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Article: Detection of human papillomavirus in cervical carcinoma: Comparison of peroxidase, Nanogold, and catalyzed reporter deposition (CARD)-Nanogold in situ hybridization

TitleDetection of human papillomavirus in cervical carcinoma: Comparison of peroxidase, Nanogold, and catalyzed reporter deposition (CARD)-Nanogold in situ hybridization
Authors
Issue Date1999
PublisherNature Publishing Group. The Journal's web site is located at http://www.nature.com/modpathol/
Citation
Modern Pathology, 1999, v. 12 n. 7, p. 689-696 How to Cite?
AbstractWe compared three in situ hybridization (ISH) methods for their applicability and sensitivity in detecting human papillomavirus (HPV) in 61 cases (1 Grade 1, 18 Grade 2, 42 Grade 3) of routinely processed squamous cell cervical carcinoma. A commercially available biotinylated probe for HPV- 16/18 was applied to serial sections and detected by conventional streptavidin-biotin-peroxidase ISH, streptavidin-Nanogold-silver ISH, and catalyzed reporter deposition (CARD)-Nanogold-gold ISH. The latter method involved signal amplification by peroxidase-catalyzed deposition of biotinylated tyramides at the hybridization sites, followed by detection of accumulated biotin by streptavidin-Nanogold made visible by autometallography. The HPV-16/18 detection rates for the three methods were 39.3, 44.3, and 65.6%, respectively. In all of the three ISH methods, a punctate staining pattern (single or multiple intranuclear spots of variable size), presumably indicating viral integration, was highly predominant among the positive cases. Two of the cases identified as positive by streptavidin- biotin-peroxidase ISH were rated negative with streptavidin-Nanogold-silver ISH, whereas six cases that were clearly negative with streptavidin-biotin- peroxidase ISH became positively stained with streptavidin-Nanogold ISH. All of these discordant cases were positive by the highly sensitive CARD Nanogold-gold ISH. In addition, the high detection sensitivity of CARD- Nanogold-gold ISH was confirmed by its ability to detect single copies of HPV-16 in SiHa cells. In general, we found that the intense black reaction product from Nanogold autometallography gave superior contrast to that obtained with the peroxidase system. After tyramide signal amplification, the staining was so clearly visible that preparations could be readily screened under low magnification. Our findings precisely demonstrated the need for improved sensitivity in the in situ detection of HPV. The CARD-Nanogold-gold technology looks promising as a highly sensitive method for routine ISH in molecular pathology.
Persistent Identifierhttp://hdl.handle.net/10722/67865
ISSN
2015 Impact Factor: 5.485
2015 SCImago Journal Rankings: 2.803
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorCheung, ALMen_HK
dc.contributor.authorGraf, AHen_HK
dc.contributor.authorHauserKronberger, Cen_HK
dc.contributor.authorDietze, Oen_HK
dc.contributor.authorTubbs, RRen_HK
dc.contributor.authorHacker, GWen_HK
dc.date.accessioned2010-09-06T05:58:59Z-
dc.date.available2010-09-06T05:58:59Z-
dc.date.issued1999en_HK
dc.identifier.citationModern Pathology, 1999, v. 12 n. 7, p. 689-696en_HK
dc.identifier.issn0893-3952en_HK
dc.identifier.urihttp://hdl.handle.net/10722/67865-
dc.description.abstractWe compared three in situ hybridization (ISH) methods for their applicability and sensitivity in detecting human papillomavirus (HPV) in 61 cases (1 Grade 1, 18 Grade 2, 42 Grade 3) of routinely processed squamous cell cervical carcinoma. A commercially available biotinylated probe for HPV- 16/18 was applied to serial sections and detected by conventional streptavidin-biotin-peroxidase ISH, streptavidin-Nanogold-silver ISH, and catalyzed reporter deposition (CARD)-Nanogold-gold ISH. The latter method involved signal amplification by peroxidase-catalyzed deposition of biotinylated tyramides at the hybridization sites, followed by detection of accumulated biotin by streptavidin-Nanogold made visible by autometallography. The HPV-16/18 detection rates for the three methods were 39.3, 44.3, and 65.6%, respectively. In all of the three ISH methods, a punctate staining pattern (single or multiple intranuclear spots of variable size), presumably indicating viral integration, was highly predominant among the positive cases. Two of the cases identified as positive by streptavidin- biotin-peroxidase ISH were rated negative with streptavidin-Nanogold-silver ISH, whereas six cases that were clearly negative with streptavidin-biotin- peroxidase ISH became positively stained with streptavidin-Nanogold ISH. All of these discordant cases were positive by the highly sensitive CARD Nanogold-gold ISH. In addition, the high detection sensitivity of CARD- Nanogold-gold ISH was confirmed by its ability to detect single copies of HPV-16 in SiHa cells. In general, we found that the intense black reaction product from Nanogold autometallography gave superior contrast to that obtained with the peroxidase system. After tyramide signal amplification, the staining was so clearly visible that preparations could be readily screened under low magnification. Our findings precisely demonstrated the need for improved sensitivity in the in situ detection of HPV. The CARD-Nanogold-gold technology looks promising as a highly sensitive method for routine ISH in molecular pathology.en_HK
dc.languageengen_HK
dc.publisherNature Publishing Group. The Journal's web site is located at http://www.nature.com/modpathol/en_HK
dc.relation.ispartofModern Pathologyen_HK
dc.subject.meshCarcinoma, Squamous Cell - genetics - pathology - virologyen_HK
dc.subject.meshDNA, Viral - geneticsen_HK
dc.subject.meshFemaleen_HK
dc.subject.meshGold Compoundsen_HK
dc.subject.meshHumansen_HK
dc.subject.meshIn Situ Hybridization - methodsen_HK
dc.subject.meshPapillomaviridae - geneticsen_HK
dc.subject.meshPapillomavirus Infections - virologyen_HK
dc.subject.meshPeroxidaseen_HK
dc.subject.meshSensitivity and Specificityen_HK
dc.subject.meshTumor Cells, Cultureden_HK
dc.subject.meshTumor Virus Infections - virologyen_HK
dc.subject.meshUterine Cervical Neoplasms - genetics - pathology - virologyen_HK
dc.titleDetection of human papillomavirus in cervical carcinoma: Comparison of peroxidase, Nanogold, and catalyzed reporter deposition (CARD)-Nanogold in situ hybridizationen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0893-3952&volume=12&issue=7&spage=689&epage=696&date=1999&atitle=Detection+of+human+papillomavirus+in+cervical+carcinoma:+comparison+of+peroxidase,+nanogold,+and+catalyzed+reporter+deposition+(CARD)-Nanogold+in+situ+hybridizationen_HK
dc.identifier.emailCheung, ALM:lmcheung@hkucc.hku.hken_HK
dc.identifier.authorityCheung, ALM=rp00332en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.pmid10430273-
dc.identifier.scopuseid_2-s2.0-0032868589en_HK
dc.identifier.hkuros46344en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0032868589&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume12en_HK
dc.identifier.issue7en_HK
dc.identifier.spage689en_HK
dc.identifier.epage696en_HK
dc.identifier.isiWOS:000081559700005-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridCheung, ALM=7401806497en_HK
dc.identifier.scopusauthoridGraf, AH=7006625267en_HK
dc.identifier.scopusauthoridHauserKronberger, C=7003360268en_HK
dc.identifier.scopusauthoridDietze, O=24798646300en_HK
dc.identifier.scopusauthoridTubbs, RR=35400790200en_HK
dc.identifier.scopusauthoridHacker, GW=7006705677en_HK

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