Performance of swabs, lavage, and diluents to quantify biomarkers of female genital tract soluble mucosal mediators

Charlene S. Dezzutti, Craig W. Hendrix, Jeanne M. Marrazzo, Zhenyu Pan, Lei Wang, Nicolette Louissaint, Sabah Kalyoussef, N. Merna Torres, Florian Hladik, Urvi Parikh, John Mellors, Sharon L. Hillier, Betsy Herold

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Background: Measurement of immune mediators and antimicrobial activity in female genital tract secretions may provide biomarkers predictive of risk for HIV-1 acquisition and surrogate markers of microbicide safety. However, optimal methods for sample collection do not exist. This study compared collection methods. Methods: Secretions were collected from 48 women (24 with bacterial vaginosis [BV]) using vaginal and endocervical Dacron and flocked swabs. Cervicovaginal lavage (CVL) was collected with 10 mL of Normosol-R (n = 20), saline (n = 14), or water (n = 14). The concentration of gluconate in Normosol-R CVL was determined to estimate the dilution factor. Cytokine and antimicrobial mediators were measured by Luminex or ELISA and corrected for protein content. Endogenous anti-HIV-1 and anti-E. coli activity were measured by TZM-bl assay or E. coli growth. Results: Higher concentrations of protein were recovered by CVL, despite a 10-fold dilution of secretions, as compared to swab eluents. After protein correction, endocervical swabs recovered the highest mediator levels regardless of BV status. Endocervical and vaginal flocked swabs recovered significantly higher levels of anti-HIV-1 and anti-E. coli activity than Dacron swabs (P<0.001). BV had a significant effect on CVL mediator recovery. Normosol-R tended to recover higher levels of most mediators among women with BV, whereas saline or water tended to recover higher levels among women without BV. Saline recovered the highest levels of anti-HIV-1 activity regardless of BV status. Conclusions: Endocervical swabs and CVL collected with saline provide the best recovery of most mediators and would be the optimal sampling method(s) for clinical trials.

Original languageEnglish (US)
Article numbere23136
JournalPLoS One
Volume6
Issue number8
DOIs
StatePublished - 2011

Fingerprint

Bacterial Vaginosis
Therapeutic Irrigation
female genitalia
Biomarkers
Human immunodeficiency virus 1
Escherichia coli
biomarkers
Polyethylene Terephthalates
Dilution
HIV-1
anti-infective agents
secretion
Recovery
Proteins
Water
Anti-Infective Agents
Assays
methodology
microbial activity
Cytokines

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Performance of swabs, lavage, and diluents to quantify biomarkers of female genital tract soluble mucosal mediators. / Dezzutti, Charlene S.; Hendrix, Craig W.; Marrazzo, Jeanne M.; Pan, Zhenyu; Wang, Lei; Louissaint, Nicolette; Kalyoussef, Sabah; Torres, N. Merna; Hladik, Florian; Parikh, Urvi; Mellors, John; Hillier, Sharon L.; Herold, Betsy.

In: PLoS One, Vol. 6, No. 8, e23136, 2011.

Research output: Contribution to journalArticle

Dezzutti, CS, Hendrix, CW, Marrazzo, JM, Pan, Z, Wang, L, Louissaint, N, Kalyoussef, S, Torres, NM, Hladik, F, Parikh, U, Mellors, J, Hillier, SL & Herold, B 2011, 'Performance of swabs, lavage, and diluents to quantify biomarkers of female genital tract soluble mucosal mediators', PLoS One, vol. 6, no. 8, e23136. https://doi.org/10.1371/journal.pone.0023136
Dezzutti, Charlene S. ; Hendrix, Craig W. ; Marrazzo, Jeanne M. ; Pan, Zhenyu ; Wang, Lei ; Louissaint, Nicolette ; Kalyoussef, Sabah ; Torres, N. Merna ; Hladik, Florian ; Parikh, Urvi ; Mellors, John ; Hillier, Sharon L. ; Herold, Betsy. / Performance of swabs, lavage, and diluents to quantify biomarkers of female genital tract soluble mucosal mediators. In: PLoS One. 2011 ; Vol. 6, No. 8.
@article{5ff2fb19ce68494f8facd87303da4316,
title = "Performance of swabs, lavage, and diluents to quantify biomarkers of female genital tract soluble mucosal mediators",
abstract = "Background: Measurement of immune mediators and antimicrobial activity in female genital tract secretions may provide biomarkers predictive of risk for HIV-1 acquisition and surrogate markers of microbicide safety. However, optimal methods for sample collection do not exist. This study compared collection methods. Methods: Secretions were collected from 48 women (24 with bacterial vaginosis [BV]) using vaginal and endocervical Dacron and flocked swabs. Cervicovaginal lavage (CVL) was collected with 10 mL of Normosol-R (n = 20), saline (n = 14), or water (n = 14). The concentration of gluconate in Normosol-R CVL was determined to estimate the dilution factor. Cytokine and antimicrobial mediators were measured by Luminex or ELISA and corrected for protein content. Endogenous anti-HIV-1 and anti-E. coli activity were measured by TZM-bl assay or E. coli growth. Results: Higher concentrations of protein were recovered by CVL, despite a 10-fold dilution of secretions, as compared to swab eluents. After protein correction, endocervical swabs recovered the highest mediator levels regardless of BV status. Endocervical and vaginal flocked swabs recovered significantly higher levels of anti-HIV-1 and anti-E. coli activity than Dacron swabs (P<0.001). BV had a significant effect on CVL mediator recovery. Normosol-R tended to recover higher levels of most mediators among women with BV, whereas saline or water tended to recover higher levels among women without BV. Saline recovered the highest levels of anti-HIV-1 activity regardless of BV status. Conclusions: Endocervical swabs and CVL collected with saline provide the best recovery of most mediators and would be the optimal sampling method(s) for clinical trials.",
author = "Dezzutti, {Charlene S.} and Hendrix, {Craig W.} and Marrazzo, {Jeanne M.} and Zhenyu Pan and Lei Wang and Nicolette Louissaint and Sabah Kalyoussef and Torres, {N. Merna} and Florian Hladik and Urvi Parikh and John Mellors and Hillier, {Sharon L.} and Betsy Herold",
year = "2011",
doi = "10.1371/journal.pone.0023136",
language = "English (US)",
volume = "6",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "8",

