Hendra virus detection using Loop-Mediated Isothermal Amplification.
Abstract: Hendra virus (HeV) is a zoonotic paramyxovirus endemic in Australian Pteropus bats (fruit bats or flying foxes). Although bats appear to be unaffected by the virus, HeV can spread from fruit bats to horses, causing severe disease. Human infection results from close contact with the blood, body fluids and tissues of infected horses. HeV is a biosecurity level 4 (BSL-4) pathogen, with a high case-fatality rate in humans and horses. Current assays for HeV detection require complex instrumentation and are generally time consuming. The aim of this study was to develop a Loop-Mediated Isothermal Amplification (LAMP) assay to detect nucleic acid from all known HeV strains in horses without the requirement for complex laboratory equipment. A LAMP assay targeting a conserved region of the HeV P-gene was combined with a Lateral Flow Device (LFD) for detection of amplified product. All HeV isolates, the original HeV isolated in 1994 as well as the most recent isolates from 2011 were detected. Analytical sensitivity and specificity of the HeV-LAMP assay was equal to a TaqMan assay developed previously. Significantly, these assays detected HeV in horses before clinical signs were observed. The combined LAMP-LFD procedure is a sensitive method suitable for HeV diagnosis in a resource-limited situation or where rapid test results are critical.
Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.
Publication Date: 2012-02-02 PubMed ID: 22327143DOI: 10.1016/j.jviromet.2012.01.020Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Biosecurity
- Diagnosis
- Diagnostic Technique
- Disease control
- Disease Diagnosis
- Disease Management
- Disease Outbreaks
- Disease Surveillance
- Disease Transmission
- Disease Treatment
- Epidemiology
- Equine Health
- Hendra Virus
- Horses
- Infectious Disease
- Laboratory Methods
- Public Health
- Veterinary Medicine
- Veterinary Science
- Virology
- Zoonotic Diseases
Summary
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This research focuses on the development of a fast and less complex method for detecting the Hendra virus (HeV) in horses, using a technique called Loop-Mediated Isothermal Amplification (LAMP).
Introduction and Background
- HeV is an infectious and deadly virus endemic to fruit bats in Australia. Though harmless to bats, the virus can spread to horses and possibly to humans upon contact with contaminated blood, body fluids, and tissues from the infected horses.
- Current assays for HeV detection are usually time-consuming and require complex equipment and techniques.
- HeV is categorized as a Biosecurity Level 4 (BSL-4) pathogen, which refers to a high-risk group of biological agents, emphasizing the urgency to develop efficient and quick detection methods.
Aim of the Study and Methodology
- The primary goal of the study was to establish a LAMP assay that can detect HeV strains in horses without the need for intricate laboratory equipment.
- The researchers aimed at targeting a conserved region of the HeV P-gene using the LAMP assay. The amplified product was then detected using a Lateral Flow Device (LFD).
- Hendra Virus isolates from 1994 up to 2011 were tested using this method.
Key Findings
- The newly developed LAMP assay showed equal sensitivity and specificity to a previously developed TaqMan assay, a PCR-based technique used for detecting and quantifying specific DNA sequences.
- This method managed to detect the presence of HeV in horses even before any clinical signs were evident. This early detection capability is significant in preventing potential outbreaks and facilitating early treatment.
- The combination of LAMP and LFD is a promising method for diagnosing HeV, especially in settings where resources are limited or when results are urgently needed.
Cite This Article
APA
Foord AJ, Middleton D, Heine HG.
(2012).
Hendra virus detection using Loop-Mediated Isothermal Amplification.
J Virol Methods, 181(1), 93-96.
https://doi.org/10.1016/j.jviromet.2012.01.020 Publication
Researcher Affiliations
- CSIRO Australian Animal Health Laboratory, 5 Portarlington Road, Geelong, VIC 3220, Australia.
MeSH Terms
- Animals
- Australia
- Hendra Virus / isolation & purification
- Henipavirus Infections / diagnosis
- Henipavirus Infections / veterinary
- Horse Diseases / diagnosis
- Horse Diseases / virology
- Horses
- Molecular Diagnostic Techniques / methods
- Nucleic Acid Amplification Techniques / methods
- Sensitivity and Specificity
- Virology / methods
Citations
This article has been cited 9 times.- van den Hurk S, Yondo A, Velayudhan BT. Laboratory Diagnosis of Hendra and Nipah: Two Emerging Zoonotic Diseases with One Health Significance. Viruses 2025 Jul 17;17(7).
- Hulme PE, Beggs JR, Binny RN, Bray JP, Cogger N, Dhami MK, Finlay-Smits SC, French NP, Grant A, Hewitt CL, Jones EE, Lester PJ, Lockhart PJ. Emerging advances in biosecurity to underpin human, animal, plant, and ecosystem health. iScience 2023 Sep 15;26(9):107462.
- Pollak NM, Marsh GA, Olsson M, McMillan D, Macdonald J. Rapid, sensitive, and specific, low-resource molecular detection of Hendra virus. One Health 2023 Jun;16:100504.
- Knox A, Beddoe T. Isothermal Nucleic Acid Amplification Technologies for the Detection of Equine Viral Pathogens. Animals (Basel) 2021 Jul 20;11(7).
- Li J, Liang W, Xu S, Shi J, Zhou X, Liu B, Yu L, Xiong J, Si G, He D. Rapid and sensitive detection of Senecavirus A by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick method. PLoS One 2019;14(5):e0216245.
- Ma L, Chen Z, Guan W, Chen Q, Liu D. Rapid and Specific Detection of All Known Nipah virus Strains' Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification. Front Microbiol 2019;10:418.
- Zhuang L, Ji Y, Tian P, Wang K, Kou C, Gu N, Zhang Y. Polymerase chain reaction combined with fluorescent lateral flow immunoassay based on magnetic purification for rapid detection of canine parvovirus 2. BMC Vet Res 2019 Jan 17;15(1):30.
- Ge Y, Wu B, Qi X, Zhao K, Guo X, Zhu Y, Qi Y, Shi Z, Zhou M, Wang H, Cui L. Rapid and sensitive detection of novel avian-origin influenza A (H7N9) virus by reverse transcription loop-mediated isothermal amplification combined with a lateral-flow device. PLoS One 2013;8(8):e69941.
- Foord AJ, White JR, Colling A, Heine HG. Microsphere suspension array assays for detection and differentiation of Hendra and Nipah viruses. Biomed Res Int 2013;2013:289295.
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