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

Dezzutti, CS and Hendrix, CW and Marrazzo, JM and Pan, Z and Wang, L and Louissaint, N and Kalyoussef, S and Torres, NM and Hladik, F and Parikh, U and Mellors, J and Hillier, SL and Herold, BC (2011) Performance of swabs, lavage, and diluents to quantify biomarkers of female genital tract soluble mucosal mediators. PLoS ONE, 6 (8).

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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. © 2011 Dezzutti et al.


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Item Type: Article
Status: Published
CreatorsEmailPitt UsernameORCID
Dezzutti, CScsd13@pitt.eduCSD13
Hendrix, CW
Marrazzo, JM
Pan, Z
Wang, L
Louissaint, N
Kalyoussef, S
Torres, NM
Hladik, F
Parikh, Uump3@pitt.eduUMP3
Mellors, Jjwm1@pitt.eduJWM1
Hillier, SLslh6@pitt.eduSLH6
Herold, BC
ContributionContributors NameEmailPitt UsernameORCID
Date: 18 August 2011
Date Type: Publication
Journal or Publication Title: PLoS ONE
Volume: 6
Number: 8
DOI or Unique Handle: 10.1371/journal.pone.0023136
Schools and Programs: School of Medicine > Obstetrics, Gynecology, and Reproductive Sciences
Refereed: Yes
MeSH Headings: Adult; Analysis of Variance; Biological Markers--analysis; Cervix Uteri--metabolism; Cervix Uteri--microbiology; Cytokines--analysis; Cytokines--immunology; Female; Genitalia, Female--metabolism; HIV Infections--diagnosis; HIV Infections--immunology; HIV Infections--prevention & control; HIV-1--immunology; Humans; Interleukin-8--analysis; Interleukin-8--immunology; Mucous Membrane--metabolism; Risk Factors; Secretory Leukocyte Peptidase Inhibitor--analysis; Secretory Leukocyte Peptidase Inhibitor--immunology; Solubility; Specimen Handling--methods; Therapeutic Irrigation--methods; Vagina--metabolism; Vagina--microbiology; Vaginal Smears--methods; Vaginosis, Bacterial--immunology; Vaginosis, Bacterial--microbiology; Young Adult; alpha-Defensins--analysis; alpha-Defensins--immunology
Other ID: NLM PMC3155537
PubMed Central ID: PMC3155537
PubMed ID: 21858008
Date Deposited: 05 Sep 2012 19:02
Last Modified: 27 Feb 2019 10:55


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