Shrimp health system visual rapid detection kit based on LAMP technology

Abstract

The invention discloses a shrimp health system visual rapid detection kit based on an LAMP technology, and belongs to the technical field of aquatic pathogen detection reagent development. The kit comprises an internal reference quality control primer group and a to-be-detected pathogen primer group. The kit is designed for common shrimp pathogenic microorganisms of IHHNV virus, white spot syndrome virus, enterocytozoon hepatopenaei, shrimp hemocyte iridescent virus, shrimp acute hepatopancreatic necrosis disease-vibrio parahaemolyticus and the like; the kit can be used for simultaneously detecting various pathogens; by adding the internal reference quality control primer group, the influence of failure of a reaction reagent and nucleic acid extraction quality of an amplification templateon a detection result can be eliminated; the accuracy of the detection result is ensured; and the kit is suitable for screening and purchasing young shrimps and parent shrimps by grass-roots farmers and customs rapid quarantine customs clearance.

Description

technical field [0001] The invention belongs to the technical field of aquatic pathogen detection reagent development, and in particular relates to a visual rapid detection kit for shrimp health system based on LAMP technology. Background technique [0002] China is the largest shrimp producer in the world; China's shrimp production accounts for about 40% of the world's total shrimp production; and the amount of shrimp farming is increasing year by year. However, shrimp farming diseases are also constantly occurring. At present, the most serious pathogens in shrimp aquaculture are: white spot syndrome virus (WSSV) of shrimp; Infectious hypodermal and hematopoietic necrosis virus (IHHNV); Shrimp hemocyte iridescent virus, SHIV); acute hepatopancreatic necrosis disease (AHPND); shrimp Enterocytozoon hepatopenaei (EHP). [0003] In addition, my country's broodstock is heavily dependent on imports; my country's shrimp seed industry introduces about 20% of the first-generation ...

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Problems solved by technology

There are few studies on the simultaneous detection of multiple pathogens; moreover, the amplification process of most existing LAMP detections is carried out in a reactor similar to a PCR instrument, the detection cost is high for users, and the detection results are not intuitive enough to be displayed visually. The measurement method is complicated

For example, Chinese patent application 201910090452.6 discloses a high-throughput quantitative detection kit for aquaculture animal viruses, which includes a LAMP detection primer set for 14 kinds of aquaculture animal viruses. The detection of 14 kinds of aquaculture animal viruses within 3 hours has the advantages of high throughput and rapid detection; however, this method does not get rid of the dependence on expensive instruments, and is not suitable for customs quarantine and the promotion and use of grassroots farmers

[0005] In addition, because the LAMP reaction is easily affected by factors such as detection reagents (such as reaction enzyme inactivation), nucleic acid extraction quality of the amplified template (such as the extracted nucleic acid is severely degraded or the nucleic acid is not extracted due to operational errors), therefore, setting The internal reference quality control of LAMP reaction detection is particularly important; at present, the reports on the internal reference quality control are mostly found in human disease detection, but there are few reports on the internal reference quality control of the detection of aquatic pathogens

Moreover, most of the existing internal reference quality control primer sets are only aimed at the same organism, and there are few reports on internal reference quality control primer sets suitable for multiple organisms.

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