Detection of genetic variations in serotype I isolates of infectious bursal disease virus using polymerase chain reaction and restriction endonuclease analysis.
Reverse transcription with polymerase chain reaction (PCR) followed by restriction endonuclease analysis detected genetic variations among serotype I isolates of infectious bursal disease virus (IBDV). Using a set of synthetic primers derived from the large genome segment of APHIS-IBDV, the hypervariable region (AccI-SpeI fragment) located in the VP2 gene was amplified. With all strains, a cDNA fragment of approximately 643 bp was amplified, indicating that there were no apparent deletions or insertions in this region among isolates. Fragments amplified from 9 isolates were digested with 14 restriction enzymes. Restriction fragment profiles generated by restriction enzymes NaeI, StuI, TaqI, and SacI, showed genetic variations among isolates. This study provided a simple and sensitive method for detection of genetic variations among isolates that are closely related serologically and could not be differentiated using current serologic methods.
Liu
H J
HJ
Department of Poultry Science, Auburn University, AL 36849-5416.
Giambrone
J J
JJ
Dormitorio
T
T
D000818:Animals; D001483:Base Sequence; D018175:Birnaviridae Infections / Q000662:veterinary / Q000821:virology; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D004262:DNA Restriction Enzymes; D018076:DNA, Complementary / Q000235:genetics; D014644:Genetic Variation; D007243:Infectious bursal disease virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011201:Poultry Diseases / Q000821:virology; D012703:Serotyping; D014773:Virology / Q000379:methods
D017931:DNA Primers; D018076:DNA, Complementary; D004262:DNA Restriction Enzymes