}

TY - JOUR

T1 - Performance of swabs, lavage, and diluents to quantify biomarkers of female genital tract soluble mucosal mediators

AU - Dezzutti, Charlene S.

AU - Hendrix, Craig W.

AU - Marrazzo, Jeanne M.

AU - Pan, Zhenyu

AU - Wang, Lei

AU - Louissaint, Nicolette

AU - Kalyoussef, Sabah

AU - Torres, N. Merna

AU - Hladik, Florian

AU - Parikh, Urvi

AU - Mellors, John

AU - Hillier, Sharon L.

AU - Herold, Betsy

PY - 2011

Y1 - 2011

N2 - Background: Measurement of immune mediators and antimicrobial activity in female genital tract secretions may provide biomarkers predictive of risk for HIV-1 acquisition and surrogate markers of microbicide safety. However, optimal methods for sample collection do not exist. This study compared collection methods. Methods: Secretions were collected from 48 women (24 with bacterial vaginosis [BV]) using vaginal and endocervical Dacron and flocked swabs. Cervicovaginal lavage (CVL) was collected with 10 mL of Normosol-R (n = 20), saline (n = 14), or water (n = 14). The concentration of gluconate in Normosol-R CVL was determined to estimate the dilution factor. Cytokine and antimicrobial mediators were measured by Luminex or ELISA and corrected for protein content. Endogenous anti-HIV-1 and anti-E. coli activity were measured by TZM-bl assay or E. coli growth. Results: Higher concentrations of protein were recovered by CVL, despite a 10-fold dilution of secretions, as compared to swab eluents. After protein correction, endocervical swabs recovered the highest mediator levels regardless of BV status. Endocervical and vaginal flocked swabs recovered significantly higher levels of anti-HIV-1 and anti-E. coli activity than Dacron swabs (P<0.001). BV had a significant effect on CVL mediator recovery. Normosol-R tended to recover higher levels of most mediators among women with BV, whereas saline or water tended to recover higher levels among women without BV. Saline recovered the highest levels of anti-HIV-1 activity regardless of BV status. Conclusions: Endocervical swabs and CVL collected with saline provide the best recovery of most mediators and would be the optimal sampling method(s) for clinical trials.

AB - Background: Measurement of immune mediators and antimicrobial activity in female genital tract secretions may provide biomarkers predictive of risk for HIV-1 acquisition and surrogate markers of microbicide safety. However, optimal methods for sample collection do not exist. This study compared collection methods. Methods: Secretions were collected from 48 women (24 with bacterial vaginosis [BV]) using vaginal and endocervical Dacron and flocked swabs. Cervicovaginal lavage (CVL) was collected with 10 mL of Normosol-R (n = 20), saline (n = 14), or water (n = 14). The concentration of gluconate in Normosol-R CVL was determined to estimate the dilution factor. Cytokine and antimicrobial mediators were measured by Luminex or ELISA and corrected for protein content. Endogenous anti-HIV-1 and anti-E. coli activity were measured by TZM-bl assay or E. coli growth. Results: Higher concentrations of protein were recovered by CVL, despite a 10-fold dilution of secretions, as compared to swab eluents. After protein correction, endocervical swabs recovered the highest mediator levels regardless of BV status. Endocervical and vaginal flocked swabs recovered significantly higher levels of anti-HIV-1 and anti-E. coli activity than Dacron swabs (P<0.001). BV had a significant effect on CVL mediator recovery. Normosol-R tended to recover higher levels of most mediators among women with BV, whereas saline or water tended to recover higher levels among women without BV. Saline recovered the highest levels of anti-HIV-1 activity regardless of BV status. Conclusions: Endocervical swabs and CVL collected with saline provide the best recovery of most mediators and would be the optimal sampling method(s) for clinical trials.

UR - http://www.scopus.com/inward/record.url?scp=80051639401&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80051639401&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0023136

DO - 10.1371/journal.pone.0023136

M3 - Article

VL - 6

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 8

M1 - e23136

ER -