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Evaluation of three rapid low-resource molecular tests for Nipah virus
Journal article   Open access   Peer reviewed

Evaluation of three rapid low-resource molecular tests for Nipah virus

Nina Pollak, Malin Olsson, Glenn Marsh, Joanne Macdonald and David McMillan
Frontiers in Microbiology, Vol.13, pp.1-12
2023
PMCID: PMC9949527
PMID: 36845977
pdf
fmicb-13-11019143.30 MBDownloadView
Published Version Open Access CC BY V4.0
url
https://doi.org/10.3389/fmicb.2022.1101914View
Published Version

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Abstract

Nipah virus (NiV) rapid test isothermal amplification nucleic acid lateral flow point of care recombinase polymerase amplification (RPA) recombinase-aided amplification (RAA0 nucleic acid extraction
Accurate and timely diagnosis of Nipah virus (NiV) requires rapid, inexpensive, and robust diagnostic tests to control spread of disease. Current state of the art technologies are slow and require laboratory infrastructure that may not be available in all endemic settings. Here we report the development and comparison of three rapid NiV molecular diagnostic tests based on reverse transcription recombinase-based isothermal amplification coupled with lateral flow detection. These tests include a simple and fast one-step sample processing step that inactivates the BSL-4 pathogen, enabling safe testing without the need for multi-step RNA purification. The rapid NiV tests targeted the Nucleocapsid protein (N) gene with analytical sensitivity down to 1,000 copies/μL for synthetic NiV RNA and did not cross-react with RNA of other flaviviruses or Chikungunya virus, which can clinically present with similar febrile symptoms. Two tests detected 50,000–100,000 TCID50/mL (100–200 RNA copies/reaction) of the two distinct strains of NiV, Bangladesh (NiVB) and Malaysia (NiVM), and took 30 min from sample to result, suggesting these tests are well suited for rapid diagnosis under resource-limited conditions due to rapidity, simplicity, and low equipment requirements. These Nipah tests represent a first step toward development of near-patient NiV diagnostics that are appropriately sensitive for first-line screening, sufficiently robust for a range of peripheral settings, with potential to be safely performed outside of biohazard containment facilities.

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