Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
1994-10-6
pubmed:abstractText
A PCR assay for the detection of Serpulina hyodysenteriae in diagnostic specimens was developed on the basis of sequence analysis of a recombinant clone designated pRED3C6. Clone pRED3C6, which contained a 2.3-kb DNA fragment unique to S. hyodysenteriae, was identified by screening a plasmid library of S. hyodysenteriae isolate B204 genomic DNA in Escherichia coli by colony immunoblot with the mouse monoclonal antibody 10G6/G10, which was produced against cell-free supernatant antigens from the same isolate. Southern blot analysis of HindIII-digested genomic DNA of S. hyodysenteriae serotypes 1 through 7 and of four weakly beta-hemolytic intestinal spirochetes, including Serpulina innocens, with the 2.3-kb DNA fragment of pRED3C6 indicated that the cloned sequence was present exclusively in the seven serotypes of S. hyodysenteriae. An oligonucleotide primer pair for PCR amplification of a 1.55-kb fragment and an internal oligonucleotide probe were designed and synthesized on the basis of sequence analysis of the 2.3-kb DNA fragment of pRED3C6. Purified genomic DNAs from reference isolates of S. hyodysenteriae serotypes 1 through 9, S. innocens, weakly beta-hemolytic intestinal spirochetes belonging to genotypic groups distinct from those of reference Serpulina spp., other cultivable reference isolates of the order Spirochaetales, and enteric bacteria including Escherichia coli, Salmonella spp., Campylobacter spp., and Bacteroides vulgatus were amplified with the oligonucleotide primer pair in a hot-start PCR. The 1.55-kb products were obtained only in the presence of genomic DNA from each of the nine serotypes of S. hyodysenteriae. The specificity of the 1.55-kb products for S. hyodysenteriae was confirmed on the basis of production of a restriction endonuclease pattern of the PCR products identical to the predicted restriction map analysis of pRED3C6 and positive hybridization signal with the S. hyodysenteriae-specific internal oligonucleotide probe. By using total DNA obtained from normal swine feces inoculated with decreasing concentrations of S. hyodysenteriae cells, the sensitivity of the PCR assay was calculated to be between 1 and 10 organisms per 0.1 g of feces. The PCR assay was 1,000 times more sensitive than conventional culture of dysenteric feces on selective medium. There was complete agreement between the results of PCR assays and anaerobic culture on selective agar medium with diagnostic specimen (n = 9) obtained from six farms on which there were cases with clinical signs suggestive of swine dysentery. Detection of S. hyodysenteriae by PCR amplification of DNA has great potential for rapid identification of S. hyodysenteriae in diagnostic specimens.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1195397, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-120726, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1441137, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1515490, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1518039, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1704013, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1746104, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1757550, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1761697, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1882134, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-1911990, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-2219663, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-2336787, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-2644292, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-2745690, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-3218078, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-3235663, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-3539994, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-3771757, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-3782455, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-473537, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-505906, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-6219389, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-7193071, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-7376384, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-8296452, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-8349750, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-8383375, http://linkedlifedata.com/resource/pubmed/commentcorrection/8077394-844908
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0095-1137
pubmed:author
pubmed:issnType
Print
pubmed:volume
32
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1497-502
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
1994
pubmed:articleTitle
Rapid detection of Serpulina hyodysenteriae in diagnostic specimens by PCR.
pubmed:affiliation
Department of Veterinary and Biomedical Sciences, University of Nebraska, Lincoln 68583-0905.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S.