Reverse transcription loop-mediated isothermal amplification for the rapid detection of infectious bronchitis virus in infected chicken tissues

被引:32
作者
Chen, Hao-tai
Zhang, Jie
Ma, Yan-ping
Ma, Li-na
Ding, Yao-zhong
Liu, Xiang-tao
Cai, Xue-peng
Ma, Li-qing [2 ]
Zhang, Yong-guang
Liu, Yong-sheng [1 ]
机构
[1] Chinese Acad Agr Sci, Lanzhou Vet Res Inst, State Key Lab Vet Etiol Biol,Key Lab Anim Virol, Key Lab Vet Publ Hlth,Minist Agr,Dept Virol, Lanzhou 730046, Gansu, Peoples R China
[2] Qinghai Univ, Qinghai Acad Anim Sci & Vet Med, Xining 810003, Peoples R China
基金
中国国家自然科学基金;
关键词
Infectious bronchitis virus (IBV); Detection; Reverse transcription loop-mediated isothermal amplification (RT-LAMP); POLYMERASE-CHAIN-REACTION; SEROTYPE IDENTIFICATION; S-1; GENE; RT-PCR; ANTIBODIES; ASSAY;
D O I
10.1016/j.mcp.2009.10.001
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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 (EID50) per 50 mu l 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. (C) 2009 Elsevier Ltd. All rights reserved.
引用
收藏
页码:104 / 106
页数:3
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