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Journal of virological methods2015; 215-216; 13-16; doi: 10.1016/j.jviromet.2015.02.001

Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification.

Abstract: A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the rapid detection of equine coronavirus (ECoV). This assay was conducted at 60 °C for 40 min. Specificity of the RT-LAMP assay was confirmed using several equine intestinal and respiratory pathogens in addition to ECoV. The novel assay failed to cross-react with the other pathogens tested, suggesting it is highly specific for ECoV. Using artificially synthesized ECoV RNA, the 50% detection limit of the RT-LAMP assay was 10(1.8)copies/reaction. This is a 50-fold greater sensitivity than conventional reverse transcription polymerase chain reaction (RT-PCR) assays, but a 4-fold lower sensitivity than quantitative RT-PCR (qRT-PCR) assays. Eighty-two fecal samples collected during ECoV outbreaks were analyzed. ECoV was detected in 59 samples using the RT-LAMP assay, and in 30 and 65 samples using RT-PCR or qRT-PCR assays, respectively. Although the RT-LAMP assay is less sensitive than qRT-PCR techniques, it can be performed without the need for expensive equipment. Thus, the RT-LAMP assay might be suitable for large-scale surveillance and diagnosis of ECoV infection in laboratories with limited resources.
Copyright © 2015 Elsevier B.V. All rights reserved.
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Publication Date: 2015-02-12 PubMed ID: 25682750PubMed Central: PMC7113660DOI: 10.1016/j.jviromet.2015.02.001Google Scholar: Lookup
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  • Comparative Study
  • Evaluation Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study is about a new diagnostic method for fast detection of equine coronavirus, called RT-LAMP, which showed a high level of specificity and more sensitivity than traditional tests, allowing for easier, cheaper large-scale monitoring of the virus.

Overview of Research

  • This research focuses on the development of a new diagnostic technique known as Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for the quick detection of Equine Coronavirus (ECoV).
  • The technique was tested at 60 degrees Celsius over a 40-minute period.

Specificity of the RT-LAMP Assay

  • The RT-LAMP assay was evaluated for its specificity against ECoV by testing it on several equine intestinal and respiratory pathogens.
  • Results showed that this assay failed to cross-react with other pathogens tested, indicating that it is very precise in identifying ECoV.

Sensitivity of the RT-LAMP Assay

  • The researchers also tested the sensitivity of the RT-LAMP assay using artificially synthesized ECoV RNA.
  • The 50% detection limit of this assay was found to be 10(1.8) copies/reaction, showing a 50-fold greater sensitivity than conventional Reverse Transcription Polymerase Chain Reaction (RT-PCR) assays, but 4-fold less sensitive than Quantitative RT-PCR (qRT-PCR) assays.

Comparison with Other Tests

  • In comparison to RT-PCR and qRT-PCR assays on 82 fecal samples collected from ECoV outbreaks, the RT-LAMP assay detected 59 samples to be positive with ECoV.
  • Whereas, RT-PCR and qRT-PCR assays identified 30 and 65 samples respectively to be positive with ECoV, indicating lower sensitivity for RT-PCR and higher sensitivity for qRT-PCR relative to RT-LAMP.

Benefits of the RT-LAMP Assay

  • Despite the RT-LAMP assay being less sensitive than qRT-PCR techniques, it can be conducted without the need for costly equipment.
  • As a result, the RT-LAMP assay might be suitable for large-scale surveillance and diagnosis of ECoV infection, specifically in laboratories with limited financial resources.

Cite This Article

APA
Nemoto M, Morita Y, Niwa H, Bannai H, Tsujimura K, Yamanaka T, Kondo T. (2015). Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification. J Virol Methods, 215-216, 13-16. https://doi.org/10.1016/j.jviromet.2015.02.001

Publication

Journal of virological methods
ISSN: 1879-0984
NlmUniqueID: 8005839
Country: Netherlands
Language: English
Volume: 215-216
Pages: 13-16
PII: S0166-0934(15)00023-3

Researcher Affiliations

Nemoto, Manabu
  • Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan. Electronic address: nemoto_manabu@epizoo.equinst.go.jp.
Morita, Yoshinori
  • Tokachi Draft Horse Clinic, Nishi 13, Minami 9-1, Obihiro, Hokkaido 080-0023, Japan.
Niwa, Hidekazu
  • Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan.
Bannai, Hiroshi
  • Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan.
Tsujimura, Koji
  • Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan.
Yamanaka, Takashi
  • Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan.
Kondo, Takashi
  • Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan.

MeSH Terms

  • Animals
  • Coronavirus / isolation & purification
  • Coronavirus Infections / diagnosis
  • Coronavirus Infections / virology
  • Cross Reactions
  • Feces / virology
  • Horse Diseases / diagnosis
  • Horse Diseases / virology
  • Horses
  • Molecular Diagnostic Techniques / methods
  • Nucleic Acid Amplification Techniques / methods
  • Sensitivity and Specificity
  • Temperature
  • Time Factors

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Citations

This article has been cited 11 times.
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