Methodology for a vaginal and urinary microbiome study in women with mixed urinary incontinence
Methodology for a vaginal and urinary microbiome study in women with mixed urinary incontinence
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DOI：
10.1007/s00192-016-3165-7
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发表时间：
2017-05-01
影响因子：
1.8
通讯作者：
Gantz, Marie
中科院分区：
文献类型：
--
作者：
Komesu, Yuko M.;Richter, Holly E.;Gantz, Marie
我们描述了一项旨在比较混合性尿失禁（MUI）女性和年龄相仿的无症状对照组的阴道和尿液微生物组的研究的原理和方法，本文描述了一项补充微生物组研究的方法，该研究嵌套在一项正在进行的随机对照试验中，比较了标准化围手术期行为/骨盆底运动干预加尿道中段悬吊术与单独使用尿道中段悬吊术治疗MUI。母研究中的女性在经验证的问卷中至少有“中度困扰”的紧迫性和压力性尿失禁症状（SUI），并在膀胱日记中证实了MUI。对照组无尿失禁症状。所有参与者都进行了阴道和尿液收集，用于DNA分析和常规尿培养。设计了标准化的方案，中心实验室接收样本进行随后的聚合酶链反应（PCR）扩增和细菌16 S核糖体RNA（rRNA）基因测序。将通过对阴道和尿液样本的可变区1-3和4-6进行双扩增子测序来确定细菌群落的组成，以比较患者与对照的微生物组。样本量估计确定126名MUI参与者和84名对照参与者足以检测出主要尿属的20%差异，具有80%的功效和0.05的显著性水平。标本采集于2015年1月开始，2016年4月结束。提取DNA并储存用于随后的evaluation.Methods文件共享信息的发展生殖泌尿微生物组的研究，特别是与对照人群，是很少的。我们描述了为MUI女性的新型泌尿生殖道微生物组研究开发的严格方法。
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.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.Specimen collection commenced January 2015 and finished April 2016. DNA was extracted and stored for subsequent evaluation.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.