Development of reverse-transcription loop-mediated isothermal amplification for the detection of infectious bursal disease virus.
To establish a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of infectious bursal disease virus (IBDV), four primers specific to six regions of the VP3 gene were designed; the VP3 region was selected because it is a conserved part of the IBDV genome. After amplification in an isothermal water bath for 70 min, samples containing IBDV generated the expected ladder-like products while other viruses generated no product. The sensitivity and specificity of the RT-LAMP assay were evaluated by comparison with reverse-transcription polymerase chain reaction (RT-PCR) and virus isolation. The assay was significantly more sensitive than normal gel-based RT-PCR. Because it is specific and simple, the RT-LAMP assay can be widely applied in clinical laboratories for rapid detection of IBVD.
Xu
Jiangtao
J
College of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
Zhang
Zhenmei
Z
Yin
Yanbo
Y
Cui
Shangjin
S
Xu
Shouzhen
S
Guo
Yanyan
Y
Li
Jida
J
Wang
Jianlin
J
Liu
Xingcai
X
Han
Limin
L
D000818:Animals; D018175:Birnaviridae Infections / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D002642:Chick Embryo; D002645:Chickens / Q000821:virology; D017931:DNA Primers; D007243:Infectious bursal disease virus / Q000235:genetics / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011201:Poultry Diseases / Q000175:diagnosis / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D015678:Viral Structural Proteins / Q000235:genetics
D017931:DNA Primers; C478244:VP3 protein, infectious bursal disease virus; D015678:Viral Structural Proteins