Description
The QIAprep& Plasmodium Kit streamlines Plasmodium detection by using direct qPCR. Researchers can analyze dried blood spots or whole blood directly, reducing hands-on time and reagent costs. The kit enables a high sensitivity of plasmodium detection at 1 parasite/µL, which is crucial for malaria surveillance and epidemiological research. Together with the QIAprep& Plasmodium assays, the differentiation of Plasmodium species (P. falciparum vs. non- falciparum to detection of P. vivax, P. malariae, P. ovale and P. knowlesi) takes place in one reaction in multiplex format. Would you like to learn more about the product from one of our specialists? Contact us, and we’ll get in touch with you shortly.
The QIAprep& Plasmodium Kit enables rapid qPCR analysis with results in 1–1.5 hours. It provides sensitivity comparable to or better than traditional DNA extraction-based methods, detecting down to 1 parasite/µL. The workflow is robust across different sample types and works with common real-time qPCR systems. The sensitivity of the QIAprep& Plasmodium Kit on DBS is either comparable or better than workflows with classic extraction Hit Rate QIAprep& Hit Rate classical DNA extraction followed by PCR Bloodstain card Classic card Samples Dilution Parasites per µL Liquid direct WF Liquid sedimentation WF DBS direct DBS elution DBS direct DBS elution Liquid DBS P. falciparum culture 1:100 79.4 100% 100% 100% 100% 100% 100% 100% 100% 1:1000 7.9 100% 100% 100% 100% 100% 100% 100% 100% 1:10,000 0.8 75% 100% 100% 100% 50% 100% 100% 50% 1:100,000 0.08 0% 88% 0% 88% 0% 75% 100% 25% Sample A 1:100 163.2 100% 100% 100% 100% 100% 100% 100% 100% 1:1000 16.3 100% 100% 100% 100% 100% 100% 100% 100% 1:10,000 1.6 88% 100% 100% 100% 83% 100% 100% 100% 1:100,000 0.16 50% 100% 33% 88% 50% 88% 100% 25% Sample B 1:100 159.9 100% 100% 100% 100% 100% 100% 100% 100% 1:1000 16.0 100% 100% 100% 100% 100% 100% 100% 100% 1:10,000 1.6 100% 100% 100% 100% 100% 88% 100% 75% 1:100,000 0.16 0% 100% 50% 75% 33% 38% 100% 0% P. falciparum culture and two human samples (Samples A and B) were diluted with Plasmodium -negative blood. DNA was extracted using the QIAamp® DNA Blood Mini Kit, and parasitemia was quantified via digital PCR with the QIAcuity® Probe PCR Kit on a QIAcuity Four Platform. For dried blood spot (DBS) workflows, 75 μL of diluted samples were applied to QIAcard Bloodstain or Classic card and dried for 24 h. Four workflows (Whole blood, sedmination, DBS direct and DBS elute) followed handbook protocols using the Pf/Non-Pf Detection Assay assay. Hit rates (DBS direct N=6; classical DNA workflows N=4; all others N=8) were recorded, with all samples run on Bio-Rad® CFX96™ or CFX384 instruments. (WF = workflow)
This kit integrates direct sample processing with qPCR to detect Plasmodium spp. For whole blood, a brief heat treatment releases nucleic acids. Dried blood spots can be used directly in the reaction or undergo minimal processing to recover eluate for use in several qPCR reactions with higher sensitivity. This kit is suitable for assays that detect Plasmodium DNA with high specificity and distinguish between P. falciparum and non- falciparum species. A second assay enables further species differentiation (P. vivax, P. malariae, P. ovale, and P. knowlesi). The two assays are available separately and can be used alone or in combination.
The QIAprep& Plasmodium Kit is suitable for whole blood samples and dried blood spots (DBS) as starting material. Two workflows are available for each sample type: ultrafast and ultrasensitive. Table 1. Selecting the right workflow according to your sample type and process requirements Sample Faster results in fewer steps More sensitive results Dried Blood Spots (DBS) DBS direct workflow DBS elution workflow Whole blood Whole blood workflow Sedimentation workflow Sample preparation The new QIAprep& Plasmodium Kit is designed for use with whole blood samples (for example, EDTA, citrate, heparin) and dried blood spots. Suitable sample types include: Fresh whole blood Frozen whole blood Dried blood spots from whole blood Dried blood spots from finger-prick blood The QIAprep& Plasmodium Kit technology allows the use of 20% of blood in the PCR reaction. Two workflows per sample type (whole blood or DBS) are provided. One workflow provides ultrafast results with few steps. An alternative workflow provides superior sensitivity with fewer handling steps (see Table 1). qPCR setup Combine the QIAprep& mastermix with the sample and assay mix. Load the reaction onto a PCR plate or vial and put it on a cycler. For more information, check the Quick Start Protocol or Product Handbook. Cycling and detection Run the real-time qPCR. Once the run is complete, analyze the results using the appropriate cycler software. For the generation of dried blood spots, we recommend the following: The QIAcard Bloodstain card (cat. no. WB100014) or the QIAcard FTA Classic card (cat. no. WB 120205 or WB120305) or Whatman filter cards Puncher, such as, UniCore Punch 1.2 mm for DBS direct WF (cat. no. WB100074), 3 mm for DBS Elution WF (cat. no. WB100078) or comparable punchers Cutting mat (cat. no. WB100088)
