment and technical personnel, which is not conducive to the popularization and application of LAMP reaction

In addition to adding various LAMP reaction primers, enzymes, reaction buffers and other steps, the reaction test tube needs to be opened many times to add reagents, resulting in the contamination of LAMP reaction reagents and false positive results.

Due to the above limiting factors, the application performance of the current LAMP detection kit is very limited

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