Comparison of culture with two different qPCR assays for detection of rectovaginal carriage of Streptococcus agalactiae (group B streptococci) in pregnant women
Development of rapid and sensitive detection methods for group B streptococci (GBS) in pregnant women remains useful in order to adequately identify pregnant women at risk of transferring GBS to their neonate. This study compared the CDC recommended sampling and culture method with two qPCR methods...
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creator | El Aila, Nabil Abdullah Tency, Inge Claeys, Geert Verstraelen, Hans Deschaght, Pieter Decat, Ellen Lopes dos Santos Santiago, Guido Cools, Piet Temmerman, Marleen Vaneechoutte, Mario |
description | Development of rapid and sensitive detection methods for group B streptococci (GBS) in pregnant women remains useful in order to adequately identify pregnant women at risk of transferring GBS to their neonate. This study compared the CDC recommended sampling and culture method with two qPCR methods for detecting GBS colonization.
For a total of 100 pregnant women at 35–37 weeks of gestation, one rectovaginal ESwab each was collected. Eswab medium was inoculated into Lim broth, incubated for 24 h and plated onto chromID™ Strepto B agar (ChromAgar). DNA was extracted with the bioMérieux easyMAG platform, either directly from the rectovaginal ESwab or from Lim broth enrichment culture. Two different qPCR formats were compared, i.e. the hydrolysis probe format (Taqman, Roche) targeting the
sip gene and the hybridization probe format (Hybprobe, Roche) targeting the
cfb gene.
Both qPCR techniques identified 33% of the women as GBS-positive. Only one culture-positive sample was qPCR-negative. QPCR directly on the sample significantly increased the number of women found to be GBS-positive (27%) compared to culture (22%). Moreover, the sensitivity of qPCR after Lim broth enrichment (33%) was again significantly higher than qPCR after DNA extraction directly from the rectovaginal swabs (27%).
In conclusion, for prenatal screening of GBS from rectovaginal samples of pregnant women, our results are in accordance with CDC guidelines, which suggest using qPCR after Lim broth enrichment in addition to conventional (culture-based) detection. qPCR after Lim broth enrichment further increased the percentage of GBS-positive women, as detected by direct qPCR, from 27 to 33%, although the bacterial inoculum was low for these subjects. |
doi_str_mv | 10.1016/j.resmic.2011.04.001 |
format | Article |
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For a total of 100 pregnant women at 35–37 weeks of gestation, one rectovaginal ESwab each was collected. Eswab medium was inoculated into Lim broth, incubated for 24 h and plated onto chromID™ Strepto B agar (ChromAgar). DNA was extracted with the bioMérieux easyMAG platform, either directly from the rectovaginal ESwab or from Lim broth enrichment culture. Two different qPCR formats were compared, i.e. the hydrolysis probe format (Taqman, Roche) targeting the
sip gene and the hybridization probe format (Hybprobe, Roche) targeting the
cfb gene.
Both qPCR techniques identified 33% of the women as GBS-positive. Only one culture-positive sample was qPCR-negative. QPCR directly on the sample significantly increased the number of women found to be GBS-positive (27%) compared to culture (22%). Moreover, the sensitivity of qPCR after Lim broth enrichment (33%) was again significantly higher than qPCR after DNA extraction directly from the rectovaginal swabs (27%).
