Viral hemorrhagic sepsis virus visualized nucleic acid test strip detection primer set and detection method

Abstract

The invention discloses a VHSV visual nucleic acid strip detection primer group and detection method. The sequences of primers are VHSV281-F:5'-FITC-CAGGCGTTGTCCGTGCTTCT-3', 5' terminal marking FITC and VHSV281-R:5'-biotin-TCCCCGAGTTTCTTGGTGATGTA-3',5' terminal marking biotin. The detection method comprises the following steps of (1) RT-PCR amplification; (2) PCR product test strip detection test. The method has the advantages that the operation is convenient, the specificity and the sensitivity are high, the result is intuitive, stable, reliable and environment-friendly and the like and is suitable for VHSV rapid screening of the port quarantine laboratory, the identification working efficiency of the entry and exit port is improved, the requirement for fast customs clearance on import and export trades can be well met, a used RT-PCR product detection test strip is a general test strip and can be widely used for development of PCR diagnostic reagents, no instruments and equipment such as an electrophoresis apparatus and a transilluminator are used in the detection method, the application cost is low, and the method has a great promoting effect on popularization and application of the PCR detection method to base laboratories.

Description

Technical field: The invention relates to a primer set and a detection method for visual nucleic acid test strip detection of viral hemorrhagic sepsis virus (VHSV). Background technique: Viral hemorrhaic septicaemia (viralhaemorrhaic septicaemia, VHS) is a severe infectious disease caused by rhabdoviruses in dozens of fish species such as salmon, trout, pike and turbot. %~100%. The disease is mainly prevalent in the northern hemisphere, and it is a disease required by the World Organization for Animal Health (OIE) to be declared. my country lists it as a second-class disease. The VHSV genome is a single-stranded negative-strand RNA whose linear genome encodes 5 proteins, including: nucleocapsid protein (N protein), two structural proteins (M1 and M2 proteins), glycoprotein (G protein) and RNA polymerase (L protein). VHSV particles are about 180nm long and 70nm wide. Its genome is about 12k bases in length. At present, the commonly used methods for detecting viral hemor...

AI Technical Summary

Problems solved by technology

The result of cell culture isolation VHSV method is reliable, but the detection cycle is long, which is not suitable for rapid differential diagnosis; the traditional virus cell culture method and serological method are limited by their own characteristics, the detection is time-consuming and laborious, and it is difficult to meet the needs of rapid detection

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