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Updated: Jan 3, 2026
Author Spotlight: Advancing Pathogen Diagnostics with Standardized LAMP
Published on: September 8, 2023
Rapid and sensitive detection of shrimp infectious myonecrosis virus using a reverse transcription loop-mediated
Narong Arunrut1, Jantana Kampeera, Rungkarn Suebsing
1Center of Excellence for Shrimp Molecular Biology and Biotechnology (CENTEX Shrimp), Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand; National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathumthani 12120, Thailand.
Abstract:
This study reports a novel strategy for the detection of reverse transcription loop-mediated isothermal amplification (RT-LAMP) products derived from infectious myonecrosis virus (IMNV), causes a serious myonecrosis in Penaeus (Litopenaeus) vannamei, by using a ssDNA-labeled with gold nanoparticle (AuNP) probe. This technique relies on a self-aggregation method, when the AuNP aggregation is induced by an increasing of salt concentrations with visual detection. The presence of IMNV-LAMP target prevented an AuNP aggregation and a solution remained as pink color of AuNP, while non-complementary targets cannot prevent AuNP aggregation, resulting in a visible color change to purple color after addition of salt. By using the combination of LAMP and AuNP probe system, the total assay interval required approximately 50 min (exclude RNA preparation). Detection limit was 10 copies of IMNV RNA in vitro transcript that comparable to that of LAMP followed by LFD and nested RT-PCR, but it was 100-times more sensitive than RT-PCR methods. This assay can be adapted easily for rapid detection of other shrimp infectious diseases agents at low-cost with robust reagents and using a simple colorimetric detection method.
Insights
A new method detects infectious myonecrosis virus (IMNV) using gold nanoparticles and reverse transcription loop-mediated isothermal amplification (RT-LAMP). This rapid, sensitive colorimetric assay offers a low-cost solution for shrimp disease diagnostics.
Area of Science:
- Aquatic Animal Health
- Molecular Diagnostics
- Nanotechnology in Biosensing
Background:
- Infectious myonecrosis virus (IMNV) causes significant losses in Penaeus (Litopenaeus) vannamei aquaculture.
- Rapid and accurate detection of IMNV is crucial for disease management.
- Existing detection methods can be time-consuming or require specialized equipment.
Purpose of the Study:
- To develop a novel, rapid, and cost-effective colorimetric detection method for IMNV.
- To utilize gold nanoparticle (AuNP) probes combined with reverse transcription loop-mediated isothermal amplification (RT-LAMP).
- To establish a sensitive assay comparable to established molecular techniques.
Main Methods:
- A single-stranded DNA (ssDNA) probe labeled with gold nanoparticles (AuNPs) was designed.
- The assay relies on salt-induced AuNP aggregation for visual detection.
- IMNV-specific RT-LAMP products were amplified and detected using the AuNP probe system.
Main Results:
- The assay visually distinguished between IMNV presence (pink color) and absence (purple color) after salt addition.
- The detection limit was 10 copies of IMNV RNA, comparable to RT-PCR and LFD methods.
- The assay demonstrated 100-fold higher sensitivity than conventional RT-PCR methods.
Conclusions:
- The developed ssDNA-AuNP probe combined with RT-LAMP provides a rapid (approx. 50 min), sensitive, and visual detection method for IMNV.
- This assay is a promising low-cost tool for diagnosing IMNV in shrimp.
- The platform can be adapted for the detection of other shrimp pathogens.
