Quickly-nested PCR (polymerase chain reaction) malaria molecular diagnosis reagent kit and detection method

Abstract

The invention discloses a quickly-nested PCR (polymerase chain reaction) malaria molecular diagnosis reagent kit and a detection method. The quickly-nested PCR malaria molecular diagnosis reagent kit comprises novel primers with a self-closing function, cytochrome oxidase genes (cox1) in a plasmodium mitochondrial genome serve as target genes, and four bidirectional semi-nested primers are designed in loxp sites of plasmodium groups and species to form a pair of group-specific primers and two species-specific primers. A traditional nested / multiplex PCR complicated in reagents and burdensome in operation is simplified to single-reagent and single-tube one-step reaction, separate infection or mixed infection of malignant malaria and plasmodium vivax and separate infection of quartan malaria and ovale malaria can be detected and authenticated simultaneously, sensitivity and specificity of the detection method are superior to a microscopic examination method, and the level of detection of one worm / per microliter blood can be achieved. The detection method serves as a reliable means for diagnosis and differential diagnosis of malaria of hospitals at all levels, and the quickly-nested PCR malaria molecular diagnosis reagent kit is a simple, economic, sensitive and accurate malaria diagnosis tool for control and monitoring of malaria, malaria medicine tests, malaria vaccine research and blood donor screening.

Description

technical field [0001] The invention belongs to the field of medical testing, and relates to a rapid nested PCR malaria molecular diagnosis kit and a detection method thereof. Background technique [0002] Malaria is one of the three most serious infectious diseases (AIDS, tuberculosis and malaria) in the world. The World Health Organization (WHO) has listed malaria control as the action goal of this century. There are four kinds of Plasmodium that infect humans: Plasmodium falciparum, P.vivax, P.malariae and P.ovale. Due to the biological and immunological characteristics of the four Plasmodium species, their pathogenicity, therapeutic effect of antimalarial drugs, susceptibility to the vector Anopheles mosquitoes and their distribution in the regions of global prevalence vary. Every year, hundreds of millions of malaria patients are infected worldwide, and more than one million of them die from falciparum malaria. Timely and accurate diagnosis and differential diagnosis ...

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention: to overcome the traditional nested PCR malaria diagnostic test that requires multiple tubes and two-step amplification, the test time is long, and the reagent operation is complicated, the tube needs to be opened in the middle and the sample is easily polluted, and non-specific amplification such as dimer is seriously affected Defects such as result judgment; standardize reagents and operations, uniformly prepare combined reagents, and complete most of the reagent operations in the kit prefabrication stage to ensure rapid test, repeatability and reduce detection costs, providing a rapid nested PCR malaria Molecular diagnostic kit and detection method thereof

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