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.
| Original language | English (US) |
|---|---|
| Article number | 35 |
| Journal | Biomedical Microdevices |
| Volume | 20 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jun 1 2018 |
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Keywords
- internal amplification control
- isothermal amplification
- molecular diagnostic
- Neisseria gonorrhoeae
- paperfluidic
- point of care
- thermophilic helicase dependent amplification
- urethral swab
- vaginal swab
ASJC Scopus subject areas
- Biomedical Engineering
- Molecular Biology
Cite this
A paperfluidic platform to detect Neisseria gonorrhoeae in clinical samples. / Horst, Audrey L.; Rosenbohm, Justin M.; Kolluri, Nikunja; Hardick, Justin; Gaydos, Charlotte A; Cabodi, Mario; Klapperich, Catherine M.; Linnes, Jacqueline C.
In: Biomedical Microdevices, Vol. 20, No. 2, 35, 01.06.2018.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A paperfluidic platform to detect Neisseria gonorrhoeae in clinical samples
AU - Horst, Audrey L.
AU - Rosenbohm, Justin M.
AU - Kolluri, Nikunja
AU - Hardick, Justin
AU - Gaydos, Charlotte A
AU - Cabodi, Mario
AU - Klapperich, Catherine M.
AU - Linnes, Jacqueline C.
PY - 2018/6/1
Y1 - 2018/6/1
N2 - 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.
AB - 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.
KW - internal amplification control
KW - isothermal amplification
KW - molecular diagnostic
KW - Neisseria gonorrhoeae
KW - paperfluidic
KW - point of care
KW - thermophilic helicase dependent amplification
KW - urethral swab
KW - vaginal swab
UR - http://www.scopus.com/inward/record.url?scp=85045283796&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85045283796&partnerID=8YFLogxK
U2 - 10.1007/s10544-018-0280-x
DO - 10.1007/s10544-018-0280-x
M3 - Article
VL - 20
JO - Biomedical Microdevices
JF - Biomedical Microdevices
SN - 1387-2176
IS - 2
M1 - 35
ER -