RPB0560

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Plasmodium falciparum  Plasmodium falciparum,Plasmodium (Laverania) falciparum 5833 Haemosporida Plasmodiidae Plasmodium Plasmodium falciparum Eukaryota

Primer Description

Primer Name Sequence(5'-3') Length(bp) Primer Final Concentration(μM） GC Content（%） Predicted Melting Temperature(℃） Molecular Weight(g/moles) Positions in GenBank accession number
F Biotin-GCTTAGTTACGATTAATAGGAGTA 24 \ 33.33 48.34 7430.92 \
R Cy5-AGTCGGCATAGTTTATGGT 19 \ 42.11 50.09 5873.88 \

Gene Description

Target Gene GenBank ID
18S rRNA gene AF145334.1

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
The current duplex RPA methodology is highly specific and sensitive to detect multiple human Plasmodium spp. and is ready for point-of-need applications. The developed duplex RPA assays have potential applications in routine diagnostics and epidemiological studies. LF-RPA \ 20 min 37 °C LF 1 copy\µL 1 1

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2023 Lateral Flow Recombinase Polymerase Amplification Assays for the Detection of Human Plasmodium Species Meng Yee Lai,Mohd Abdul Hamid,Jenarun Jelip,Rose Nani Mudin,Yee Ling Lau The American journal of tropical medicine and hygiene 36913921 10.4269/ajtmh.22-0657

Lateral Flow Recombinase Polymerase Amplification Assays for the Detection of Human Plasmodium Species

Author(s):

Meng Yee Lai,Mohd Abdul Hamid,Jenarun Jelip,Rose Nani Mudin,Yee Ling Lau

Journal:

The American journal of tropical medicine and hygiene

Year:

2023

Abstract:

This study highlights the development of two lateral flow recombinase polymerase amplification assays for the diagnosis of human malaria. The lateral flow cassettes contained test lines that captured biotin-, 6-carboxyfluorescein, digoxigenin-, cyanine 5-, and dinitrophenyl-labeled amplicons. The overall process can be completed in 30 minutes. Recombinase polymerase amplification coupled with lateral flow had a detection limit of 1 copy/µL for Plasmodium knowlesi, Plasmodium vivax, and Plasmodium falciparum. No cross-reactivity was observed among nonhuman malaria parasites such as Plasmodium coatneyi, Plasmodium cynomolgi, Plasmodium brasilanium, Plasmodium inui, Plasmodium fragile, Toxoplasma gondii, Sarcocystis spp., Brugia spp., and 20 healthy donors. It is rapid, highly sensitive, robust, and easy to use. The result can be read without the need for special equipment and thus has the potential to serve as an effective alternative to polymerase chain reaction methods for the diagnosis of malaria.