In conclusion, for prenatal screening of GBS from rectovaginal samples of pregnant women, our results are in accordance with CDC guidelines, which suggest using qPCR after Lim broth enrichment in addition to conventional (culture-based) detection. qPCR after Lim broth enrichment further increased the percentage of GBS-positive women, as detected by direct qPCR, from 27 to 33%, although the bacterial inoculum was low for these subjects.</description><identifier>ISSN: 0923-2508</identifier><identifier>EISSN: 1769-7123</identifier><identifier>DOI: 10.1016/j.resmic.2011.04.001</identifier><identifier>PMID: 21514378</identifier><language>eng</language><publisher>Issy-les-Moulineaux: Elsevier SAS</publisher><subject>Agar ; Bacteriology ; Biological and medical sciences ; Chromagar ; Colonization ; Culture Media ; DNA ; Female ; Fundamental and applied biological sciences. Psychology ; Gestation ; Group B streptococci ; Humans ; Hydrolysis ; Inoculum ; Lim broth ; Microbiology ; Miscellaneous ; Neonates ; Polymerase Chain Reaction - methods ; Pregnancy ; Pregnancy Complications, Infectious - diagnosis ; Pregnancy Complications, Infectious - microbiology ; Pregnant women ; Probes ; qPCR ; Rectum - microbiology ; Sampling ; sip gene ; Streptococcal Infections - diagnosis ; Streptococcal Infections - microbiology ; Streptococcus agalactiae ; Streptococcus agalactiae - genetics ; Streptococcus agalactiae - growth & development ; Streptococcus agalactiae - isolation & purification ; Vagina - microbiology</subject><ispartof>Research in microbiology, 2011-06, Vol.162 (5), p.499-505</ispartof><rights>2011 Institut Pasteur</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2011 Institut Pasteur. Published by Elsevier SAS. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c493t-f3e254c7eeaa810c0707d6db63ffbb923f9ee55347780d0b4cf4bc75c9a4c313</citedby><cites>FETCH-LOGICAL-c493t-f3e254c7eeaa810c0707d6db63ffbb923f9ee55347780d0b4cf4bc75c9a4c313</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.resmic.2011.04.001$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,27923,27924,45994</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24303466$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21514378$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>El Aila, Nabil Abdullah</creatorcontrib><creatorcontrib>Tency, Inge</creatorcontrib><creatorcontrib>Claeys, Geert</creatorcontrib><creatorcontrib>Verstraelen, Hans</creatorcontrib><creatorcontrib>Deschaght, Pieter</creatorcontrib><creatorcontrib>Decat, Ellen</creatorcontrib><creatorcontrib>Lopes dos Santos Santiago, Guido</creatorcontrib><creatorcontrib>Cools, Piet</creatorcontrib><creatorcontrib>Temmerman, Marleen</creatorcontrib><creatorcontrib>Vaneechoutte, Mario</creatorcontrib><title>Comparison of culture with two different qPCR assays for detection of rectovaginal carriage of Streptococcus agalactiae (group B streptococci) in pregnant women</title><title>Research in microbiology</title><addtitle>Res Microbiol</addtitle><description>Development of rapid and sensitive detection methods for group B streptococci (GBS) in pregnant women remains useful in order to adequately identify pregnant women at risk of transferring GBS to their neonate. This study compared the CDC recommended sampling and culture method with two qPCR methods for detecting GBS colonization.
For a total of 100 pregnant women at 35–37 weeks of gestation, one rectovaginal ESwab each was collected. Eswab medium was inoculated into Lim broth, incubated for 24 h and plated onto chromID™ Strepto B agar (ChromAgar). DNA was extracted with the bioMérieux easyMAG platform, either directly from the rectovaginal ESwab or from Lim broth enrichment culture. Two different qPCR formats were compared, i.e. the hydrolysis probe format (Taqman, Roche) targeting the
sip gene and the hybridization probe format (Hybprobe, Roche) targeting the
cfb gene.
Both qPCR techniques identified 33% of the women as GBS-positive. Only one culture-positive sample was qPCR-negative. QPCR directly on the sample significantly increased the number of women found to be GBS-positive (27%) compared to culture (22%). Moreover, the sensitivity of qPCR after Lim broth enrichment (33%) was again significantly higher than qPCR after DNA extraction directly from the rectovaginal swabs (27%).
