Novel Short-Fragment PCR Assay for Highly Sensitive Broad-Spectrum Detection of Anogenital Human Papillomaviruses
A novel set of polymerase chain reaction (PCR) primers, designated SPF1 and SPF2 and located in the L1 region, was developed for universal detection of human papillomavirus (HPV). A short PCR fragment (SPF) of only 65 pb was synthesized. SPF amplimers were detected in a microtiter-based hybridizatio...
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| Veröffentlicht in: | The American journal of pathology 1998-12, Vol.153 (6), p.1731-1739 |
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| container_title | The American journal of pathology |
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| creator | Kleter, Bernhard van Doorn, Leen-Jan ter Schegget, Jan Schrauwen, Lianne van Krimpen, Kees Burger, Matthé ter Harmsel, Bram Quint, Wim |
| description | A novel set of polymerase chain reaction (PCR) primers, designated SPF1 and SPF2 and located in the L1 region, was developed for universal detection of human papillomavirus (HPV). A short PCR fragment (SPF) of only 65 pb was synthesized. SPF amplimers were detected in a microtiter-based hybridization system, using a mixture of oligonucleotide probes. The SPF system allowed detection of at least 43 different HPV genotypes. The clinical performance of the novel SPF system was assessed in three different patient groups. 1) Analysis of 534 cervical scrapes, obtained from treated patients, showed that the detection rate in 447 (83.7%) scrapes with normal cytology was significantly higher using the SPF system as compared with the universal primer set GP5+/6+ (
P < 0.001). 2) The SPF assay detected HPV DNA in 299 (98.4%) of 304 scrapes with cytological dyskaryosis. 3) The SPF system detected HPV DNA in 100% of 184 formalin-fixed, paraffin-embedded cervical carcinoma specimens. In conclusion, the novel SPF system permitted universal and highly sensitive detection of HPV DNA in diverse clinical materials and may improve the molecular diagnosis and epidemiology of this important virus. |
| doi_str_mv | 10.1016/S0002-9440(10)65688-X |
| format | Article |
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P < 0.001). 2) The SPF assay detected HPV DNA in 299 (98.4%) of 304 scrapes with cytological dyskaryosis. 3) The SPF system detected HPV DNA in 100% of 184 formalin-fixed, paraffin-embedded cervical carcinoma specimens. In conclusion, the novel SPF system permitted universal and highly sensitive detection of HPV DNA in diverse clinical materials and may improve the molecular diagnosis and epidemiology of this important virus.</description><identifier>ISSN: 0002-9440</identifier><identifier>EISSN: 1525-2191</identifier><identifier>DOI: 10.1016/S0002-9440(10)65688-X</identifier><identifier>PMID: 9846964</identifier><identifier>CODEN: AJPAA4</identifier><language>eng</language><publisher>Bethesda, MD: Elsevier Inc</publisher><subject>Biological and medical sciences ; Blotting, Southern ; Cervix Uteri - virology ; Female ; Humans ; Investigative techniques, diagnostic techniques (general aspects) ; Medical sciences ; Miscellaneous. Technology ; Oligonucleotide Probes ; Papillomaviridae - isolation & purification ; Papillomavirus Infections - diagnosis ; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques ; Polymerase Chain Reaction - methods ; Precancerous Conditions - virology ; Sensitivity and Specificity ; Technical Advance ; Tumor Virus Infections - diagnosis ; Uterine Cervical Neoplasms - virology ; Vaginal Smears</subject><ispartof>The American journal of pathology, 1998-12, Vol.153 (6), p.1731-1739</ispartof><rights>1998 American Society for Investigative Pathology</rights><rights>1999 INIST-CNRS</rights><rights>Copyright American Society for Investigative Pathology Dec 1998</rights><rights>Copyright © 1998, American Society for Investigative Pathology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c602t-3d4f7e5d9e93d95f84cc079a8a726451132a966cf61c319fa33ccba357011ea23</citedby><cites>FETCH-LOGICAL-c602t-3d4f7e5d9e93d95f84cc079a8a726451132a966cf61c319fa33ccba357011ea23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1866345/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S000294401065688X$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,3537,27901,27902,53766,53768,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1607467$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9846964$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kleter, Bernhard</creatorcontrib><creatorcontrib>van