Development and evaluation of a one-step loop-mediated isothermal amplification for detection of spring viraemia of carp virus.
OBJECTIVE
Spring viraemia of carp virus (SVCV) is the causative agent of SVC disease. The main aim of our study was to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid, sensitive and effective detection of SVCV.


RESULTS
A set of four specific primers, two outer and two inner primers were designed based on the SVCV M gene for RT-LAMP assay. The sensitivity and specificity of RT-LAMP were determined and clinical test was performed under optimized amplification conditions (64 degrees C, 60 min). The results showed that the assay has a high specificity and the detection limit was 80 copies using 10-fold series dilutions of SVCV RNA, 10 times more sensitive than nest reverse transcription-polymerase chain reaction. In the detection of 472 fish samples, this assay showed excellent agreement with the standard virus isolation method (kappa = 0.807).


CONCLUSIONS
A sensitive and specific RT-LAMP assay was successfully developed to monitor and detect SVCV.


CONCLUSIONS
This work provides a robust method for evaluating the risk of SVCV. Given the advantages of LAMP in the detection of SVCV, this method can be applied to diagnose other viruses, which pose serious threats to the aquaculture industry.
Liu
Z
Z
College of Fisheries, Huazhong Agricultural University, Wuhan, China.
Teng
Y
Y
Xie
X
X
Li
H
H
Lv
J
J
Gao
L
L
Tian
F
F
Jiang
Y
Y
Chu
Z
Z
Xie
C
C
Liu
H
H
D000818:Animals; D017756:Aquaculture; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology; D005814:Genes, Viral; D021141:Nucleic Acid Amplification Techniques; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D018353:Rhabdoviridae Infections / Q000175:diagnosis* / Q000662:veterinary; D012680:Sensitivity and Specificity; D018116:Vesiculovirus / Q000235:genetics*; D014766:Viremia / Q000175:diagnosis / Q000821:virology