Detection of hydrogen peroxide-producing Lactobacillus species in the vagina

a comparison of culture and quantitative PCR among HIV-1 seropositive women

Jennifer E. Balkus, Caroline Mitchell, Kathy Agnew, Congzhou Liu, Tina Fiedler, Susan E. Cohn, Amneris Luque, Robert Coombs, David N. Fredricks, Jane Hitti

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Background: The presence of hydrogen peroxide (H2O2) producing Lactobacillus in the vagina may play a role in controlling genital HIV-1 shedding. Sensitive molecular methods improve our ability to characterize the vaginal microbiota; however, they cannot characterize phenotype. We assessed the concordance of H2O2-producing Lactobacillus detected by culture with quantitative PCR (qPCR) detection of Lactobacillus species commonly assumed to be H2O2-producers.Methods: Samples were collected as part of a prospective cohort study of HIV-1 seropositive US women. Cervicovaginal lavage specimens were tested for L. crispatus and L. jensenii using 16S rRNA gene qPCR assays. Vaginal swabs were cultured for Lactobacillus and tested for H2O2-production. We calculated a kappa statistic to assess concordance between culture and qPCR.Results: Culture and qPCR results were available for 376 visits from 57 women. Lactobacilli were detected by culture at 308 (82%) visits, of which 233 of 308 (76%) produced H2O2. L. crispatus and/or L. jensenii were detected at 215 (57%) visits. Concordance between detection of L. crispatus and/or L. jensenii by qPCR and H2O2-producing Lactobacillus by culture was 75% (kappa = 0.45).Conclusions: Among HIV-1 seropositive women, there was a moderate level of concordance between H2O2-producing Lactobacillus detected by culture and the presence of L. crispatus and/or L. jensenii by qPCR. However, one-quarter of samples with growth of H2O2-producing lactobacilli did not have L. crispatus or L. jensenii detected by qPCR. This discordance may be due to the presence of other H2O2-producing Lactobacillus species.

Original languageEnglish (US)
Article number188
JournalBMC Infectious Diseases
Volume12
DOIs
StatePublished - Aug 13 2012

Fingerprint

Lactobacillus
Vagina
Hydrogen Peroxide
HIV-1
Polymerase Chain Reaction
Therapeutic Irrigation
Microbiota
rRNA Genes
Cohort Studies
Prospective Studies
Phenotype
Growth

Keywords

  • 16S rRNA gene
  • Bacterial vaginosis
  • Culture
  • Hydrogen peroxide
  • Lactobacillus
  • Lactobacillus crispatus
  • Lactobacillus jensenii

ASJC Scopus subject areas

  • Infectious Diseases

Cite this

Detection of hydrogen peroxide-producing Lactobacillus species in the vagina : a comparison of culture and quantitative PCR among HIV-1 seropositive women. / Balkus, Jennifer E.; Mitchell, Caroline; Agnew, Kathy; Liu, Congzhou; Fiedler, Tina; Cohn, Susan E.; Luque, Amneris; Coombs, Robert; Fredricks, David N.; Hitti, Jane.

In: BMC Infectious Diseases, Vol. 12, 188, 13.08.2012.

Research output: Contribution to journalArticle

Balkus, Jennifer E. ; Mitchell, Caroline ; Agnew, Kathy ; Liu, Congzhou ; Fiedler, Tina ; Cohn, Susan E. ; Luque, Amneris ; Coombs, Robert ; Fredricks, David N. ; Hitti, Jane. / Detection of hydrogen peroxide-producing Lactobacillus species in the vagina : a comparison of culture and quantitative PCR among HIV-1 seropositive women. In: BMC Infectious Diseases. 2012 ; Vol. 12.
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abstract = "Background: The presence of hydrogen peroxide (H2O2) producing Lactobacillus in the vagina may play a role in controlling genital HIV-1 shedding. Sensitive molecular methods improve our ability to characterize the vaginal microbiota; however, they cannot characterize phenotype. We assessed the concordance of H2O2-producing Lactobacillus detected by culture with quantitative PCR (qPCR) detection of Lactobacillus species commonly assumed to be H2O2-producers.Methods: Samples were collected as part of a prospective cohort study of HIV-1 seropositive US women. Cervicovaginal lavage specimens were tested for L. crispatus and L. jensenii using 16S rRNA gene qPCR assays. Vaginal swabs were cultured for Lactobacillus and tested for H2O2-production. We calculated a kappa statistic to assess concordance between culture and qPCR.Results: Culture and qPCR results were available for 376 visits from 57 women. Lactobacilli were detected by culture at 308 (82{\%}) visits, of which 233 of 308 (76{\%}) produced H2O2. L. crispatus and/or L. jensenii were detected at 215 (57{\%}) visits. Concordance between detection of L. crispatus and/or L. jensenii by qPCR and H2O2-producing Lactobacillus by culture was 75{\%} (kappa = 0.45).Conclusions: Among HIV-1 seropositive women, there was a moderate level of concordance between H2O2-producing Lactobacillus detected by culture and the presence of L. crispatus and/or L. jensenii by qPCR. However, one-quarter of samples with growth of H2O2-producing lactobacilli did not have L. crispatus or L. jensenii detected by qPCR. This discordance may be due to the presence of other H2O2-producing Lactobacillus species.",
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AU - Mitchell, Caroline