Doorn, Leen-Jan</creatorcontrib><creatorcontrib>ter Schegget, Jan</creatorcontrib><creatorcontrib>Schrauwen, Lianne</creatorcontrib><creatorcontrib>van Krimpen, Kees</creatorcontrib><creatorcontrib>Burger, Matthé</creatorcontrib><creatorcontrib>ter Harmsel, Bram</creatorcontrib><creatorcontrib>Quint, Wim</creatorcontrib><title>Novel Short-Fragment PCR Assay for Highly Sensitive Broad-Spectrum Detection of Anogenital Human Papillomaviruses</title><title>The American journal of pathology</title><addtitle>Am J Pathol</addtitle><description>A novel set of polymerase chain reaction (PCR) primers, designated SPF1 and SPF2 and located in the L1 region, was developed for universal detection of human papillomavirus (HPV). A short PCR fragment (SPF) of only 65 pb was synthesized. SPF amplimers were detected in a microtiter-based hybridization system, using a mixture of oligonucleotide probes. The SPF system allowed detection of at least 43 different HPV genotypes. The clinical performance of the novel SPF system was assessed in three different patient groups. 1) Analysis of 534 cervical scrapes, obtained from treated patients, showed that the detection rate in 447 (83.7%) scrapes with normal cytology was significantly higher using the SPF system as compared with the universal primer set GP5+/6+ (
P < 0.001). 2) The SPF assay detected HPV DNA in 299 (98.4%) of 304 scrapes with cytological dyskaryosis. 3) The SPF system detected HPV DNA in 100% of 184 formalin-fixed, paraffin-embedded cervical carcinoma specimens. In conclusion, the novel SPF system permitted universal and highly sensitive detection of HPV DNA in diverse clinical materials and may improve the molecular diagnosis and epidemiology of this important virus.</description><subject>Biological and medical sciences</subject><subject>Blotting, Southern</subject><subject>Cervix Uteri - virology</subject><subject>Female</subject><subject>Humans</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Medical sciences</subject><subject>Miscellaneous. Technology</subject><subject>Oligonucleotide Probes</subject><subject>Papillomaviridae - isolation & purification</subject><subject>Papillomavirus Infections - diagnosis</subject><subject>Pathology. Cytology. Biochemistry. 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A short PCR fragment (SPF) of only 65 pb was synthesized. SPF amplimers were detected in a microtiter-based hybridization system, using a mixture of oligonucleotide probes. The SPF system allowed detection of at least 43 different HPV genotypes. The clinical performance of the novel SPF system was assessed in three different patient groups. 1) Analysis of 534 cervical scrapes, obtained from treated patients, showed that the detection rate in 447 (83.7%) scrapes with normal cytology was significantly higher using the SPF system as compared with the universal primer set GP5+/6+ (
P < 0.001). 2) The SPF assay detected HPV DNA in 299 (98.4%) of 304 scrapes with cytological dyskaryosis. 3) The SPF system detected HPV DNA in 100% of 184 formalin-fixed, paraffin-embedded cervical carcinoma specimens. In conclusion, the novel SPF system permitted universal and highly sensitive detection of HPV DNA in diverse clinical materials and may improve the molecular diagnosis and epidemiology of this important virus.</abstract><cop>Bethesda, MD</cop><pub>Elsevier Inc</pub><pmid>9846964</pmid><doi>10.1016/S0002-9440(10)65688-X</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Biological and medical sciences Blotting, Southern Cervix Uteri - virology Female Humans Investigative techniques, diagnostic techniques (general aspects) Medical sciences Miscellaneous. Technology Oligonucleotide Probes Papillomaviridae - isolation & purification Papillomavirus Infections - diagnosis Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Polymerase Chain Reaction - methods Precancerous Conditions - virology Sensitivity and Specificity Technical Advance Tumor Virus Infections - diagnosis Uterine Cervical Neoplasms - virology Vaginal Smears |
| title | Novel Short-Fragment PCR Assay for Highly Sensitive Broad-Spectrum Detection of Anogenital Human Papillomaviruses |
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