Folding primer multiplex pcr malaria molecular diagnosis kit and its detection method

Abstract

The invention discloses a folding primer multiple PCR malaria molecule diagnostic kit and a detection method thereof. The kit is composed of a magnesium-containing template buffer and a magnesium-free single combination reagent, and comprises a complete set of reagents for DNA preparation and a folding primer PCR reaction. The folding primer takes plasmodium Cyt b gene as a target gene, designing is carried out on plasmodium and specific site, sensitivity and singularity of four plasmodium detected by folding primer PCR can be obviously increased, and reagent and test operation can be simultaneously simplified. The kit of the present invention is employed for detection, a traditional reagent complex operation of nested-set / multiple PCR is simplified to a single reagent and a single tube for one step amplification reaction, four plasmodium individual or mixing infection can be detected and identified simultaneously, minimum of detected monocontamination can reach or approach 1 plasmodium / muml blood; and four protozoon can be simultaneously detected and accurately identified when the mixing infection is high, and low density difference is 100 times. The diagnostic kit and the detection method is a reliable means for definite diagnosis and discriminating of malaria in hospitals and laboratory in province and city.

Description

technical field [0001] The invention relates to a folding primer multiple PCR malaria molecular diagnosis kit and a detection method thereof, which belong to the field of medical examination. Background technique [0002] There are four types of Plasmodium that infect humans: Plasmodium falciparum (PF), Plasmodium vivax (PV), Plasmodium malariae (PM) and Plasmodium ovale (PO), which in turn cause Plasmodium falciparum, Plasmodium vivax, malaria and malaria ovale. Malaria, AIDS, and tuberculosis are listed as three major global public health problems. The first two types of malaria were prevalent in my country. Since 2010, the incidence of malaria in the country has dropped to less than 1 / 100,000 for three consecutive years. my country has entered the stage of malaria elimination. Four species of Plasmodium (including different strains, mixed infection and low-density parasites) have continuously entered my country. In 2012, overseas imports accounted for 91.5% of the report...

AI Technical Summary

Problems solved by technology

[0007] Purpose of the present invention: in order to overcome existing nested PCR and detect 4 kinds of malaria parasite diagnostic experiment simultaneously, need multi-tube two-step amplification, test time is long and reagent, operation is complicated, need middle opening tube to add sample again, easy to pollute and Mistakes; and the rapid PCR of single-tube one-step reaction can only detect and identify the defects of two kinds of Plasmodium PF and PV; provide a single-tube one-step amplification that can simultaneously detect and identify single and mixed infections of 4 kinds of Plasmodium, Sensitive, accurate, simple and fast folding primer multiplex PCR malaria molecular diagnostic kit

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