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PMID: 2164610 Published · ppublish English Journal Article

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.

Journal of virology ·Vol. 64 ·No. 8 ·1990-08-00 ·Pages 4021-4

Flores J, Sears J, Schael IP, White L, Garcia D, Lanata C, Kapikian AZ

Abstract

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.

MeSH Terms
Antigens, Viral/genetics Base Sequence Blotting, Northern Capsid/genetics Capsid Proteins DNA, Viral/genetics Feces/microbiology Genes, Viral Humans Infant, Newborn Molecular Sequence Data Nucleic Acid Hybridization Oligonucleotide Probes Phosphorus Radioisotopes Polymerase Chain Reaction/methods RNA, Viral/genetics Rotavirus/classification,genetics,isolation & purification Serotyping Viral Structural Proteins/genetics Viral Vaccines
Chemicals
Antigens, Viral Capsid Proteins DNA, Viral Oligonucleotide Probes Phosphorus Radioisotopes RNA, Viral VP7 protein, Rotavirus Viral Structural Proteins Viral Vaccines
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Flores J
Laboratory of Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Sears J
Schael I P
White L
Garcia D
Lanata C
Kapikian A Z
References (12)
12 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
Published
1990-08-00
Pages
4021-4
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC249704
Subset
IM
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