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Colorimetric detection of scale drop disease virus in Asian sea bass using loop-mediated isothermal amplification with xylenol orange.

Authors :
Dangtip, Sirintip
Kampeera, Jantana
Suvannakad, Rapheephat
Khumwan, Pakapreud
Jaroenram, Wansadaj
Sonthi, Molruedee
Senapin, Saengchan
Kiatpathomchai, Wansika
Source :
Aquaculture. Aug2019, Vol. 510, p386-391. 6p.
Publication Year :
2019

Abstract

Scale drop disease virus (SDDV) is an emerging pathogen that causes multiple outbreaks in farmed Asian sea bass across many regions in Southeast Asia and results in severe economic losses to the exports of fish. With no current standard vaccination protocol against this virus, routine testing that probes for SDDV in Asian sea bass culture can help farmers prevent a large-scale epidemic from occurring. Fish samples can be sent to accredited laboratories for SDDV test that is performed via polymerase chain reaction (PCR) and related methods, such as qPCR. Albeit its high sensitivity, PCR requires skilled labors and equipment only available in centralized laboratory settings, and therefore, not appropriate for the timely determination of SDDV. Herein, we report the development of a colorimetric assay based on loop-mediated isothermal amplification (LAMP) with a pH-sensitive xylenol orange (XO) reporter. With a rapid total DNA extraction method developed in this research, we were able to demonstrate that the sensitivity of the naked-eye detection was 100 picograms of total DNA extracted from the SDDV-infected Asian sea bass samples. In addition to its affordability, our LAMP-XO circumvents the technical challenges faced by PCR and allows the SDDV detection to be conveniently performed and analyzed at the potential site of an outbreak. • A colorimetric LAMP assay with pH-sensitive xylenol orange (XO) reporter was developed to detect SDDV in Asian sea bass. • The limit of naked-eye detection of LAMP-XO was 100 pg of total DNA extracted from the SDDV-infected Asian sea bass samples. • The technique was validated by field SDDV-infected and healthy Asian sea bass samples. • The technique is compatible with a rapid DNA extraction protocol using G3T buffer. • The LAMP-XO assay has a potential use in an active surveillance of L. calcarifer. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
00448486
Volume :
510
Database :
Academic Search Index
Journal :
Aquaculture
Publication Type :
Academic Journal
Accession number :
141581102
Full Text :
https://doi.org/10.1016/j.aquaculture.2019.05.071