[DNA probe for detecting Yersinia pestis and serovariant I of Yersinia pseudotuberculosis by detecting specific DNA repeating sequences].
In order to construct a DNA probe for the plague pathogen detection, we have obtained the recombinant plasmid pRD100 carrying an EcoRI-flanked 140 bp fragment from the genetically silent region of Yersinia pestis species-specific plasmid pYP1. When used as a DNA probe for hybridization of DNA from various strains of 25 bacterial species, this DNA fragment was shown to have the complementary sequences in all investigated Yersinia pestis strains (200), including the plasmid pYP1 lacking ones, and in all the studied Yersinia pseudotuberculosis serotype I strains (80). The search for the probe target in these species has led us to conclusion that it is a specific repeated DNA sequence present in more copies in Yersinia pestis than in Yersinia pseudotuberculosis serotype I. The hybridization of these sequences with the radioactive probe and 24 hours autography makes possible the detection of 1.3 x 10(5) cells of Yersinia pestis and 3 x 10(6) cells of Yersinia pseudotuberculosis serotype I immobilized on the nitrocellulose membranes. Use of the probe for analysis of the nitrocellulose membrane fixed spleen smears from animals that died of experimental plague made possible the detection of Yersinia pestis cells within 48 h.
Podladchikova
O N
ON
Mishan'kin
B N
BN
Dikhanov
G G
GG
D001483:Base Sequence; D003001:Cloning, Molecular; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D016680:Genome, Bacterial; D008969:Molecular Sequence Data; D010957:Plasmids; D011995:Recombination, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D013045:Species Specificity; D015010:Yersinia pestis / Q000235:genetics*; D015011:Yersinia pseudotuberculosis / Q000235:genetics*
D015342:DNA Probes; D004269:DNA, Bacterial
DNK-zond dlia obnaruzheniia Yersinia pestis i I serovarianta Yersinia pseudotuberculosis metodom detektsii spetsificheskikh povtoriaiushchikhsiia posledovatel'nosteĭ DNK.