File Download
  Links for fulltext
     (May Require Subscription)
Supplementary

Article: An enzyme-linked immunosorbent assay to detect PCR products of the rfbS gene from serogroup D salmonellae: A rapid screening prototype

TitleAn enzyme-linked immunosorbent assay to detect PCR products of the rfbS gene from serogroup D salmonellae: A rapid screening prototype
Authors
Issue Date1997
PublisherAmerican Society for Microbiology.
Citation
Journal Of Clinical Microbiology, 1997, v. 35 n. 3, p. 714-718 How to Cite?
AbstractWe describe a digoxigenin-based enzyme-linked immunosorbent assay (DIG- ELISA) following a PCR to detect the amplified lipopolysaccharide rfbS gene as a means for rapid screening of serogroup D salmonellae in stool specimens. For pure bacterial cultures, the sensitivity of the PCR DIG-ELISA was approximately 10 bacteria. In the presence of stool materials, the salmonellae were first isolated by an immunomagnetic separation technique with an O9-specific monoclonal antibody, MATy-O9, followed by PCR and DIG- ELISA. The corresponding sensitivity was about 10 to 100 bacteria. To evaluate the assay performance clinically, 203 stool samples from patients with diarrhea were subjected to the routine culture techniques and the PCR ELISA method with overnight enrichment. The conventional culture method identified 145 salmonellae (31 serogroup B, 27 serogroup C, 82 serogroup D, and 5 serogroup E isolates) and 58 non-salmonella bacteria. The PCR ELISA method correctly identified all 82 serogroup D salmonellae (A405 by ELISA, 2.54 ± 0.74) but was negative for the other Salmonella serogroups (A405, 0.26 ± 0.08; n = 63) and non-Salmonella isolates (A405, 0.16 ± 0.04; n = 58). In order to obtain a visible result, the assay takes approximately 6 h (PCR, 4 h; ELISA, 2 h), along with brief enrichment cultivation of the samples (from 4 to 16 h). Thus, the PCR DIG-ELISA offers a fast, accurate, semiquantitative means of detecting infectious agents such as salmonellae, and future robotic automation is possible.
Persistent Identifierhttp://hdl.handle.net/10722/49318
ISSN
2015 Impact Factor: 3.631
2015 SCImago Journal Rankings: 2.151
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLuk, JMen_HK
dc.contributor.authorKongmuang, Uen_HK
dc.contributor.authorTsang, RSWen_HK
dc.contributor.authorLindberg, AAen_HK
dc.date.accessioned2008-06-12T06:39:21Z-
dc.date.available2008-06-12T06:39:21Z-
dc.date.issued1997en_HK
dc.identifier.citationJournal Of Clinical Microbiology, 1997, v. 35 n. 3, p. 714-718en_HK
dc.identifier.issn0095-1137en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49318-
dc.description.abstractWe describe a digoxigenin-based enzyme-linked immunosorbent assay (DIG- ELISA) following a PCR to detect the amplified lipopolysaccharide rfbS gene as a means for rapid screening of serogroup D salmonellae in stool specimens. For pure bacterial cultures, the sensitivity of the PCR DIG-ELISA was approximately 10 bacteria. In the presence of stool materials, the salmonellae were first isolated by an immunomagnetic separation technique with an O9-specific monoclonal antibody, MATy-O9, followed by PCR and DIG- ELISA. The corresponding sensitivity was about 10 to 100 bacteria. To evaluate the assay performance clinically, 203 stool samples from patients with diarrhea were subjected to the routine culture techniques and the PCR ELISA method with overnight enrichment. The conventional culture method identified 145 salmonellae (31 serogroup B, 27 serogroup C, 82 serogroup D, and 5 serogroup E isolates) and 58 non-salmonella bacteria. The PCR ELISA method correctly identified all 82 serogroup D salmonellae (A405 by ELISA, 2.54 ± 0.74) but was negative for the other Salmonella serogroups (A405, 0.26 ± 0.08; n = 63) and non-Salmonella isolates (A405, 0.16 ± 0.04; n = 58). In order to obtain a visible result, the assay takes approximately 6 h (PCR, 4 h; ELISA, 2 h), along with brief enrichment cultivation of the samples (from 4 to 16 h). Thus, the PCR DIG-ELISA offers a fast, accurate, semiquantitative means of detecting infectious agents such as salmonellae, and future robotic automation is possible.en_HK
dc.format.extent418 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherAmerican Society for Microbiology.en_HK
dc.relation.ispartofJournal of Clinical Microbiologyen_HK
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.en_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.rightsCopyright © American Society for Microbiology, Journal of Clinical Microbiology, 1997, v. 35 n. 3, p. 714-718en_HK
dc.subject.meshEnzyme-Linked Immunosorbent Assay - methods - statistics & numerical dataen_HK
dc.subject.meshGenes, Bacterialen_HK
dc.subject.meshPolymerase Chain Reaction - methods - statistics & numerical dataen_HK
dc.subject.meshSalmonella - classification - genetics - isolation & purificationen_HK
dc.subject.meshSalmonella Food Poisoning - diagnosis - microbiologyen_HK
dc.titleAn enzyme-linked immunosorbent assay to detect PCR products of the rfbS gene from serogroup D salmonellae: A rapid screening prototypeen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0095-1137&volume=35&issue=3&spage=714&epage=718&date=1997&atitle=An+enzyme-linked+immunosorbent+assay+to+detect+PCR+products+of+the+rfbS+gene+from+serogroup+D+salmonellae:+a+rapid+screening+prototypeen_HK
dc.identifier.emailLuk, JM: jmluk@hkucc.hku.hken_HK
dc.identifier.authorityLuk, JM=rp00349en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.pmid9041418-
dc.identifier.pmcidPMC229656-
dc.identifier.scopuseid_2-s2.0-0031044821en_HK
dc.identifier.hkuros28334-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0031044821&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume35en_HK
dc.identifier.issue3en_HK
dc.identifier.spage714en_HK
dc.identifier.epage718en_HK
dc.identifier.isiWOS:A1997WJ84800035-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridLuk, JM=7006777791en_HK
dc.identifier.scopusauthoridKongmuang, U=6602328523en_HK
dc.identifier.scopusauthoridTsang, RSW=7102940066en_HK
dc.identifier.scopusauthoridLindberg, AA=7101776377en_HK

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats