Development of a recombinase polymerase amplification (RPA-EXO) and lateral flow assay (RPA-LFA) based on the ITS1 gene for the detection of Angiostrongylus cantonensis in gastropod intermediate hosts
| Autor: | Argon Steel, Elizabeth S Atkinson, Susan I Jarvi, Lisa Kaluna, Kirsten Snook |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Serial dilution 030231 tropical medicine Recombinase Polymerase Amplification Biology biology.organism_classification Molecular biology Angiostrongylus cantonensis law.invention 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology 0302 clinical medicine Infectious Diseases Real-time polymerase chain reaction Plasmid chemistry law Animal Science and Zoology Parasitology Gene DNA Polymerase chain reaction |
| Zdroj: | Parasitology. 148:251-258 |
| ISSN: | 1469-8161 0031-1820 |
| DOI: | 10.1017/s0031182020002139 |
| Popis: | Angiostrongylus cantonensis is a parasitic nematode known to infect humans through the ingestion of third stage larvae which can cause inflammation and damage to the central nervous system. Currently, polymerase chain reaction (PCR) is one of the most reliable diagnostic methods for detecting A. cantonensis in humans as well as in gastropod hosts, but requires expensive and specialized equipment. Here, we compare the sensitivity and accuracy of a recombinase polymerase amplification Exo (RPA-EXO) assay, and a recombinase polymerase amplification lateral flow assay (RPA-LFA) with a traditional quantitative PCR (qPCR) assay currently available. The three assays were used to test 35 slugs from Hawai‘i for the presence of A. cantonensis DNA. Consistent results among the three tests were shown in 23/35 samples (65.7%), while 7/35 (20%) were discordant in low infection level samples (μL−1 to ~1 copy μL−1. All three assays consistently detected 50–100 copies μL−1 in triplicate and qPCR was able to detect ~13 copies μL−1 in triplicate. RPA-EXO was able to detect 25 copies μL−1 in triplicate and RPA-LFA was not able to amplify consistently below 50 copies μL−1. Thus, our RPA-EXO and RPA-LFA assays do not appear as sensitive as the current qPCR assay at low DNA concentrations; however, these tests have numerous advantages that may make them useful alternatives to qPCR. |
| Databáze: | OpenAIRE |
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