Reverse transcription loop-mediated isothermal amplification for the rapid detection of infectious bronchitis virus in infected chicken tissues.
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay targeting the nucleocapsid phosphoprotein gene of infectious bronchitis virus (IBV) was developed. The detection limits for the IBV RT-LAMP assay were 10(1) 50% egg infection dose (EID(50)) per 50 microl of titrated viruses and no cross-reaction of IBV RT-LAMP was found when tested with other viruses including Newcastle disease virus (NDV), avian reovirus (ARV), and infectious laryngotrachietis virus (ILTV) due to their mismatch with IBV RT-LAMP primers. A total of 187 clinical tissues samples (88 blood, 62 kidney and 37 lung) were evaluated and compared to conventional RT-PCR. The sensitivity of RT-LAMP and RT-PCR assays for detecting IBV RNA in clinical specimens was 99.5% and 98.4%, respectively. These findings showed that the RT-LAMP assay has potential usefulness for rapid and sensitive diagnosis in outbreak of IBV.
Chen
Hao-tai
HT
Key Laboratory of Animal Virology of Ministry of Agriculture, Key Laboratory of Veterinary Public Health of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
Zhang
Jie
J
Ma
Yan-ping
YP
Ma
Li-Na
LN
Ding
Yao-zhong
YZ
Liu
Xiang-tao
XT
Cai
Xue-peng
XP
Ma
Li-qing
LQ
Zhang
Yong-guang
YG
Liu
Yong-sheng
YS
D000818:Animals; D001483:Base Sequence; D002645:Chickens / Q000821:virology*; D018352:Coronavirus Infections / Q000662:veterinary* / Q000821:virology; D017931:DNA Primers; D004587:Electrophoresis, Agar Gel; D001351:Infectious bronchitis virus / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods*; D011201:Poultry Diseases / Q000821:virology*; D048348:Reverse Transcription / Q000235:genetics*; D012680:Sensitivity and Specificity; D013696:Temperature
D017931:DNA Primers