Identification of human rotavirus serotype by hybridization to polymerase chain reaction-generated probes derived from a hyperdivergent region of the gene encoding outer capsid protein VP7.
We have synthesized 32P-labeled hybridization probes from a hyperdivergent region (nucleotides 51 to 392) of the rotavirus gene encoding the VP7 glycoprotein by using the polymerase chain reaction method. Both RNA (after an initial reverse transcription step) and cloned cDNA from human rotavirus serotypes 1 through 4 could be used as templates to amplify this region. High-stringency hybridization of each of the four probes to rotavirus RNAs dotted on nylon membranes allowed the specific detection of corresponding sequences and thus permitted identification of the serotype of the strains dotted. The procedure was useful when applied to rotaviruses isolated from field studies.
Flores
J
J
Laboratory of Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Sears
J
J
Schael
I P
IP
White
L
L
Garcia
D
D
Lanata
C
C
Kapikian
A Z
AZ
D000956:Antigens, Viral / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D004279:DNA, Viral / Q000235:genetics; D005243:Feces / Q000382:microbiology; D005814:Genes, Viral; D006801:Humans; D007231:Infant, Newborn; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D010761:Phosphorus Radioisotopes; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000235:genetics; D012401:Rotavirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D012703:Serotyping; D015678:Viral Structural Proteins / Q000235:genetics*; D014765:Viral Vaccines
D000956:Antigens, Viral; D036022:Capsid Proteins; D004279:DNA, Viral; D015345:Oligonucleotide Probes; D010761:Phosphorus Radioisotopes; D012367:RNA, Viral; C050779:VP7 protein, Rotavirus; D015678:Viral Structural Proteins; D014765:Viral Vaccines