Plasmodium detection kit

Abstract

The invention provides a plasmodium detection kit. The plasmodium detection kit comprises PCR (Polymerase Chain Reaction) reaction liquid for fluorescence quantitative PCR detection, wherein particularly, the PCR reaction liquid comprises a forward primer sequence and a reverse primer sequence for target polynucleotide amplification; the forward primer is 5'-CGACTAGGTGTTGGATGAAAGTGT-3'; a reverse primer is 5'-CCAGAACCCAAAGACTTTGATTTC-3'. In a specific implementation manner, the PCR reaction liquid comprises a probe sequence for target polynucleotide detection, wherein the probe sequence is 5'-TTCGAGGTGACTTTTAGATTGCTTCCTTC-3'. The plasmodium detection kit provided by the invention is simple and convenient to operate, high in sensitivity and good in specificity, repeatability and stability, and is capable of preventing PCR product pollution; fluorescence quantitative PCR detection of plasmodium is carried out by utilizing the plasmodium detection kit; whether the plasmodium exists in a blood sample or not can be determined; and reliable experimental evidences can be provided for determining whether sensitive and early diagnosis is infected by plasmodium or not.

Description

technical field [0001] The invention belongs to the technical field of detection kits, in particular to a fluorescent quantitative PCR detection kit for malaria parasites. Background technique [0002] Malaria is one of the most serious tropical diseases caused by Plasmodium in the world, infecting about 400 million people every year, most of which occur in countries such as sub-Saharan Africa (Plasmodium falciparum) and Southeast Asia (Plasmodium vivax). Malaria costs Africa as much as $12 billion a year in economic losses. In the treatment of malaria, the early detection of the cause is particularly important. Now the mature commercial malaria parasite detection reagents at home and abroad are mostly immunological platform products. [0003] The detection of Plasmodium mainly involves two aspects, nucleic acid extraction and nucleic acid amplification detection. [0004] Among domestic mature products, there are currently three main methods for extracting nucleic acid in...

AI Technical Summary

Problems solved by technology

The advantage is that it is easy to operate; the disadvantage is that it cannot effectively remove the factors that inhibit PCR in peripheral blood, and weak positives often have no amplification

2) Concentrated boiling method: first concentrate the peripheral blood, then add the lysate, boil, high-speed centrifuge, and use the supernatant as a template. This method is currently a common clinical method in China. Its advantage is that it can partially remove the Inhibitory factors that cannot be removed; the disadvantage is that different manufacturers have different concentration effects, some can see precipitation, and some cannot

The Plasmodium extraction and detection methods provided by these existing kits have many deficiencies: 1) the nucleic acid extraction process is more complicated, the sample processing takes a long time, and when processing the sample, it needs to go through multiple steps, and the DNA in the sample There is loss, especially for high-concentration samples, insufficient lysis and incomplete enrichment will cause a large loss of DNA, resulting in sample loss verification, and low nucleic acid extraction efficiency; 2) Low detection sensitivity, around 5000copies / ml; 3) General There are no measures to prevent contamination; 4) There is no fluorescence normalization correction system, and the result judgment is greatly affected by subjective factors; 5) The timeliness of the methodology is poor and the throughput is small

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