Quickly-nested PCR (polymerase chain reaction) malaria molecular diagnosis reagent kit and detection method
A technology of molecular detection and reagent kit, which is applied in the field of medical testing, can solve the problems of complex reagent operation, easy pollution, and long test time, and achieve the effects of simplifying reagents and operating steps, improving sensitivity, and reducing costs
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Embodiment 1
[0070] Example 1: Detection of Plasmodium falciparum and Plasmodium vivax using a rapid nested PCR malaria molecular diagnostic kit——sensitivity detection, see the results figure 2 .
[0071] Preparation of blood samples from blood filter papers simulated by density gradient infection of Plasmodium: collect one part of whole blood containing parasites from patients diagnosed with simple Plasmodium falciparum and simple Plasmodium vivax infection (above the density of protozoa 10000 worms / μl of blood) from veins, and use healthy Human anticoagulated whole blood was diluted with 10-fold serial dilution method to obtain 10000 / μl, 1000 / μl, 100 / μl, 10 / μl, 1 / μl gradient worm-containing blood, and use A pipette was used to quantitatively prepare a blood sample of Plasmodium-containing blood filter paper with a density gradient of plasmodia (5 μl / per piece of 6mm diameter filter paper disc), and after natural air drying, it was divided into sealed tubes and stored at 4°C for later us...
Embodiment 2
[0086] Example 2: Detection of Plasmodium falciparum and Plasmodium vivax by using the Rapid Nested PCR Malaria Molecular Diagnosis Kit - Detecting blood samples of malaria febrile patients in malaria surveillance, see the results image 3 .
[0087] Blood sample collection: Use the filter paper method to collect blood samples from clinically diagnosed malaria or suspected malaria patients. Routinely disinfected earlobe or fingertip blood pricking can be used. The center of the filter paper is directly in contact with the blood drop, so that the blood penetrates the filter paper to a diameter of 2-4cm. It is used after natural air drying. Put the white paper in a film bag at intervals and mail it at room temperature or store it at 4°C; or use a loose-leaf paper puncher to punch out small discs with a diameter of 6mm (containing about 4-5μl whole blood), and put them in a 0.5-1.5ml conical tube 4 Store at ℃.
[0088] Preparation of filter paper blood sample DNA template: use a...
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