PCR fast detecting reagent kit and detecting method for monkeypox virus

Abstract

The invention relates to a PCR quick detection kit of a Monkey pox virus and a detection method thereof, comprising a reagent A, a reagent B, a PCR reagent tube, a TaqDNA polymerase, a kit consisting of the positive control of the pUC57 plasmid of a specific F3L gene segment which comprises the Monkey pox virus, and double-distilled water negative control for sterilizing. The detection result of the Monkey pox virus can be known in 3 to 4 hours by adopting the simple, quick and sensitive PCR detection kit, designing a specific primer according to a section of preserved sequence specially owned by the Monkey pox virus and carrying out augmentation comparison; the sensitivity is higher; and the lower detection limit is lower to the DNA concentration of 0.3pg, thus ensuring the accuracy and the specificity of the detection. The invention is especially suitable for the entry-exit inspection and quarantine for carrying out quick detection with high sensitivity on the Monkey pox viruses in the feeding rodents such as laboratory mice from America and Africa and the like, as well as the quadrumanas of monkey, rabbit and the products thereof.

Description

PCR rapid detection kit and detection method for monkeypox virus technical field The invention relates to a PCR detection kit for monkeypox and a detection method thereof, belonging to the field of biology. It is mainly used for highly sensitive and rapid detection of monkeypox virus in domesticated rodents such as experimental mice from the United States and Africa, monkeys and other primates, rabbits and their products. Background technique As early as 1958, it was discovered that a pox virus could cause a disease similar to human smallpox in experimental rhesus monkeys. This disease was called monkeypox, and the virus was named monkeypox virus accordingly. Human monkeypox cases were first reported in Zaire in 1970. The main features were blisters and pustules similar to smallpox, but the infectivity and pathogenicity of monkeypox virus were weaker than that of smallpox virus. Since then, outbreaks have been reported in countries such as Liberia, Cameroon, Ivory Coast, ...

AI Technical Summary

Problems solved by technology

In recent years, people have also done a lot of research on the detection of monkeypox virus by PCR method, such as Ropp, Lbrahim, etc., who used the hemagglutinin gene sequence amplification and restricted enzyme analysis of the amplified fragment to detect monkeypox Viruses, but enzyme digestion is required in the experiment, the results are not intuitive and time-consuming

Zhou Weiming, Kulesh et al. used the fluorescent quantitative real-time PCR method to detect monkeypox virus. Although compared with the conventional PCR method, the fluorescent quantitative PCR method does not require electrophoresis analysis, and because the addition of specific fluorescent-labeled probes can detect Real-time detection of PCR amplification products, which saves time and specificity, reduces cross-contamination, and can accurately quantitatively analyze multiple samples, but it also has obvious disadvantages, such as the need for a fluorescent PCR instrument, which is harmful to the laboratory. Equipment requirements are relatively high, and the cost is also relatively high. In addition, the fluorescent quantitative PCR method has high requirements for the design of primers and probes. In summary, although the fluorescent quantitative PCR method has been developed rapidly, it is still not as good as the conventional method. The PCR method is universal and convenient, and the present invention is exactly a test kit and a detection method that can detect monkeypox virus rapidly and highly sensitively developed on the basis of the conventional PCR method

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