PCR Malaria Detection Oligonucleotide Primers
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Solution Overview
Problem
There is a need for a quick, reliable, specific, and sensitive method for detecting and quantifying the presence of Malaria parasites in biological samples, particularly for blood donations, as existing methods like microscopy, RDTs, and enzyme immunoassays are not sufficient for high-throughput and sensitive detection.
Innovation Solution
A polymerase chain reaction (PCR)-based assay using specific oligonucleotide primers and probes, such as those with sequences SEQ ID NOs: 1-15, for detecting and quantitating Malaria parasites like Plasmodium species in a single test tube, employing real-time PCR and fluorescent resonance energy transfer (FRET) for detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If microscopy or RDTs are used for Malaria detection, then the method is simple and inexpensive, but the sensitivity and throughput are insufficient
Solution Approach 1:
The patent replaces mechanical microscopy methods with a molecular biology-based PCR system. The detection mechanism transitions from visual examination of stained blood smears to amplification and detection of parasite DNA using oligonucleotide primers and probes, achieving higher sensitivity through biochemical amplification rather than mechanical observation
Solution Approach 2:
The patent changes the detection parameter from visual/immunological detection to nucleic acid amplification detection. By targeting specific DNA sequences of Plasmodium species and using fluorescently labeled probes, the system achieves quantitative detection with much lower limits of detection compared to traditional methods
2Reliability
If donor deferral policy is implemented, then blood supply safety is improved, but blood supply quantity decreases
Solution Approach 1:
The patent implements preliminary screening of blood donations using highly sensitive PCR-based Malaria detection before transfusion. By detecting Malaria parasites at very low levels in donated blood, the system identifies infected donations early, allowing for targeted deferral of only those specific donations rather than blanket deferral policies
Solution Approach 2:
The patent uses nucleic acid amplification to create multiple copies of the parasite DNA present in the blood sample. This amplification process enables detection of extremely low levels of Malaria parasites that would otherwise be undetectable, allowing for more accurate identification of truly safe donations
3Quantity of substance
If selective testing is implemented, then impact on blood supply is reduced, but detection sensitivity must be maintained
Solution Approach 1:
The patent replaces less sensitive immunological testing methods with molecular PCR-based detection. This substitution maintains high detection sensitivity while allowing selective testing of only those donations that require it, rather than universal deferral or testing policies
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The PCR-based assay provides rapid, accurate, and sensitive detection and quantitation of Malaria parasites, ensuring the safety of blood supplies by identifying Plasmodium species with high specificity and sensitivity, even at low parasite levels.
Implementation Method 1
employing real-time PCR and fluorescent resonance energy transfer (FRET) for detection
Data Source
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AI summary
Methods for the rapid detection of the presence or absence of Malaria parasites (including Plasmodium) in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Additionally, the assay can be a multiplex assay, to amplify and detect a plurality of Plasmodium targets simultaneously, which offers advantages over singleplex assays. Furthermore, primers and probes targeting Malaria parasites (including Plasmodium) and kits are provided that are designed for the detection of Plasmodium, including, but not limited to, the Plasmodium species of P. falciparum, P. vivax, P. ovale, P. knowlesi, and P. malariae. Also described are kits, reaction mixtures, and oligonucleotides (e.g., primer and probe) for the amplification and detection of Malaria parasites (including Plasmodium).