Development of a reverse transcription loop-mediated isothermal amplification assay for visual detection of avian reovirus

被引：20

作者：

Xie, Zhixun ^{[1
]}

Peng, Yi ^{[1
]}

Luo, Sisi ^{[1
]}

Wang, Ying ^{[1
]}

Liu, Jiabo ^{[1
]}

Pang, Yaoshan ^{[1
]}

Deng, Xianwen ^{[1
]}

Xie, Zhiqin ^{[1
]}

Xie, Liji ^{[1
]}

Fan, Qing ^{[1
]}

Teng, Liqiong ^{[1
]}

Wang, Xiuqing ^{[2
]}

机构：

[1] Guangxi Vet Res Inst, Guangxi Key Lab Anim Vaccines & New Technol, Dept Biotechnol, Nanning 530001, Guangxi, Peoples R China

[2] S Dakota State Univ, Dept Biol & Microbiol, Brookings, SD 57007 USA

来源：

AVIAN PATHOLOGY | 2012年 / 41卷 / 03期

基金：

中国国家自然科学基金;

关键词：

RAPID DETECTION; VIRUS; IDENTIFICATION; CHICKENS; PRODUCTS; GENE; LAMP; PCR;

D O I：

10.1080/03079457.2012.686104

中图分类号：

S85 [动物医学（兽医学）];

学科分类号：

0906 ;

摘要：

Avian reovirus (ARV) is an important pathogen of poultry and causes significant economic losses to the poultry industry. To develop a rapid and sensitive method for the surveillance of ARV, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was established using a set of six primers specific to the S1 gene segment of ARV. The established assay was performed at 62 degrees C for 60 min in a thermal block, and the result was visualized directly under daylight or ultraviolet light. The detection limit of the RT-LAMP assay was 10 fg total RNA, which was 100-fold higher than that of reverse transcriptase polymerase chain reactions. The specificity of the assay was supported by the lack of cross-reaction with other avian pathogens. Furthermore, viral RNAs of field isolates were successfully detected by the assay. Overall, the newly established RT-LAMP assay is simple, rapid, sensitive, specific, and can visually detect ARV without the use of any specialized equipment.

引用

页码：311 / 316

页数：6

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