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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
M. Alary, C. Gbenafa-Agossa, G. Aina, et al., Evaluation of a rapid point-of-care test for the detection of gonococcal infection among female sex workers in Benin. Sex. Transm. Infect. 82, 29–32 (2006)
J.E. Aledort, A. Ronald, M.E. Rafael, et al., Reducing the burden of sexually transmitted infections in resource-limited settings: the role of improved diagnostics. Nature 444, 59–72 (2006)
A. Bourgeois, D. Henzel, G. Malonga-Mouelet, et al., Clinical algorithms for the screening of pregnant women for STDs in Libreville, Gabon: which alternatives? Sex. Transm. Infect. 74, 35–39 (1998)
R.M. Brotman, Vaginal microbiome and sexually transmitted infections: an epidemiologic perspective. J. Clin. Investig. 121, 4610–4617 (2011)
Cortez Diagnostics, Inc. OneStep RapiDip Gonorrhea InstaTest Package Insert. (Cortez Diagnostics, Inc., 2006). http://www.rapidtest.com/Gonorrhea_176512.pdf. Accessed 2 Dec 2016
J.P. Dillard, Genetic Manipulation of Neisseria gonorrhoeae. Curr. Protoc. Microbiol. 23, 4A.1.1–4A.1.24 (2011)
D.T. Fleming, J.N. Wasserheit, From epidemiological synergy to public health policy and practice: the contribution of other sexually transmitted diseases to sexual transmission of HIV infection. Sex. Transm. Infect. 75, 3–17 (1999)
C.A. Gaydos, B.V.D. Pol, M. Jett-Goheen, et al., Performance of the Cepheid CT/NG Xpert Rapid PCR Test for detection of Chlamydia trachomatis and Neisseria gonorrhoeae. J. Clin. Microbiol. 51, 1666–1672 (2013)
L. Gervais, E. Delamarche, Toward one-step point-of-care immunodiagnostics using capillary-driven microfluidics and PDMS substrates. Lab. Chip. 9, 3330 (2009)
L. Greer, G.D. Wendel, Rapid diagnostic methods in sexually transmitted infections. Infect. Dis. Clin. N. Am. 22, 601–617 (2008)
R.J. Guy, L.M. Cuaser, J.D. Klausner, et al., Performance and operational characteristics of point-of-care tests for the diagnosis of urogenital gonococcal infections. Sex. Transm. Dis. 93, S16–S21 (2017)
J. Huppert, E. Hesse, C.A. Gaydos, What is the point? How point-of-care sexually transmitted infection tests can impact infected patients. Point Care 9, 36–46 (2010)
R.E. Johnson, W.J. Newhall, J.R. Papp, et al., Screening tests to detect Chlamydia trachomatis and Neisseria gonorrhoeae infections–2002. MMWR Recomm. Rep 51, 1–38 (2002)
M.D. Kulinski, M. Mahalanabis, S. Gillers, et al., Sample preparation module for bacterial lysis and isolation of DNA from human urine. Biomed. Microdevices 11, 671–678 (2009)
L.K. Lafleur, J.D. Bishop, E.K. Heiniger, et al., A rapid, instrument-free, sample-to-result nucleic acid amplification test. Lab Chip 16, 3777–3787 (2016)
J.C. Linnes, A. Fan, N.M. Rodriguez, et al., Paper-based molecular diagnostic for Chlamydia trachomatis. RSC Adv. 4, 42245–42251 (2014)
J.C. Linnes, N.M. Rodriguez, L. Liu, et al., Polyethersulfone improves isothermal nucleic acid amplification compared to current paper-based diagnostics. Biomed. Microdevices 18, 30 (2016)
T.L. Lowe, S.J. Kraus, Quantitation of Neisseria gonorrhoeae from women with gonorrhea. J. Infect. Dis. 133, 621–626 (1976)
J.R. Papp, J. Schachter, CA Gaydos, et al., Recommendations for the laboratory-based detection of Chlamydia trachomatis and Neisseria gonorrhoeae–2014. MMWR Recomm. Rep. 63, 1–19 (2014)
L.B. Price, C.M. Liu, K.E. Johnson, et al., The effects of circumcision on the penis microbiome. PLoS One 5, 1–12 (2010)
D. Priest, J.J. Ong, E.P.F. Chow, et al., Neisseria gonorrhoeae DNA bacterial load in men with symptomatic and asymptomatic gonococcal urethritis. Sex. Transm. Infect. 93(7), 478–481 (2017)
N.M. Rodriguez, W.S. Wong, L. Liu, et al., A fully integrated paperfluidic molecular diagnostic chip for the extraction, amplification, and detection of nucleic acids from clinical samples. Lab Chip 16, 753–763 (2016)
S. Roy, C. Yue, Z. Wang, L. Anand, Thermal bonding of microfluidic devices: Factors that affect interfacial strength of similar and dissimilar cyclic olefin copolymers. Sensors Actuators B Chem. 161, 1067–1073 (2012)
M. Toby, P. Saunders, M. Cole, et al., Prevalence of porA pseudogene deletion among Neisseria gonorrhoeae isolates referred to the UK's Gonococcal Resistance to Antimicrobials Surveillance Program. Sex. Health 14(4), 392–393 (2017)
I. Toskin, M. Murtagh, R.W. Peeling, et al., Advancing prevention of sexually transmitted infections through point-of-care testing: target product profiles and landscape analysis. Sex. Transm. Dis. 93, S69–S80 (2017)
U.S. Food and Drug Administration. Draft Guidance for Industry and Food and Drug Administration Staff Establishing the Performance Characteristics of In Vitro Diagnostic Devices for Chlamydia trachomatis and/or Neisseria gonorrhoeae: Screening and Diagnostic Testing. U.S. Federal Register. 76(91), 19 (2011)
M. Unemo, O. Norlen, H. Fredlund, The porA pseudogene of Neisseria gonorrhoeae - low level of genetic polymorphism and a few, mainly identical, inactivating mutations. APMIS 113, 410–419 (2005)
J.N. Wasserheit, Epidemiological synergy. Interrelationships between human immunodeficiency virus infection and other sexually transmitted diseases. Sex. Transm. Dis. 19, 61–77 (1992)
World Health Organization, Global strategy for the prevention and control of sexually transmitted infections : 2006 - 2015 : breaking the chain of transmission. (WHO Press, Geneva, Switzerland 2007) p. 26
World Health Organization, Global incidence and prevalence of selected curable sexually transmitted infections – 2008. (WHO Press, Geneva, Switzerland, 2012) p. 2
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.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
ESM 1
(PDF 618 kb)
Rights and permissions
About this article
Cite this article
Horst, A.L., Rosenbohm, J.M., Kolluri, N. et al. A paperfluidic platform to detect Neisseria gonorrhoeae in clinical samples. Biomed Microdevices 20, 35 (2018). https://doi.org/10.1007/s10544-018-0280-x
Published:
DOI: https://doi.org/10.1007/s10544-018-0280-x