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

doi:10.1186/1471-2334-12-188

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

^*Corresponding author for this work

Medicine, Infectious Diseases Division

Research output: Contribution to journal › Article › peer-review

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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.

Original language	English (US)
Article number	188
Journal	BMC Infectious Diseases
Volume	12
DOIs	https://doi.org/10.1186/1471-2334-12-188
State	Published - Aug 13 2012

Funding

We gratefully acknowledge support from our clinic staff, laboratory staff and administrators for their dedication and hard work. We sincerely thank the women whose time, effort, and commitment made this research possible. This research was supported by grants from the National Institutes of Health [grant R21 AI080439 and P01 HD040540]. JE Balkus was supported in part by a grant from the University of Washington Center for AIDS and STDs [grant T32 AI007140-32].

Keywords

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

ASJC Scopus subject areas

Infectious Diseases

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1186/1471-2334-12-188

Cite this

Balkus, J. E., Mitchell, C., Agnew, K., Liu, C., Fiedler, T., Cohn, S. E., Luque, A., Coombs, R., Fredricks, D. N., & Hitti, J. (2012). Detection of hydrogen peroxide-producing Lactobacillus species in the vagina: a comparison of culture and quantitative PCR among HIV-1 seropositive women. BMC Infectious Diseases, 12, Article 188. https://doi.org/10.1186/1471-2334-12-188

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title = "Detection of hydrogen peroxide-producing Lactobacillus species in the vagina: a comparison of culture and quantitative PCR among HIV-1 seropositive women",

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.",

keywords = "16S rRNA gene, Bacterial vaginosis, Culture, Hydrogen peroxide, Lactobacillus, Lactobacillus crispatus, Lactobacillus jensenii",

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

note = "Funding Information: We gratefully acknowledge support from our clinic staff, laboratory staff and administrators for their dedication and hard work. We sincerely thank the women whose time, effort, and commitment made this research possible. This research was supported by grants from the National Institutes of Health [grant R21 AI080439 and P01 HD040540]. JE Balkus was supported in part by a grant from the University of Washington Center for AIDS and STDs [grant T32 AI007140-32].",

year = "2012",

month = aug,

day = "13",

doi = "10.1186/1471-2334-12-188",

language = "English (US)",

volume = "12",

journal = "BMC Infectious Diseases",

issn = "1471-2334",

publisher = "BioMed Central Ltd.",

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TY - JOUR

T1 - Detection of hydrogen peroxide-producing Lactobacillus species in the vagina

T2 - a comparison of culture and quantitative PCR among HIV-1 seropositive women

AU - Balkus, Jennifer E.

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

N1 - Funding Information: We gratefully acknowledge support from our clinic staff, laboratory staff and administrators for their dedication and hard work. We sincerely thank the women whose time, effort, and commitment made this research possible. This research was supported by grants from the National Institutes of Health [grant R21 AI080439 and P01 HD040540]. JE Balkus was supported in part by a grant from the University of Washington Center for AIDS and STDs [grant T32 AI007140-32].

PY - 2012/8/13

Y1 - 2012/8/13

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.

KW - 16S rRNA gene

KW - Bacterial vaginosis

KW - Culture

KW - Hydrogen peroxide

KW - Lactobacillus

KW - Lactobacillus crispatus

KW - Lactobacillus jensenii

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U2 - 10.1186/1471-2334-12-188

DO - 10.1186/1471-2334-12-188

M3 - Article

C2 - 22889380

AN - SCOPUS:84864810775

SN - 1471-2334

VL - 12

JO - BMC Infectious Diseases

JF - BMC Infectious Diseases

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ER -

Detection of hydrogen peroxide-producing Lactobacillus species in the vagina: a comparison of culture and quantitative PCR among HIV-1 seropositive women

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

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