In conclusion, for prenatal screening of GBS from rectovaginal samples of pregnant women, our results are in accordance with CDC guidelines, which suggest using qPCR after Lim broth enrichment in addition to conventional (culture-based) detection. qPCR after Lim broth enrichment further increased the percentage of GBS-positive women, as detected by direct qPCR, from 27 to 33%, although the bacterial inoculum was low for these subjects.</description><subject>Agar</subject><subject>Bacteriology</subject><subject>Biological and medical sciences</subject><subject>Chromagar</subject><subject>Colonization</subject><subject>Culture Media</subject><subject>DNA</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gestation</subject><subject>Group B streptococci</subject><subject>Humans</subject><subject>Hydrolysis</subject><subject>Inoculum</subject><subject>Lim broth</subject><subject>Microbiology</subject><subject>Miscellaneous</subject><subject>Neonates</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Pregnancy</subject><subject>Pregnancy Complications, Infectious - diagnosis</subject><subject>Pregnancy Complications, Infectious - microbiology</subject><subject>Pregnant women</subject><subject>Probes</subject><subject>qPCR</subject><subject>Rectum - microbiology</subject><subject>Sampling</subject><subject>sip gene</subject><subject>Streptococcal Infections - diagnosis</subject><subject>Streptococcal Infections - microbiology</subject><subject>Streptococcus agalactiae</subject><subject>Streptococcus agalactiae - genetics</subject><subject>Streptococcus agalactiae - growth & development</subject><subject>Streptococcus agalactiae - isolation & purification</subject><subject>Vagina - microbiology</subject><issn>0923-2508</issn><issn>1769-7123</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1u1DAURiMEokPhDRB4g4DFBDt24mSDBCP-pEogWtbWjXMdPEri1HY66tvwqHiUge5gZUs-n7-re7LsKaM5o6x6s889htHqvKCM5VTklLJ72YbJqtlKVvD72YY2Bd8WJa3Pskch7BNQSikeZmcFK5ngst5kv3ZunMHb4CbiDNHLEBeP5GDjTxIPjnTWGPQ4RXL9bfedQAhwG4hxnnQYUUe75ny6uhvo7QQD0eC9hR6PD5fR4xyddlovgUAPA6QQIHnVe7fM5D0Jd4R9TexEZo_9BKnx4EacHmcPDAwBn5zO8-zq44er3eftxddPX3bvLrZaNDxuDceiFFoiAtSMaiqp7KqurbgxbZv2YBrEsuRCypp2tBXaiFbLUjcgNGf8PHu5fjt7d71giGq0QeMwwIRuCapuGiYYq8X_SUlTz0qKldTeheDRqNnbEfytYlQdHaq9Wh2qo0NFhUqKUuzZqWBpR-z-hv5IS8CLEwBBw2A8TNqGO05wykVVJe75yhlwCvokWf24TE1VapFpSpqItyuBabM3Fr0K2uKksbNHo6pz9t-z_gYSvsi4</recordid><startdate>20110601</startdate><enddate>20110601</enddate><creator>El Aila, Nabil Abdullah</creator><creator>Tency, Inge</creator><creator>Claeys, Geert</creator><creator>Verstraelen, Hans</creator><creator>Deschaght, Pieter</creator><creator>Decat, Ellen</creator><creator>Lopes dos Santos Santiago, Guido</creator><creator>Cools, Piet</creator><creator>Temmerman, Marleen</creator><creator>Vaneechoutte, Mario</creator><general>Elsevier SAS</general><general>Elsevier Masson</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7QL</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>P64</scope></search><sort><creationdate>20110601</creationdate><title>Comparison of culture with two different qPCR assays for detection of rectovaginal carriage of Streptococcus agalactiae (group B streptococci) in pregnant women</title><author>El Aila, Nabil Abdullah ; Tency, Inge ; Claeys, Geert ; Verstraelen, Hans ; Deschaght, Pieter ; Decat, Ellen ; Lopes dos Santos Santiago, Guido ; Cools, Piet ; Temmerman, Marleen ; Vaneechoutte, Mario</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c493t-f3e254c7eeaa810c0707d6db63ffbb923f9ee55347780d0b4cf4bc75c9a4c313</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Agar</topic><topic>Bacteriology</topic><topic>Biological and medical sciences</topic><topic>Chromagar</topic><topic>Colonization</topic><topic>Culture Media</topic><topic>DNA</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gestation</topic><topic>Group B streptococci</topic><topic>Humans</topic><topic>Hydrolysis</topic><topic>Inoculum</topic><topic>Lim broth</topic><topic>Microbiology</topic><topic>Miscellaneous</topic><topic>Neonates</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Pregnancy</topic><topic>Pregnancy Complications, Infectious - diagnosis</topic><topic>Pregnancy Complications, Infectious - microbiology</topic><topic>Pregnant women</topic><topic>Probes</topic><topic>qPCR</topic><topic>Rectum - microbiology</topic><topic>Sampling</topic><topic>sip gene</topic><topic>Streptococcal Infections - diagnosis</topic><topic>Streptococcal Infections - microbiology</topic><topic>Streptococcus agalactiae</topic><topic>Streptococcus agalactiae - genetics</topic><topic>Streptococcus agalactiae - growth & development</topic><topic>Streptococcus agalactiae - isolation & purification</topic><topic>Vagina - microbiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>El Aila, Nabil Abdullah</creatorcontrib><creatorcontrib>Tency, Inge</creatorcontrib><creatorcontrib>Claeys, Geert</creatorcontrib><creatorcontrib>Verstraelen, Hans</creatorcontrib><creatorcontrib>Deschaght, Pieter</creatorcontrib><creatorcontrib>Decat, Ellen</creatorcontrib><creatorcontrib>Lopes dos Santos Santiago, Guido</creatorcontrib><creatorcontrib>Cools, Piet</creatorcontrib><creatorcontrib>Temmerman, Marleen</creatorcontrib><creatorcontrib>Vaneechoutte, Mario</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Research in microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>El Aila, Nabil Abdullah</au><au>Tency, Inge</au><au>Claeys, Geert</au><au>Verstraelen, Hans</au><au>Deschaght, Pieter</au><au>Decat, Ellen</au><au>Lopes dos Santos Santiago, Guido</au><au>Cools, Piet</au><au>Temmerman, Marleen</au><au>Vaneechoutte, Mario</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparison of culture with two different qPCR assays for detection of rectovaginal carriage of Streptococcus agalactiae (group B streptococci) in pregnant women</atitle><jtitle>Research in microbiology</jtitle><addtitle>Res Microbiol</addtitle><date>2011-06-01</date><risdate>2011</risdate><volume>162</volume><issue>5</issue><spage>499</spage><epage>505</epage><pages>499-505</pages><issn>0923-2508</issn><eissn>1769-7123</eissn><abstract>Development of rapid and sensitive detection methods for group B streptococci (GBS) in pregnant women remains useful in order to adequately identify pregnant women at risk of transferring GBS to their neonate. This study compared the CDC recommended sampling and culture method with two qPCR methods for detecting GBS colonization.
For a total of 100 pregnant women at 35–37 weeks of gestation, one rectovaginal ESwab each was collected. Eswab medium was inoculated into Lim broth, incubated for 24 h and plated onto chromID™ Strepto B agar (ChromAgar). DNA was extracted with the bioMérieux easyMAG platform, either directly from the rectovaginal ESwab or from Lim broth enrichment culture. Two different qPCR formats were compared, i.e. the hydrolysis probe format (Taqman, Roche) targeting the
sip gene and the hybridization probe format (Hybprobe, Roche) targeting the
cfb gene.
Both qPCR techniques identified 33% of the women as GBS-positive. Only one culture-positive sample was qPCR-negative. QPCR directly on the sample significantly increased the number of women found to be GBS-positive (27%) compared to culture (22%). Moreover, the sensitivity of qPCR after Lim broth enrichment (33%) was again significantly higher than qPCR after DNA extraction directly from the rectovaginal swabs (27%).
In conclusion, for prenatal screening of GBS from rectovaginal samples of pregnant women, our results are in accordance with CDC guidelines, which suggest using qPCR after Lim broth enrichment in addition to conventional (culture-based) detection. qPCR after Lim broth enrichment further increased the percentage of GBS-positive women, as detected by direct qPCR, from 27 to 33%, although the bacterial inoculum was low for these subjects.</abstract><cop>Issy-les-Moulineaux</cop><pub>Elsevier SAS</pub><pmid>21514378</pmid><doi>10.1016/j.resmic.2011.04.001</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Agar Bacteriology Biological and medical sciences Chromagar Colonization Culture Media DNA Female Fundamental and applied biological sciences. Psychology Gestation Group B streptococci Humans Hydrolysis Inoculum Lim broth Microbiology Miscellaneous Neonates Polymerase Chain Reaction - methods Pregnancy Pregnancy Complications, Infectious - diagnosis Pregnancy Complications, Infectious - microbiology Pregnant women Probes qPCR Rectum - microbiology Sampling sip gene Streptococcal Infections - diagnosis Streptococcal Infections - microbiology Streptococcus agalactiae Streptococcus agalactiae - genetics Streptococcus agalactiae - growth & development Streptococcus agalactiae - isolation & purification Vagina - microbiology |
title | Comparison of culture with two different qPCR assays for detection of rectovaginal carriage of Streptococcus agalactiae (group B streptococci) in pregnant women |
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