Detection of enteroviruses using subgenomic probes of Coxsackie virus B4 by hybridization. 1988

N K Chatterjee, and M Kaehler, and R Deibel
Laboratories for Virology, Wadsworth Center for Laboratories and Research, Albany, NY 12201.

The objective of this research was to develop group- and type-specific probes for the detection of enteroviruses. Coxsackie virus B4 RNA was cloned, and a series of subgenomic clones were generated. Six of these clones, containing sequences from the 3' end or the 5' end of the genome, were tested for their ability to detect these viruses in a small number of infected cells employing nucleic acid hybridization technique and total cytoplasmic RNA from a panel of 11 serotypes of enteroviruses. The RNA from cells infected with Coxsackie B viruses gave characteristic and positive hybridization signals. Coxsackie B-specific probes and a control Echo 9 probe detected Coxsackie A9 and Echo 3 weakly. As little as 0.5 microgram of the RNA--which contained 10-20 ng of poly(A)-containing, virus-specific, hybridizable RNA--was sufficient to successfully conduct the assay, suggesting high sensitivity of these probes. Probes that are 3' end-specific appear to be group specific, while those that are 5' end-specific appear to be type specific among the serotypes tested.

UI MeSH Term Description Entries
D009693 Nucleic Acid Hybridization Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503) Genomic Hybridization,Acid Hybridization, Nucleic,Acid Hybridizations, Nucleic,Genomic Hybridizations,Hybridization, Genomic,Hybridization, Nucleic Acid,Hybridizations, Genomic,Hybridizations, Nucleic Acid,Nucleic Acid Hybridizations
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D004770 Enterovirus A genus of the family PICORNAVIRIDAE whose members preferentially inhabit the intestinal tract of a variety of hosts. The genus contains many species. Newly described members of human enteroviruses are assigned continuous numbers with the species designated "human enterovirus". Coxsackie Viruses,Coxsackieviruses
D005814 Genes, Viral The functional hereditary units of VIRUSES. Viral Genes,Gene, Viral,Viral Gene
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D015342 DNA Probes Species- or subspecies-specific DNA (including COMPLEMENTARY DNA; conserved genes, whole chromosomes, or whole genomes) used in hybridization studies in order to identify microorganisms, to measure DNA-DNA homologies, to group subspecies, etc. The DNA probe hybridizes with a specific mRNA, if present. Conventional techniques used for testing for the hybridization product include dot blot assays, Southern blot assays, and DNA:RNA hybrid-specific antibody tests. Conventional labels for the DNA probe include the radioisotope labels 32P and 125I and the chemical label biotin. The use of DNA probes provides a specific, sensitive, rapid, and inexpensive replacement for cell culture techniques for diagnosing infections. Chromosomal Probes,DNA Hybridization Probe,DNA Probe,Gene Probes, DNA,Conserved Gene Probes,DNA Hybridization Probes,Whole Chromosomal Probes,Whole Genomic DNA Probes,Chromosomal Probes, Whole,DNA Gene Probes,Gene Probes, Conserved,Hybridization Probe, DNA,Hybridization Probes, DNA,Probe, DNA,Probe, DNA Hybridization,Probes, Chromosomal,Probes, Conserved Gene,Probes, DNA,Probes, DNA Gene,Probes, DNA Hybridization,Probes, Whole Chromosomal
D017955 Poliovirus A species of ENTEROVIRUS which is the causal agent of POLIOMYELITIS in humans. Three serotypes (strains) exist. Transmission is by the fecal-oral route, pharyngeal secretions, or mechanical vector (flies). Vaccines with both inactivated and live attenuated virus have proven effective in immunizing against the infection. Brunhilde Virus,Human poliovirus 1,Human poliovirus 2,Human poliovirus 3,Lansing Virus,Leon Virus,Poliovirus Type 1,Poliovirus Type 2,Poliovirus Type 3,Polioviruses
D029822 Enterovirus B, Human A species of ENTEROVIRUS infecting humans and containing 36 serotypes. It is comprised of all the echoviruses and a few coxsackieviruses, including all of those previously named coxsackievirus B. Bornholm Disease Virus,Coxsackievirus A9, Human,Echoviruses,Pleurodynia Virus,Coxsackie A9 Virus,Coxsackie B Viruses,Coxsackieviruses B,Echo Viruses,Human Coxsackievirus A9,Human Enterovirus B,Swine Vesicular Disease Virus,A9 Viruses, Coxsackie,Coxsackie A9 Viruses,Coxsackie B Virus,Disease Virus, Bornholm,Echo Virus,Echovirus,Pleurodynia Viruses,Virus, Bornholm Disease,Virus, Coxsackie A9,Virus, Coxsackie B,Viruses, Coxsackie A9

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