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Methodology for a vaginal and urinary microbiome study in women with mixed urinary incontinence

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Abstract

Introduction and hypothesis

We describe the rationale and methods of a study designed to compare vaginal and urinary microbiomes in women with mixed urinary incontinence (MUI) and similarly aged, asymptomatic controls.

Methods

This paper delineates the methodology of a supplementary microbiome study nested in an ongoing randomized controlled trial comparing a standardized perioperative behavioral/pelvic floor exercise intervention plus midurethral sling versus midurethral sling alone for MUI. Women in the parent study had at least “moderate bother” from urgency and stress urinary incontinence symptoms (SUI) on validated questionnaire and confirmed MUI on bladder diary. Controls had no incontinence symptoms. All participants underwent vaginal and urine collection for DNA analysis and conventional urine culture. Standardized protocols were designed, and a central lab received samples for subsequent polymerase chain reaction (PCR) amplification and sequencing of the bacterial16S ribosomal RNA (rRNA) gene. The composition of bacterial communities will be determined by dual amplicon sequencing of variable regions 1–3 and 4–6 from vaginal and urine specimens to compare the microbiome of patients with controls. Sample-size estimates determined that 126 MUI and 84 control participants were sufficient to detect a 20 % difference in predominant urinary genera, with 80 % power and 0.05 significance level.

Results

Specimen collection commenced January 2015 and finished April 2016. DNA was extracted and stored for subsequent evaluation.

Conclusions

Methods papers sharing information regarding development of genitourinary microbiome studies, particularly with control populations, are few. We describe the rigorous methodology developed for a novel urogenital microbiome study in women with MUI.

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Acknowledgments

We thank Drs. Amy Overby, the UNM CTSA T-Laboratory and Karissa Culbreath (of Tricore Laboratories) for their invaluable help with this project.

Author information

Authors and Affiliations

  1. Division of Female Pelvic Medicine and Reconstructive Surgery, Department of Obstetrics and Gynecology MSC10-5580, University of New Mexico Health Sciences Center, Albuquerque, NM, 87131-0001, USA

    Yuko M. Komesu & Rebecca G. Rogers

  2. Division of Urogynecology and Pelvic Reconstructive Surgery, Department of Obstetrics and Gynecology, University of Alabama at Birmingham, Birmingham, AL, USA

    Holly E. Richter

  3. Clinical Translational Sciences Center and Department of Pediatrics, University of New Mexico of New Mexico Health Sciences Center, Albuquerque, NM, USA

    Darrell L. Dinwiddie

  4. Division of Urogynecology and Reconstructive Pelvic Surgery, Department of Obstetrics and Gynecology, Duke University Medical Center, Durham, NC, USA

    Nazema Y. Siddiqui

  5. Division of Urogynecology and Reconstructive Pelvic Surgery, Department of Obstetrics and Gynecology, Alpert Medical School of Brown University, Providence, RI, USA

    Vivian W. Sung

  6. Division of Female Pelvic Medicine and Reconstructive Surgery, Department of Reproductive Medicine, UC San Diego Health System, San Diego, CA, USA

    Emily S. Lukacz

  7. Center for Urogynecology and Reconstructive Pelvic Surgery, Obstetrics and Gynecology and Women’s Health Institute, Cleveland Clinic, Cleveland, OH, USA

    Beri Ridgeway

  8. Division of Urogynecology and Reconstructive Pelvic Surgery, Hospital of University of Pennsylvania, Philadelphia, PA, USA

    Lily A. Arya

  9. Division of Urogynecology and Reconstructive Pelvic Surgery, Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    Halina M. Zyczynski

  10. Social, Statistical and Environmental Sciences, RTI International, Research Triangle Park, Durham, NC, USA

    Marie Gantz

Authors
  1. Yuko M. Komesu
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  2. Holly E. Richter
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  3. Darrell L. Dinwiddie
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  4. Nazema Y. Siddiqui
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  5. Vivian W. Sung
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  6. Emily S. Lukacz
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  7. Beri Ridgeway
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  8. Lily A. Arya
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  9. Halina M. Zyczynski
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  10. Rebecca G. Rogers
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  11. Marie Gantz
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Corresponding author

Correspondence to Yuko M. Komesu.

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Grants

Supported by grants from the Eunice Kennedy Shriver National Institute of Child Health and Human Development (1-U10-HD069025-01, 2-U10-HD041261-11, 2-U10-HD041267-12, 1-U10-HD069013-01, 2-U10-HD054214-06, 2-U10-HD054215-06, 1-U10-HD069010-01, 1-U10-HD069006-01, 1-U01HD069031-01) and the National Institutes of Health Office of Research on Women’s Health, and the National Center for Research Resources and the National Center for Advancing Translational Sciences of the National Institutes of Health (Grant Number ULTR001449). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Conflicts of interest

Y. Komesu: NIH 5R-01AT007171-03 (Primary Institute: NCCIH)

H. Richter: Symposia Medicus, Pelvalon, Astellas, Ferring, Statking, Society of Gynecologic Surgeons Board of Directors, UpToDate

D. Dinwiddie: None

N. Siddiqui: Medtronic

E. Lukacz: AMS/Astora, Axonics, Boston Scientific, Pfizer, Uroplasty, UpToDate

V. Sung: Society of Gynecologic Surgeons Executive Committee

B. Ridgeway: Coloplast

L. Arya: None

H. Zyczynski: American Urogynecologic Society Board of Directors, NICHD U10HD069006

R. Rogers: UpToDate, DSMB Chair TRANSFORM trial (sponsored by AMS/Astora), Mc Graw Hill, American Board of Obstetrics and Gynecology, International Urogynecology Journal

M. Gantz: None

Additional information

All authors are members of Pelvic Floor Disorders Network

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Komesu, Y.M., Richter, H.E., Dinwiddie, D.L. et al. Methodology for a vaginal and urinary microbiome study in women with mixed urinary incontinence. Int Urogynecol J 28, 711–720 (2017). https://doi.org/10.1007/s00192-016-3165-7

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  • DOI: https://doi.org/10.1007/s00192-016-3165-7

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