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Reverse transcription loop-mediated isothermal amplification for rapid detection of Japanese encephalitis virus in swine and mosquitoes.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Liu, Hao Liu, Zhen-Jiang Jing, Jie Ren, Jing-Qiang Liu, Yan-Yu Guo, Huan-Huan Fan, Min Lu, Hui-Jun Jin, Ning-Yi
Description	Author Affiliation: Liu H ( College of Animal Science and Veterinary Medicine, Jilin University, Changchun, PR China.)
Abstract	Japanese encephalitis (JE) can infect many agriculturally important animals and humans, and has a high incidence in Asia. One of the natural hosts of the mosquito-borne JE virus (JEV) is domestic pigs, which act as amplifier hosts. Porcine infection results in fatal encephalitis, abortion, and stillbirth in pregnant sows, and hypospermia in boars. In this study, a rapid JEV detection method for swine and mosquitoes was developed based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) targeting the nucleocapsid (E) genes of JEV genotype I (lineage K94PO5), and genotype III (lineage SA14-14-2). Fifty-six swine blood samples and 20,000 mosquitoes were used to evaluate the method, compared to conventional RT-polymerase chain reaction (PCR) and real-time RT-PCR. RT-LAMP had detection limits of 2.57 and 2.34 copies/µL for JEV I and III, respectively. Assay sensitivity was similar to real-time RT-PCR, but was 10-fold higher than conventional RT-PCR. Assay specificity was high, showing no cross-reactivity to other flaviviruses. The results of virus isolation and identification of swine blood samples and mosquito samples were fully consistent with RT-LAMP. Finally, the JEV RT-LAMP assay was simpler and less time consuming than conventional RT-PCR or real-time RT-PCR, since the amplification step could be completed in a single tube within 50 min at 63°C. In conclusion, the newly-developed RT-LAMP assay is an accurate and convenient method for rapid and sensitive diagnosis of JEV in swine and mosquitoes, and may prove to be a practical molecular tool for surveillance and epidemiological investigations.
File Format	HTM / HTML
ISSN	15303667
e-ISSN	15577759
DOI	10.1089/vbz.2012.0991
Journal	Vector-Borne and Zoonotic Diseases
Issue Number	12
Volume Number	12
Language	English
Publisher	Mary Ann Liebert, Inc.
Publisher Date	2012-12-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Communicable Diseases Discipline Epidemiology Culicidae Virology Encephalitis Virus, Japanese Isolation & Purification Encephalitis, Japanese Veterinary Insect Vectors Nucleocapsid Proteins Genetics Swine Diseases Diagnosis Animals Dna Primers Disease Reservoirs Epidemiology Limit Of Detection Pregnancy Rna, Viral Blood Reference Standards Reverse Transcriptase Polymerase Chain Reaction Reverse Transcription Sensitivity And Specificity Swine Time Factors Zoonoses Comparative Study Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Infectious Diseases Virology Microbiology

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