Genotyping of infectious bursal disease virus strains by restriction fragment length polymorphism analysis of the VP1, VP2, and VP3 genes.
SUMMARY. This study aimed to genotype infectious bursal disease virus (IBDV) isolates from the Minas Gerais state poultry industry. RNA was extracted from bursae obtained from field cases without passage or commercial vaccines. Genetic subtyping of IBDV isolates and vaccine strains was carried out by the reverse transcriptase-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP) analysis. A 588-bp fragment in the VP1 gene, an 847-bp fragment in the VP2 gene, and a 320-bp fragment in the VP3 gene were amplified by PCR and digested with restriction enzymes PstI and ScaI (VP1); BamHI, BstEII, and PstI (VP2); and NcoI, ScaI, and XbaI (VP3). Our work shows that complementing the clinical history of the outbreaks with RT-PCR followed by RFLP analysis using PstI for VP1, BamHI for VP2, and XbaI for VP3 allowed an accurate classification of a causative agent as a very virulent IBDV.
Gomes
A D
AD
Departamento de Microbiologia, Laboratório de Virologia Comparada Instituto de Ciências Biológicas, PO Box 486, Belo Horizonte, Minas Gerais, Brazil.
Abreu
J T
JT
Redondo
R A F
RA
Martins
N R S
NR
Resende
J S
JS
Resende
M
M
D000818:Animals; D001483:Base Sequence; D018175:Birnaviridae Infections / Q000662:veterinary / Q000821:virology; D001938:Brazil; D002645:Chickens / Q000821:virology*; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D014644:Genetic Variation; D005838:Genotype; D007243:Infectious bursal disease virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D010802:Phylogeny; D012150:Polymorphism, Restriction Fragment Length; D011201:Poultry Diseases / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015678:Viral Structural Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000235:genetics
D004279:DNA, Viral; C107669:VP1 protein, infectious bursal disease virus; C082513:VP2 protein, infectious bursal disease virus; C478244:VP3 protein, infectious bursal disease virus; D015678:Viral Structural Proteins; D014765:Viral Vaccines