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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Veröffentlicht in:Research in microbiology 2011-06, Vol.162 (5), p.499-505
Hauptverfasser: 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
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container_end_page 505
container_issue 5
container_start_page 499
container_title Research in microbiology
container_volume 162
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
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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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