Direct detection of equine herpesvirus type 1 DNA in nasal swabs by loop-mediated isothermal amplification (LAMP).
Abstract: We evaluated loop-mediated isothermal amplification (LAMP) as a means of detecting equine herpesvirus type 1 (EHV-1) DNA directly from nasal swabs. To increase the sensitivity, we added a step in which the samples were heat-treated to the original LAMP procedure. The detection limit of the LAMP assay with heat treatment was 10 times more sensitive than the original LAMP assay even when the DNA extraction step was omitted. In addition, the LAMP assay with heat treatment was more sensitive than the original LAMP assay and the polymerase chain reaction using clinical samples. The LAMP assay with heat treatment is easy to perform and so should be applicable to the diagnosis of EHV-1 infections in clinical laboratories.
Publication Date: 2011-05-06 PubMed ID: 21551979DOI: 10.1292/jvms.11-0065Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
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 research article focuses on an evaluation of the loop-mediated isothermal amplification (LAMP) method for detecting equine herpesvirus type 1 (EHV-1) DNA directly from nasal swabs in horses, with changes made to sensitize it further.
Loop-Mediated Isothermal Amplification (LAMP)
- The core technique used in this study for the detection of EHV-1 is Loop-Mediated Isothermal Amplification (LAMP). It is a nucleic acid amplification method designed to amplify specific DNA sequences in a highly efficient and rapid process.
- The LAMP method is found to be a viable tool due to its simplicity, speed, and high specificity in targeting the DNA of interest.
Adapted LAMP with Heat Treatment
- The researchers revised the original LAMP procedure by incorporating an additional heat treatment step. The heat treatment of samples is a known method to increase the sensitivity of various assays, leading to more accurate results.
- Upon implementing this adapted LAMP process, the researchers found that its detection limit increased up to ten times more than the original LAMP method. This dramatic improvement in sensitivity was observed even when the DNA extraction step was excluded.
Comparison with Other Processes
- The study compared this revised LAMP method with the original LAMP technique and the standard polymerase chain reaction (PCR), commonly used for detecting and amplifying specific DNA sequences.
- Results showed that the adapted LAMP method was more effective for detecting EHV-1 in clinical samples than both the original LAMP process and PCR, underlining its potential for use in practical applications.
Potential Applications
- Due to its higher sensitivity, simplicity, and ease of performance, the adapted LAMP method could find widespread applications in diagnostic laboratories. It has the potential to become a new standard for detection and diagnosis of EHV-1 infections in horses.
Cite This Article
APA
Nemoto M, Ohta M, Tsujimura K, Bannai H, Yamanaka T, Kondo T, Matsumura T.
(2011).
Direct detection of equine herpesvirus type 1 DNA in nasal swabs by loop-mediated isothermal amplification (LAMP).
J Vet Med Sci, 73(9), 1225-1227.
https://doi.org/10.1292/jvms.11-0065 Publication
Researcher Affiliations
- Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400–4 Shiba, Shimotsuke, Tochigi 329–0412, Japan. nemoto_manabu@epizoo.equinst.go.jp
MeSH Terms
- Animals
- DNA, Viral / isolation & purification
- Herpesviridae Infections / diagnosis
- Herpesviridae Infections / veterinary
- Herpesviridae Infections / virology
- Herpesvirus 1, Equid / genetics
- Herpesvirus 1, Equid / isolation & purification
- Horse Diseases / diagnosis
- Horse Diseases / virology
- Horses
- Mucus / virology
- Nucleic Acid Amplification Techniques / veterinary
- Sensitivity and Specificity
Citations
This article has been cited 6 times.- Takehana K, Matsuno K. Direct detection of elephant endotheliotropic herpesvirus 1 (EEHV1) DNA in heparinized plasma by loop-mediated isothermal amplification.. J Vet Med Sci 2023 Mar 30;85(4):459-462.
- Jelocnik M, Nyari S, Anstey S, Playford N, Fraser TA, Mitchell K, Blishen A, Pollak NM, Carrick J, Chicken C, Jenkins C. Real-time fluorometric and end-point colorimetric isothermal assays for detection of equine pathogens C. psittaci and equine herpes virus 1: validation, comparison and application at the point of care.. BMC Vet Res 2021 Aug 19;17(1):279.
- Kinoshita Y, Niwa H, Katayama Y. Development of a loop-mediated isothermal amplification method for detecting Streptococcus equi subsp. zooepidemicus and analysis of its use with three simple methods of extracting DNA from equine respiratory tract specimens.. J Vet Med Sci 2014 Sep;76(9):1271-5.
- Malik YS, Sharma K, Kumar N, Shivachandra SB, Rawat V, Rakholia R, Ranjan R, Ganesh B, Parida M. Rapid detection of human rotavirus using NSP4 gene specific reverse transcription loop-mediated isothermal amplification assay.. Indian J Virol 2013 Sep;24(2):265-71.
- Nie K, Qi SX, Zhang Y, Luo L, Xie Y, Yang MJ, Zhang Y, Li J, Shen H, Li Q, Ma XJ. Evaluation of a direct reverse transcription loop-mediated isothermal amplification method without RNA extraction for the detection of human enterovirus 71 subgenotype C4 in nasopharyngeal swab specimens.. PLoS One 2012;7(12):e52486.
- Mukhopadhyay HK, Amsaveni S, Matta SL, Antony PX, Thanislass J, Pillai RM. Development and evaluation of loop-mediated isothermal amplification assay for rapid and sensitive detection of canine parvovirus DNA directly in faecal specimens.. Lett Appl Microbiol 2012 Sep;55(3):202-9.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
