On-site rapid high-sensitivity detection kit of prawn infectivity muscle necrosis virus and detection method thereof

Abstract

The invention relates to an on-site rapid high-sensitivity detection kit of prawn infectivity muscle necrosis virus and a detection method thereof. The detection kit comprises a sampling tube, a rinsing tube filled with distilled water, a nucleate degeneration tube filled with a TE buffer solution, an amplification detection tube filled with amplification reaction liquid and nucleic acid dye, a negative control tube, a positive control tube, an FTA membrane, rapid drying liquid, and the like. Compared with the RT-PCR detection method, the detection method in the invention has the advantages of higher specificity, sensitivity and convenience, low cost, convenient use, more accurate and rapid detection and extremely sensitivity, is safe to both human and the environment and can substitute the traditional relevant detection method. The invention can be used in both the indoor laboratory and outdoor production field, has great significance for strengthening the epidemiological monitoring of prawn infectivity muscle necrosis virus, prawn cultivation waterbody monitoring and disease prevention and control and has great promotion and application value.

Description

technical field [0001] The invention relates to a marine biological pathogen detection technology, in particular to an on-site rapid and high-sensitivity detection kit for shrimp infectious myonecrosis virus (IMNV) and a detection method thereof. Background technique [0002] Infectious myonecrosis is a new viral infectious disease that endangers the production of prawns found in Brazil in 2002. In 2004, Dr. Donald V. Lightner of the University of Arizona found that the disease was caused by a new virus and Name it Infectious myonecrosis virus (IMNV for short). The novel virus is a double-stranded RNA virus belonging to the family Holoviridae. The susceptible prawn species of prawn infectious myonecrosis virus is Penaeus vannamei, mainly juvenile shrimp infected 60-80 days old. The virus can cause muscle tissue necrosis in the whole body of susceptible prawns. Generally, pathological death occurs slowly and the mortality rate is not high, but there are deaths throughout th...

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Problems solved by technology

The pathological section method cannot directly detect the virus, but can only detect the histopathological signs of the disease, and it is also limited to the laboratory; although the electron microscope technology can directly observe the existence of virus particles, its operation is complicated and costly. The time is long and the accuracy is low; although the detection of IMNV by the antibody detection method is faster than the pathological section method, its sensitivity is low, and it is difficult to detect by the antibody method before IMNV has caused infection or the very early stage of infection; the RT of IMNV -PCR detection method, although it overcomes the shortcomings of the previous methods, it can achieve relatively fast and accurate detection of IMNV under laboratory conditions, but because conventional RT-PCR detection requires expensive PCR machines, gel electrophoresis and imaging system, making it difficult for RT-PCR detection method to be used for rapid detection on the spot, which greatly limits the application of RT-PCR detection method in production