Recombinase Polymerase Amplification Compared to Real-Time Polymerase Chain Reaction Test for the Detection of Fasciola hepatica in Human Stool
| Autor: | Hayley Rogers, Miguel M. Cabada, Maryann I Mbaka, A. Clinton White, Patrick A. Naeger, Alejandro Castellanos-Gonzalez, Kelli A. Bagwell, Safa Maharsi, Jose L. Malaga |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Specific test 030231 tropical medicine Recombinase Polymerase Amplification Real-Time Polymerase Chain Reaction/methods complex mixtures Sensitivity and Specificity law.invention 03 medical and health sciences 0302 clinical medicine law Virology parasitic diseases Fasciola hepatica Humans Animals Fasciola hepatica/genetics Nucleic Acid Amplification Techniques/methods Polymerase chain reaction Microscopy Thermal cycler Fasciola biology Feces/parasitology biology.organism_classification Molecular biology Highly sensitive enzymes and coenzymes (carbohydrates) 030104 developmental biology Infectious Diseases Real-time polymerase chain reaction Fascioliasis/diagnosis/parasitology Parasitology purl.org/pe-repo/ocde/ford#3.03.06 [https] |
| Popis: | Fasciola hepatica is the most widely distributed trematode infection in the world. Control efforts may be hindered by the lack of diagnostic capacity especially in remote endemic areas. Polymerase chain reaction (PCR)–based methods offer high sensitivity and specificity but require expensive technology. However, the recombinase polymerase amplification (RPA) is an efficient isothermal method that eliminates the need for a thermal cycler and has a high deployment potential to resource-limited settings. We report on the characterization of RPA and PCR tests to detect Fasciola infection in clinical stool samples with low egg burdens. The sensitivity of the RPA and PCR were 87% and 66%, respectively. Both tests were 100% specific showing no cross-reactivity with trematode, cestode, or nematode parasites. In addition, RPA and PCR were able to detect 47% and 26% of infections not detected by microscopy, respectively. The RPA adapted to a lateral flow platform was more sensitive than gel-based detection of the reaction products. In conclusion, the Fasciola RPA is a highly sensitive and specific test to diagnose chronic infection using stool samples. The Fasciola RPA lateral flow has the potential for deployment to endemic areas after further characterization. |
| Databáze: | OpenAIRE |
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