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Development of the Visual Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD) Method for Rapid Detection of Vibrio harveyi in Aquatic Animals
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Development of the Visual Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD) Method for Rapid Detection of Vibrio harveyi in Aquatic Animals
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Development of the Visual Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD) Method for Rapid Detection of Vibrio harveyi in Aquatic Animals
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Journal Article
Development of the Visual Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick (LAMP-LFD) Method for Rapid Detection of Vibrio harveyi in Aquatic Animals
2024
Overview
Vibrio harveyi
is a normal flora present in seawater, but in recent years, with large density aquaculture,
V. harveyi
has become one of the causative organisms in fish and shrimp. In this study,
V. harveyi
VieA
gene was used as the target to design primers and establish the loop-mediated isothermal amplification
(
LAMP) combined with visual lateral flow test strip (LFD). The optimal reaction conditions were optimized as 60°C, 45 min, 1.2 mM Mg
2+
, 0.64 mM dNTPs, 0.25 mM betaine, and 16:1 ratio of internal/external primers. The results showed that the established LAMP assay was able to specifically detect
V. harveyi
with a sensitivity of 3.4 × 10
–5
ng/µL. Clinical applicability analysis revealed that only
V. harveyi
was detected in the samples for pearl gentian grouper and whiteleg shrimp artificially infected by
Vibrio
. Therefore, we established a visual, reliable, rapid and sensitive LAMP-LFD method for detecting
V. harveyi
in aquatic animals.
Publisher
Pleiades Publishing,Springer Nature B.V
Subject
