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Updated: Jun 22, 2026

Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay for the Specific and Rapid Detection of Tilapia Lake Virus

Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay for the Specific and Rapid Detection of Tilapia Lake Virus

Published on: May 18, 2020

Rapid, sensitive and simple detection method for koi herpesvirus using loop-mediated isothermal amplification.

Manabu Yoshino¹, Hajime Watari, Tadashi Kojima

¹Biochemical Research Laboratory, Eiken Chemical Co., Ltd, Tochigi 329-0114, Japan. manabu_yoshino@eiken.co.jp

Microbiology and Immunology

|July 1, 2009

Summary

New Loop-mediated isothermal amplification (LAMP) methods offer sensitive and specific detection of koi herpesvirus (KHV, CyHV-3). The SphI-5 LAMP assay is recommended for routine diagnostics due to its rapid results.

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Area of Science:

Aquaculture
Veterinary Virology
Molecular Diagnostics

Background:

Koi herpesvirus (KHV, CyHV-3) causes significant mortality in koi and common carp.
Accurate and rapid diagnostic tools are crucial for managing KHV outbreaks in aquaculture.

Purpose of the Study:

To develop and evaluate Loop-mediated isothermal amplification (LAMP) assays for KHV detection.
To compare the sensitivity and specificity of LAMP assays against established PCR methods.
To assess the suitability of LAMP for routine KHV diagnostics.

Main Methods:

Two primer sets (9/5 LAMP and SphI-5 LAMP) were designed targeting conserved KHV sequences.
Isothermal amplification was performed at 65°C for 60 minutes, monitoring turbidity changes.
LAMP assays were tested for detection limits and cross-reactivity with other fish pathogens.
Performance was compared with a modified SphI-5 PCR assay.

Main Results:

Both LAMP assays achieved a detection limit of six copies, matching the modified SphI-5 PCR.
No cross-reactivity was observed with other fish viruses and bacteria.
The SphI-5 LAMP assay demonstrated a faster reaction velocity compared to the 9/5 LAMP assay.
LAMP assays were effective using crude extracts from fish organs.

Conclusions:

LAMP offers a sensitive, specific, rapid, and simple method for KHV detection.
The SphI-5 LAMP assay is a superior option for routine KHV diagnostics.
LAMP represents a valuable tool for diagnosing KHV infections in fish populations.