AU - Agnew, Kathy

AU - Liu, Congzhou

AU - Fiedler, Tina

AU - Cohn, Susan E.

AU - Luque, Amneris

AU - Coombs, Robert

AU - Fredricks, David N.

AU - Hitti, Jane

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N2 - Background: The presence of hydrogen peroxide (H2O2) producing Lactobacillus in the vagina may play a role in controlling genital HIV-1 shedding. Sensitive molecular methods improve our ability to characterize the vaginal microbiota; however, they cannot characterize phenotype. We assessed the concordance of H2O2-producing Lactobacillus detected by culture with quantitative PCR (qPCR) detection of Lactobacillus species commonly assumed to be H2O2-producers.Methods: Samples were collected as part of a prospective cohort study of HIV-1 seropositive US women. Cervicovaginal lavage specimens were tested for L. crispatus and L. jensenii using 16S rRNA gene qPCR assays. Vaginal swabs were cultured for Lactobacillus and tested for H2O2-production. We calculated a kappa statistic to assess concordance between culture and qPCR.Results: Culture and qPCR results were available for 376 visits from 57 women. Lactobacilli were detected by culture at 308 (82%) visits, of which 233 of 308 (76%) produced H2O2. L. crispatus and/or L. jensenii were detected at 215 (57%) visits. Concordance between detection of L. crispatus and/or L. jensenii by qPCR and H2O2-producing Lactobacillus by culture was 75% (kappa = 0.45).Conclusions: Among HIV-1 seropositive women, there was a moderate level of concordance between H2O2-producing Lactobacillus detected by culture and the presence of L. crispatus and/or L. jensenii by qPCR. However, one-quarter of samples with growth of H2O2-producing lactobacilli did not have L. crispatus or L. jensenii detected by qPCR. This discordance may be due to the presence of other H2O2-producing Lactobacillus species.

AB - Background: The presence of hydrogen peroxide (H2O2) producing Lactobacillus in the vagina may play a role in controlling genital HIV-1 shedding. Sensitive molecular methods improve our ability to characterize the vaginal microbiota; however, they cannot characterize phenotype. We assessed the concordance of H2O2-producing Lactobacillus detected by culture with quantitative PCR (qPCR) detection of Lactobacillus species commonly assumed to be H2O2-producers.Methods: Samples were collected as part of a prospective cohort study of HIV-1 seropositive US women. Cervicovaginal lavage specimens were tested for L. crispatus and L. jensenii using 16S rRNA gene qPCR assays. Vaginal swabs were cultured for Lactobacillus and tested for H2O2-production. We calculated a kappa statistic to assess concordance between culture and qPCR.Results: Culture and qPCR results were available for 376 visits from 57 women. Lactobacilli were detected by culture at 308 (82%) visits, of which 233 of 308 (76%) produced H2O2. L. crispatus and/or L. jensenii were detected at 215 (57%) visits. Concordance between detection of L. crispatus and/or L. jensenii by qPCR and H2O2-producing Lactobacillus by culture was 75% (kappa = 0.45).Conclusions: Among HIV-1 seropositive women, there was a moderate level of concordance between H2O2-producing Lactobacillus detected by culture and the presence of L. crispatus and/or L. jensenii by qPCR. However, one-quarter of samples with growth of H2O2-producing lactobacilli did not have L. crispatus or L. jensenii detected by qPCR. This discordance may be due to the presence of other H2O2-producing Lactobacillus species.

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KW - Bacterial vaginosis

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KW - Hydrogen peroxide

KW - Lactobacillus

KW - Lactobacillus crispatus

KW - Lactobacillus jensenii

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