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Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay for the Specific and Rapid Detection of Tilapia Lake Virus
Published on: May 18, 2020
Specific and quantitative assay for duck variant orthoreovirus utilizing the loop-mediated isothermal amplification
Shuai Zhang1, Xinhui Wei2, Min Han3
1Jinan Vocational College of Nursing, Jinan, Shandong Province 250104, China; College of Veterinary Medicine, Shandong Agricultural University, Tai'an, Shandong Province 271018, China.
Journal of Immunological Methods
|May 21, 2026
Summary
A new quantitative loop-mediated isothermal amplification assay (qLAMP) offers a sensitive and specific method for detecting novel avian orthoreovirus (NDRV) in ducks. This rapid diagnostic tool aids in managing NDRV infections and preventing disease spread.
Area of Science:
- Veterinary Virology
- Molecular Diagnostics
- Avian Pathology
Background:
- Novel avian orthoreovirus (NDRV) strains have caused significant illness and splenic necrosis in ducks since 2018.
- Accurate and rapid diagnostic methods are crucial for managing avian reovirus (ARV) infections.
Purpose of the Study:
- To develop a novel quantitative loop-mediated isothermal amplification (qLAMP) assay for the detection of NDRV.
- To evaluate the sensitivity, specificity, and reproducibility of the developed qLAMP assay.
Main Methods:
- Development of a unique qLAMP assay for NDRV identification.
- Testing the assay with serial dilutions of standard substance to determine detection limits.
- Assessing specificity through cross-reaction tests with other avian viruses.
- Evaluating reproducibility using intra- and interassay variability tests.
- Comparing qLAMP efficiency with traditional PCR and RT-qPCR using clinical samples.
Main Results:
- The qLAMP assay achieved a low detection limit of 1.0 copies/μL.
- The assay demonstrated high specificity, with no cross-reaction with other avian viruses.
- Excellent reproducibility was confirmed with variability ranging from 0.08% to 2.78%.
- qLAMP showed greater detection efficiency compared to PCR and RT-qPCR in 90 clinical samples.
Conclusions:
- The developed qLAMP assay is a sensitive and specific tool for identifying NDRV in clinical samples from various avian species.
- This assay can serve as a standard diagnostic test for NDRV detection in laboratories.
- The qLAMP assay facilitates timely diagnosis and management of NDRV outbreaks in poultry.
