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A paperfluidic platform to detect Neisseria gonorrhoeae in clinical samples

  • Audrey L. Horst
  • Justin M. Rosenbohm
  • Nikunja Kolluri
  • Justin Hardick
  • Charlotte A. Gaydos
  • Mario Cabodi
  • Catherine M. Klapperich
  • Jacqueline C. Linnes
Article
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ABSTRACT

Globally, the microbe Neisseria gonorrhoeae (NG) causes 106 million newly documented sexually transmitted infections each year. Once appropriately diagnosed, NG infections can be readily treated with antibiotics, but high-risk patients often do not return to the clinic for treatment if results are not provided at the point of care. A rapid, sensitive molecular diagnostic would help increase NG treatment and reduce the prevalence of this sexually transmitted disease. Here, we report on the design and development of a rapid, highly sensitive, paperfluidic device for point-of-care diagnosis of NG. The device integrates patient swab sample lysis, nucleic acid extraction, thermophilic helicase-dependent amplification (tHDA), an internal amplification control (NGIC), and visual lateral flow detection within an 80 min run time. Limits of NG detection for the NG/NGIC multiplex tHDA assay were determined within the device, and clinical performance was validated retroactively against qPCR-quantified patient samples in a proof-of-concept study. This paperfluidic diagnostic has a clinically relevant limit of detection of 500 NG cells per device with analytical sensitivity down to 10 NG cells per device. In triplicate testing of 40 total urethral and vaginal swab samples, the device had 95% overall sensitivity and 100% specificity, approaching current laboratory-based molecular NG diagnostics. This diagnostic platform could increase access to accurate NG diagnoses to those most in need.

Keywords

Neisseria gonorrhoeae point of care molecular diagnostic thermophilic helicase dependent amplification isothermal amplification internal amplification control vaginal swab urethral swab paperfluidic 
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Notes

Acknowledgements

This work was funded by the National Institute of Health National Institute of Allergy and Infectious Diseases award number R01 AI113927 to Boston University and the NIH National Institute of Biomedical and Bioengineering award number U54 EB007958 to Johns Hopkins University.

Supplementary material

10544_2018_280_MOESM1_ESM.pdf (618 kb)
ESM 1 (PDF 618 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Audrey L. Horst
    • 1
  • Justin M. Rosenbohm
    • 1
  • Nikunja Kolluri
    • 1
  • Justin Hardick
    • 2
  • Charlotte A. Gaydos
    • 2
  • Mario Cabodi
    • 1
  • Catherine M. Klapperich
    • 1
  • Jacqueline C. Linnes
    • 3
  1. 1.Department of Biomedical EngineeringBoston UniversityBostonUSA
  2. 2.School of MedicineJohns Hopkins UniversityBaltimoreUSA
  3. 3.Weldon School of Biomedical EngineeringPurdue UniversityWest LafayetteUSA

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