Rapid and high-throughput plasmodium detection method

A detection method and technology for Plasmodium, applied in the field of rapid and high-throughput detection of Plasmodium, can solve problems such as complex operations, and achieve the effect of improving detection capability, reducing time and complexity

Inactive Publication Date: 2015-04-29
SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The current gene chip detection for malaria parasite detection has been used in some researches, but the probe spotting of these gene chips mostly adopts manual mode, which is complicated to operate and increases the chance of many false positives

Method used

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  • Rapid and high-throughput plasmodium detection method
  • Rapid and high-throughput plasmodium detection method
  • Rapid and high-throughput plasmodium detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Specificity and sensitivity of embodiment 1 malaria parasite detection

[0066] 1. Experimental steps

[0067] 1. Collection of blood samples

[0068] Take 1.0ml of whole blood sample from the patient at the port for testing.

[0069] 2. Preliminary Identification of Blood Samples

[0070] The colloidal gold method was used to preliminarily determine whether the patient was infected with Plasmodium.

[0071] 3. Extraction of Genomic DNA

[0072] Plasmodium DNA was extracted using a blood sample genomic DNA extraction kit (spin column type, Qiagen). Take 50uL whole blood sample, and extract DNA according to the extraction steps in the kit manual.

[0073] 4. Determination of DNA concentration

[0074] The concentration of Plasmodium DNA was quantified by real-time quantitative PCR using ABI's real-time fluorescence detector.

[0075] 5. Amplification of target genes

[0076] 5.1 Design of primers and probes

[0077] Aiming at the unique genes of Plasmodium, desi...

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Abstract

The invention relates to a rapid and high-throughput plasmodium detection method. According to the method disclosed by the invention, two powerful molecular biological technologies of polymerase chain reaction (PCR) and micro-array are integrated, a PCR hybridization probe is directly fixed in a hybridization cabin and arranged on the same chip as a PCR reaction chamber; the detection method comprises the steps of blood sample DNA liquid extraction, PCR amplification, hybridization, washing and result analysis. The rapid and high-throughput plasmodium detection method disclosed by the invention can be used for rapid sensitive detection of falciparum malaria plasmodium, vivax malaria plasmodium and quartan malaria plasmodium, the detection efficiency of front-line inspection and quarantine personnel at import-export ports is greatly improved, not only is the workload reduced but also the problem of positive undetected samples possibly missed according to the conventional detection methods can be solved to the largest extent, and therefore the occurrence of plasmodium epidemic situation is prevented to the largest extent.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnostic reagents, in particular to a rapid and high-throughput detection method for malaria parasites. Background technique [0002] Malaria is one of the most serious global public health problems, and more than half of the world's population lives in malaria-endemic areas. In recent years, imported malaria in my country has been on the rise year by year. There are four species of Plasmodium that infect humans: Plasmodium falciparum (P.f), Plasmodium vivax (P.v), Plasmodium malariae (P.m ) and Plasmodium ovale (P.o). Therefore, effective detection of Plasmodium is an essential means for medical diagnosis and malaria control. At present, microscopic examination is still used as the "gold standard" for diagnosing malaria in my country. Microscopic examination is affected by the subjective judgment and sense of responsibility of microscopic personnel, and it is very easy to miss diagnosis and d...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6837C12Q2531/113C12Q2565/501Y02A50/30
Inventor 田桢干张子龙李深伟李美
Owner SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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