Yellow fever virus detection kit based on RPA technology and applications of yellow fever virus detection kit

Abstract

The invention discloses a yellow fever virus detection kit based on the RPA technology and applications of the yellow fever virus detection kit. The experiment proves that an RPA primer and a probe of the yellow fever virus can effectively amplify a target gene, the specificity achieves 100%, the sensitivity is 1x10<2> copies / reaction, the virus titer capable of being detected in a clinical simulation sample is 10<2>TCID50 / mL, and the sensitivity is equivalent to that of the fluorescent quantitative PCR; no cross reaction with the congeneric dengue virus, encephalitis B virus and tick-borne encephalitis virus nucleic acid exists. For the RPA isothermal amplification system provided by the invention, the reaction is rapid, the temperature range is wide, under the temperature of 37-42 DEG C, the effective amplification of the target gene can be realized, the yellow fever virus detection kit can be used for the on-site rapid detection for nucleic acid infected with the yellow fever virus, and meanwhile, an available rapid detection method is provided for the on-site screening of pathogenes. The yellow fever virus detection kit has great significance for preventing the infection and propagation of the yellow fever virus in China, and for the inspection and quarantine of epidemic areas and entry and exit ports.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a yellow fever virus detection kit based on RPA technology and an application thereof. Background technique [0002] Yellow fever virus (Yellow fever virus) belongs to the Flavivirus genus of the Flaviviridae family, can cause severe hemorrhagic fever symptoms, is highly contagious, and has a case fatality rate of 23%. ～ 90%. The main manifestations are acute fever accompanied by severe bleeding. Yellow fever is mainly prevalent in Africa. China is not an endemic area for yellow fever virus. However, with the rapid development of global trade and tourism, human exchanges between different countries and regions have become increasingly frequent. Many infectious diseases have broken the borders of regions and countries. The scope of transmission is becoming wider and wider, and our country has also faced the threat of imported infectious diseases many times. In March 201...

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Problems solved by technology

[0003] At present, human detection of yellow fever virus mainly relies on real-time fluorescent quantitative PCR technology, which requires expensive temperature control equipment, specific laboratory space and professional technicians to operate, and the detection time is long, so it is not suitable for on-site rapid detection of sudden infectious diseases Therefore, establishing a quick and easy nucleic acid detection technology is of great significance and value for the timely discovery and identification of yellow fever virus

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