title: Development of a SYBR Green real-time RT-PCR assay for the detection of avian encephalomyelitis virus.
abstract: 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.
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China.
authors.forename: Qingtian
authors.initials: Q
authors.lastname: Liu
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China.
authors.forename: Zengqi
authors.initials: Z
authors.lastname: Yang
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China.
authors.forename: Huafang
authors.initials: H
authors.lastname: Hao
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China.
authors.forename: Shenli
authors.initials: S
authors.lastname: Cheng
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China.
authors.forename: Wentao
authors.initials: W
authors.lastname: Fan
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China.
authors.forename: Enqi
authors.initials: E
authors.lastname: Du
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China.
authors.forename: Sa
authors.initials: S
authors.lastname: Xiao
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China.
authors.forename: Xinglong
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A & F University, Yangling, Shaanxi Province, China. Electronic address: zhangshuxia316316@163.com.
authors.forename: Shuxia
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D002645:Chickens; D002681:China; D017931:DNA Primers / Q000235:genetics; D003959:Diamines; D001350:Encephalomyelitis Virus, Avian / Q000235:genetics / Q000302:isolation & purification*; D009930:Organic Chemicals / Q000378:metabolism*; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I
keywords: Avian encephalomyelitis virus; Real-time RT-PCR; SYBR Green