DNA probe and polymerase chain reaction procedure for the specific detection of Serpulina hyodysenteriae.
Serpulina (Treponema) hyodysenteriae, a Gram-negative anaerobic spirochete, is the causative agent of swine dysentery, a mucohaemorrhagic diarrheal disease in which lesions are confined to the large intestine of pigs. A DNA probe and polymerase chain reaction (PCR) amplification procedures which are specific, rapid , and sensitive for the detection of S.hyodysenteriae have been developed. Clone pF12 from a plasmid library of S.hyodysenteriae B204 genomic DNA was identified as a clone specific for S.hyodysenteriae but not for S.innocens by differential hybridization screening with S.hyodysenteriae and S.innocens genomic DNA probes. A DNA probe consisting of a 1.3 kb restriction fragment from pF12 was found to be highly specific for S. hyodysenteriae and detected 10(5) bacterial cells. A PCR procedure using primers derived from this fragment yielded a single product which was specifically generated for S.hyodysenteriae template DNA and not for other control cells DNA. PCR provided increased sensitivity with the direct detection of as few as 10 S.hyrodysenteriae cells. The PCR procedure could detect S.hyodysenteriae cells in seeded faecal matter. Moreover the PCR assay was able to detect most S. hyodysenteriae field isolates of serotypes 8 and 9. These tools have diagnostic application in veterinary microbiology.
Harel
J
J
GREMIP,Départment de Pathologie et Microbiologie, Faculté de Médecine Vétérinarire, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
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D000818:Animals; D001483:Base Sequence; D017401:Brachyspira hyodysenteriae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004403:Dysentery / Q000175:diagnosis / Q000382:microbiology / Q000821:virology; D005243:Feces / Q000382:microbiology; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012703:Serotyping; D013145:Spirochaetales Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D013552:Swine; D013553:Swine Diseases
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial