Qiagen, 223213, QIAprep& Plasmodium Kit

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

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

Hit Rate QIAprep&amp​: Hit Rate classical DNA extraction followed by PCR​
Bloodstain card​: Classic card​
Samples​: Dilution​
P. falciparum culture​: 1:100​
1:1000​: 7.9​
1:10,000​: 0.8​
1:100,000​: 0.08​
Sample A​: 1:100​
1:1000​: 16.3​
1:10,000​: 1.6​
1:100,000​: 0.16​
Sample B​: 1:100
1:1000​: 16.0​
1:10,000: 1.6​
1:100,000​: 0.16

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​​.

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

Sample: Faster results in fewer steps
Dried Blood Spots (DBS): DBS direct workflow
Whole blood: Whole blood workflow

- Fresh whole blood
- Frozen whole blood
- Dried blood spots from whole blood
- Dried blood spots from finger-prick blood

- 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)

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 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:

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:

Applications

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