Visual Detection of West Nile Virus Using Reverse Transcription Loop-Mediated Isothermal Amplification Combined with a Vertical Flow Visualization Strip

被引：23

作者：

Cao, Zengguo ^{[1
]}

Wang, Hualei ^{[1
,2
]}

Wang, Lina ^{[1
,3
]}

Li, Ling ^{[1
,4
]}

Jin, Hongli ^{[1
,5
]}

Xu, Changping ^{[1
,6
]}

Feng, Na ^{[1
,2
]}

Wang, Jianzhong ^{[1
]}

Li, Qian ^{[1
]}

Zhao, Yongkun ^{[1
]}

Wang, Tiecheng ^{[1
]}

Gao, Yuwei ^{[1
,2
]}

Lu, Yiyu ^{[1
,6
]}

Yang, Songtao ^{[1
,2
]}

Xia, Xianzhu ^{[1
,2
]}

机构：

[1] Acad Mil Med Sci, Key Lab Jilin Prov Zoonosis Prevent & Control, Inst Mil Vet, Changchun, Peoples R China

[2] Jiangsu Coinnovat Ctr Prevent & Control Important, Yangzhou, Jiangsu, Peoples R China

[3] Jilin Agr Univ, Anim Sci & Technol Coll, Changchun 130122, Peoples R China

[4] Jilin Univ, Coll Vet Med, Changchun 130023, Peoples R China

[5] Changchun SR Biol Technol Co Ltd, Changchun, Peoples R China

[6] Zhejiang Prov Ctr Dis Control & Prevent, Key Lab Emergency Detect Publ Hlth Zhejiang Prov, Hangzhou, Zhejiang, Peoples R China

来源：

FRONTIERS IN MICROBIOLOGY | 2016年 / 7卷

关键词：

West Nile virus; visual detection; reverse transcription loop-mediated isothermal amplification; visualization strip; RT-LAMP-VF; TIME RT-PCR; ENCEPHALITIS-VIRUS; LINEAGES; INFECTION; FEATURES; ASSAY;

D O I：

10.3389/fmicb.2016.00554

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

West Nile virus (WNV) causes a severe zoonosis, which can lead to a large number of casualties and considerable economic losses. A rapid and accurate identification method for WNV for use in field laboratories is urgently needed. Here, a method utilizing reverse transcription loop-mediated isothermal amplification combined with a vertical flow visualization strip (RT-LAMP-VF) was developed to detect the envelope (E) gene of WNV. The RT-LAMP-VF assay could detect 10(2) copies/mu l of an WNV RNA standard using a 40 min amplification reaction followed by a 2 min incubation of the amplification product on the visualization strip, and no cross-reaction with other closely related members of the Flavivirus genus was observed. The assay was further evaluated using cells and mouse brain tissues infected with a recombinant rabies virus expressing the E protein of WNV. The assay produced sensitivities of 10(1.5) TCID50/ml and 10(1.33) TCID50/ml for detection of the recombinant virus in the cells and brain tissues, respectively. Overall, the RT-LAMP-VF assay developed in this study is rapid, simple and effective, and it is therefore suitable for clinical application in the field.

引用

页数：9

共 34 条

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