Development of a SYBR Green real-time RT-PCR assay for the detection of avian encephalomyelitis virus
| Autor: | Sa Xiao, Wentao Fan, Shuxia Zhang, Shenli Cheng, Enqi Du, Xinglong Wang, Qingtian Liu, Huafang Hao, Zengqi Yang |
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| Rok vydání: | 2014 |
| Předmět: |
China
Avian encephalomyelitis animal structures Diamines Biology Real-Time Polymerase Chain Reaction Sensitivity and Specificity Rapid detection Virus chemistry.chemical_compound Virology Animals Benzothiazoles Organic Chemicals Poultry Diseases DNA Primers Picornaviridae Infections Staining and Labeling Reverse Transcriptase Polymerase Chain Reaction RNA Vp1 gene Molecular biology Real-time polymerase chain reaction Encephalomyelitis Virus Avian chemistry Specific primers embryonic structures Quinolines Chickens DNA |
| Zdroj: | Journal of Virological Methods. 206:46-50 |
| ISSN: | 0166-0934 |
| Popis: | Avian encephalomyelitis virus (AEV) causes epidemic diseases in poultry worldwide. A SYBR Green real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay was developed for the rapid detection and quantitation of AEV in this study. A pair of specific primers was designed in the highly conserved VP1 gene of this virus. When comparing this assay with conventional RT-PCR, the rRT-PCR assay was 100 times more sensitive and could detect levels as low as 10 standard DNA copies of the AEV SX strain. The specificity of this technique was evaluated in five other avian pathogens. The AEV RNA was detected as early as three days post-infection in chicken embryos. All 18 clinical chicken brains collected from an AEV outbreak in Northwestern China were detected to be positive (100%) using the rRT-PCR assay. However, only 5 of the 18 samples were positive (28%) using the conventional RT-PCR. The results were confirmed by virus isolation in chicken embryos. This high sensitivity, specificity, and simplicity of the SYBR Green rRT-PCR approach can be a more effective method than the conventional one for AEV diagnosis and surveillance. |
| Databáze: | OpenAIRE |
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