Detection of avian reovirus RNA and comparison of a portion of genome segment S3 by polymerase chain reaction and restriction enzyme fragment length polymorphism.
A reverse transcription-polymerase chain reaction (RT-PCR) was established to amplify a 672-base pairs fragment on the segment S3 of avian reovirus (ARV). The amplified fragments were detected in nine strains of ARV as well as three tendon tissue specimens, indicating that the primer regions were well conserved. The RT-PCR was able to detect as low as 0.2 pg using an ethidium bromide stained gel. The detection limit could be enhanced further to 0.04 pg by hybridisation after southern transfer. The amplified DNA fragments from nine ARV strains and two tissue specimens showed different restriction enzyme cleavage patterns. Analysis of the data revealed that these 11 strains were classified into four groups. The results suggest that PCR followed by restriction enzyme analysis may provide a simple and rapid method for the characterisation of ARV isolates.
Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan.
L H
LH
Lee
J H
JH
Shien
H K
HK
Shieh
D000818:Animals; D001715:Bird Diseases / Q000821:virology*; D002642:Chick Embryo; D004591:Electrophoresis, Polyacrylamide Gel; D016679:Genome, Viral; D018165:Orthoreovirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012150:Polymorphism, Restriction Fragment Length; D011201:Poultry Diseases / Q000821:virology; D012367:RNA, Viral / Q000302:isolation & purification*; D012088:Reoviridae Infections / Q000662:veterinary* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012367:RNA, Viral