Restriction endonucleases whose sites are predictable from the amino acid sequence offer an improved strategy for typing bovine rotaviruses.
Variation in the third base of a codon hampers genotypic characterization, particularly of RNA viruses. Some restriction endonucleases, however, have a recognition site with a variable base at the third position and will always cleave when a certain amino acid pair occurs (such as glycine-proline for Sau96I and glutamic or aspartic acid followed by serine usually for HinfI). We developed a restriction fragment length polymorphism (RFLP) procedure based on these enzymes for P-typing bovine group A rotaviruses (BRV). Employing this procedure 20 BRV local strains, isolated in tissue culture as well as the original faecal sample, could be typed in one of three patterns. More variability was observed when restriction endonucleases were employed whose cleavage sites cannot be predicted from the amino acid sequence (TaqI and Tsp509I). These RFLP results agreed with the PCR-VP4 typing assay, neutralization tests, and nucleotide sequence analysis. RFLP with Sau96I and HinfI provided quick and objective P-typing of strains and could detect multiple genotypes in the same sample.
Hussein
H A
HA
Section de virologie, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada.
Frost
E H
EH
Deslandes
S
S
Talbot
B
B
Elazhary
Y
Y
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000821:virology; D017931:DNA Primers; D004262:DNA Restriction Enzymes / Q000378:metabolism; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D005838:Genotype; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D011792:Quebec; D012367:RNA, Viral / Q000032:analysis; D015183:Restriction Mapping; D012401:Rotavirus / Q000145:classification* / Q000235:genetics; D012400:Rotavirus Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology
D017931:DNA Primers; D018076:DNA, Complementary; D012367:RNA, Viral; D004262:DNA Restriction Enzymes