Rapid and sensitive detection of Singapore grouper iridovirus by loop-mediated isothermal amplification.
OBJECTIVE
The aim of this paper was to develop a loop-mediated isothermal amplification (LAMP) method for rapid, sensitive and inexpensive detection of Singapore grouper iridovirus (SGIV) in grouper (GP), Epinephelus sp.


RESULTS
A set of six specific primers was designed by targeting the SGIV ORF-014L. With Bst DNA polymerase large fragment, the target DNA can be amplified as early as 20 min at 65 degrees C in a simple water bath. The detection limit is about 0.02 fg (equivalent to 6.3 copies) of plasmid ORF-014L. LAMP products could be judged with three different methods. There were no cross-reactions with seven other aquatic animal viruses indicating high specificity of the LAMP. The LAMP method was applied to detect SGIV in virus-infected GP cells and GP tissues effectively.


CONCLUSIONS
The LAMP described in this study is a cheap, sensitive, specific and rapid protocol for the detection of SGIV in cells and in GP tissues.


CONCLUSIONS
The developed LAMP method can be simply applied both in field condition and in laboratory operation for specific detection of SGIV infection.
Mao
X L
XL
State Key Laboratory of Biocontrol, College of Life Science, Sun Yat-sen University, Guangzhou, PR China.
Zhou
S
S
Xu
D
D
Gong
J
J
Cui
H C
HC
Qin
Q W
QW
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis*; D005399:Fishes / Q000821:virology*; D005516:Food Microbiology; D017994:Iridovirus / Q000235:genetics*; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods; D014773:Virology / Q000379:methods
D017931:DNA Primers; D004279:DNA, Viral