A simple and rapid method for detection of Goose Parvovirus in the field by loop-mediated isothermal amplification.
BACKGROUND
Goose parvovirus (GPV) is a Dependovirus associated with latent infection and mortality in geese. Currently, in a worldwide scale, GPV severely affects geese production. The objective of this study is to develop a loop-mediated isothermal amplification (LAMP) method for the sensitive, rapid, and inexpensive detection of GPV in the field.


RESULTS
A set of six specific primers was designed by targeting the GPV VP3 DNA. With Bst DNA polymerase large fragment, the target DNA could be amplified at 65 degrees C as early as 20 min of incubation in a simple water bath. A positive reaction was identified through the detection of the LAMP product by color change visible to the naked eye. The detection limit of the assay was 28 copies/microl of plasmid pVP3, and with equal sensitivity and specificity to fluorescent quantitative real-time PCR (FQ-PCR).


CONCLUSIONS
The high sensitivity, specificity, and simplicity, as well as the high throughput, make this method suitable for specific detection of GPV infection in both field conditions and laboratory settings. The utilization of complicated equipment and conduct of technical training on the GPV LAMP were not necessary.
Yang
JinLong
J
Chongqing Academy of Animal Science, Chongqing 402460, Chongqing, China.
Yang
Rui
R
Cheng
AnChun
A
Wang
MingShu
M
Fu
LiZhi
L
Yang
SongQuan
S
Zhang
SuHui
S
Yang
Liu
L
Xu
ZhiYong
Z
D000818:Animals; D052160:Benzothiazoles; D017931:DNA Primers / Q000235:genetics; D000229:Dependovirus / Q000235:genetics / Q000302:isolation & purification*; D003959:Diamines; D005777:Geese; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009930:Organic Chemicals / Q000378:metabolism; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D011201:Poultry Diseases / Q000821:virology*; D011804:Quinolines; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013696:Temperature; D015678:Viral Structural Proteins / Q000235:genetics
D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D015678:Viral Structural Proteins; C098022:SYBR Green I