QIAprep&amp Plasmodium Kit

For direct qPCR detection of Plasmodium spp. from dried blood spots or whole blood.

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QIAprep&amp Plasmodium Kit

Cat no. / ID.   223213

QIAprep&amp Plasmodium Kit is a qPCR master mix for direct detection of Plasmodium in liquid blood samples or in dried blood spot samples at a sensitivity of 1parasite/µL; for 100 reactions
The QIAprep&amp Plasmodium Kit is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention or treatment of a disease, nor has it been validated for such use either alone or in combination with other products.
Want to try this solution for the first time?
Get in touch with our specialists if you need a quote or if you would like to discuss your project.

Features

  • Detects Plasmodium DNA directly from dried blood spots or whole blood, saving time and reagent costs
  • Multiplex qPCR assays enable species differentiation (P. falciparum, P. vivax, P. malariae, P. ovale and P. knowlesi)
  • High sensitivity, detecting as low as 1 parasite/µL
  • Compatible with most real-time qPCR instruments

Product Details

The QIAprep&amp 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&amp 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. 

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Performance

The QIAprep&amp 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&amp Plasmodium Kit on DBS is either comparable or better than workflows with classic extraction 
      Hit Rate QIAprep&amp​ 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)

 

Principle

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.

Procedure

The QIAprep&amp 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&amp 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&amp 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&amp 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)

 

Applications

  • Malaria surveillance and epidemiological studies
  • Plasmodium detection in research labs
  • Plasmodium speciation in research labs
  • Field studies requiring fast, sensitive, extraction-free qPCR​

Supporting data and figures

FAQ

Which types of blood can be used in the Whole Blood and Sedimentation workflows?
Blood stabilized with EDTA, heparin, or citrate can be used.
FAQ ID - 4094
Is it possible to use reaction volumes smaller than 20 µL?
We do not recommend reducing the reaction volume, as this may cause inhibition and impact robustness of the assays.
FAQ ID - 4095
Can sensitivity be increased by using more sample (blood or dried blood spots)?
No, do not increase the amount of sample. All protocols are optimized to give highest sensitivity and robustness.
FAQ ID - 4096
Which types of blood cards can be used?
The QIAcard Bloodstain card (cat. no. WB100014), the QIAcard FTA Classic card (cat. no. WB 120205 or WB120305), and the Whatman filter cards (GE, cat. no. 3030 917 or 3017-915) have been successfully tested. We do not recommend FTA Elute cards due to their PCR inhibitory coating. Use Cutting Mat 2.5" x 3.0" (cat. no. WB100088) for punching.
FAQ ID - 4097
What are the recommended punch sizes for dried blood spot (DBS) samples?
We offer two DBS workflows, DBS Direct and DBS Elution.
For DBS Direct, one 1.2 mm punch is recommended; bigger or multiple punches may cause inhibition or interfere with signal detection.
For DBS Elution, one 3 mm punch is recommended; using multiple punches in our hands did not improve results.
For punching, UniCore Punch 1.2 mm (cat. no. WB100074) and UniCore Punch 3 mm (cat. no. WB100078) can be used.
FAQ ID - 4098
For dried blood spot (DBS) samples, when would I choose the Elution workflow vs the Direct workflow?
The Direct workflow offers a very fast process with very little hands-on and good sensitivity. The Elution workflow requires a bit more time and hands-on but provides higher sensitivity.
FAQ ID - 4099
Does it harm the amplification if, by accident, pieces of the cutting mat have been transferred along with the sample?
Depending on the applied pressure, pieces of the mat can be transferred into the reaction. This does not have a negative effect on the result, however, coring of the mat should be avoided.
FAQ ID - 4100
How should the puncher be cleaned between samples?
Incubate the puncher for 30 seconds each in 10% bleach, water, and 70% ethanol followed by an empty punch from a clean area of a card.
FAQ ID - 4101
For whole blood samples, when would I choose the Sedimentation workflow vs the Direct workflow?
The Direct workflow offers a quick process with minimal hands-on and good sensitivity.
The Sedimentation workflow requires a bit more process time and hands-on but provides higher sensitivity.
FAQ ID - 4102
Is the cell pellet formed in the Sedimentation workflow visible?
The pellet usually is not visible, which does not indicate an issue. We recommend arranging the tubes in a fixed orientation during centrifugation to collect the pellet at a defined spot.
FAQ ID - 4103
Is it possible to store the pellet lysed with buffer PR in the Sedimentation workflow for reuse?
Yes, it has been tested that the lysate can be stored at -20°C up to 4 weeks and 8 freeze-thaw cycles.
FAQ ID - 4104
How should the supernatant in the Sedimentation workflow be removed?
Carefully remove and discard the supernatant completely using a pipette. Do not decant the supernatant, as this causes significant cross contamination risks.
FAQ ID - 4105
What should I do if the Buffer PR looks cloudy?
Buffer PR can form a precipitate at temperatures below 15°C. We recommend equilibrating at a higher temperature until the buffer becomes clear. 
FAQ ID - 4107
Is it possible to store the S-solution at 4-8°C or room temperature?
We recommend storing the S-solution at -20°C protected from light. Short term storage at 4-8°C is possible, but we do not recommend room temperature storage.
FAQ ID - 4108