[cRNA probes (riboprobes) synthesized in vitro for detecting enterovirus by molecular hybridization].
Radioactively labelled RNA transcripts made in vitro of various fragments from cDNA clones of poliovirus type 1 and of hepatitis A virus under the control of bacteriophage T7 or SP6 promoters have been evaluated for diagnostic purposes. The RNA transcripts were 2 orders of magnitude more sensitive as hybridization probes than corresponding cDNA preparations labelled by the nick translation procedure. A combination of hybridization analysis and sequence comparison showed that some regions of the genome of a number of enteroviruses are highly conserved, while others show very little homology; the general order of conservation is: 5'-non-coding greater than 3'-terminal greater than central (2C) greater than VP3 greater than VP1. The 350 bases of the poliovirus VP1 region were highly specific for that virus, while the 450-bases of the 5'NC region showed extensive cross-reaction with other enteroviruses. However, these probes did not hybridize with HAV, which was detected only by HAV-specific riboprobes. The transcripts have been successfully applied as hybridization probes in diagnostic tests on supernatants of infected cell culture lysates and in clinical samples, mainly in stool extracts.
Kopecka
H
H
Virologie moléculaire, UA CNRS 545, Institut Pasteur, Paris, France.
Prévot
J
J
Girard
M
M
Fuchs
F
F
Aymard
M
M
D000818:Animals; D029822:Enterovirus B, Human / Q000032:analysis / Q000235:genetics; D004769:Enterovirus Infections / Q000175:diagnosis*; D005814:Genes, Viral; D006507:Hepatovirus / Q000032:analysis / Q000235:genetics; D006801:Humans; D009693:Nucleic Acid Hybridization; D017955:Poliovirus / Q000032:analysis / Q000235:genetics; D015347:RNA Probes; D012689:Sequence Homology, Nucleic Acid; D014709:Vero Cells
D015347:RNA Probes
Intérêt des sondes ARNc (ribosondes) synthétisés in vitro dans la détection des entérovirus par hybridation moléculaire.