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Rapid electrostatic DNA enrichment for sensitive detection of Trichomonas vaginalis in clinical urinary samples

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Abstract

Estimated to be the most common non-viral sexually transmitted infection globally, Trichomonas vaginalis (TV) can lead to pelvic inflammatory disease, pregnancy complications, and increased risk of acquiring and transmitting HIV. Once diagnosed, TV infection can be treated with oral antibiotics; however, infected individuals are often asymptomatic and do not seek treatment. The WHO and others have identified a need for point-of-care tests to expand access to TV testing and screening; ideal test characteristics include high sensitivity and specificity and the ability to use urine as a sample type, rather than invasively collected swab samples. Here, we report on a proof-of-concept prototype for rapid, electrostatic enrichment of DNA from urine samples and demonstrate the use of large volumes of urine to increase sensitivity of downstream nucleic acid amplification testing. We developed an internally controlled thermophilic helicase-dependent amplification (tHDA) assay with lateral flow immunoassay readout and demonstrate that this tHDA assay can be performed directly on our DNA capture filter. We validated our method using clinical urine samples with qPCR-quantified TV loads. Using 62 clinical urine samples and a simple sample processing device, our tHDA assay displayed 96.6% sensitivity and 100% specificity. Our analytical limit of detection was found to be approximately 7 genomic equivalents of TV DNA per mL of sample when 1 mL of sample was tested, comparable to existing isothermal tests for TV. Using large-volume simulated samples (40 mL of buffered urine with spiked-in TV DNA), we also demonstrated that sensitivity could be improved 28-fold to 0.25 genomic equivalents of TV DNA per mL, with a sample processing time of only 2 minutes.

Graphical abstract: Rapid electrostatic DNA enrichment for sensitive detection of Trichomonas vaginalis in clinical urinary samples

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Supplementary files

Article information


Submitted
18 Nov 2019
Accepted
03 Feb 2020
First published
05 Feb 2020

Anal. Methods, 2020, Advance Article
Article type
Paper

Rapid electrostatic DNA enrichment for sensitive detection of Trichomonas vaginalis in clinical urinary samples

J. M. Rosenbohm, J. M. Robson, R. Singh, R. Lee, J. Y. Zhang, C. M. Klapperich, N. R. Pollock and M. Cabodi, Anal. Methods, 2020, Advance Article , DOI: 10.1039/C9AY02478F

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