Title: Integrated LAMP and immunoassay platform for diarrheal disease detection

Abstract

The challenges of diagnosing infectious disease, especially in the developing world, and the shortcomings of available instrumentation have exposed the need for portable, easy-to-use diagnostic tools capable of detecting the wide range of causative microbes while operating in low resource settings. Here, we present a centrifugal microfluidic platform that combines ultrasensitive immunoassay and isothermal amplification-based screening for the orthogonal detection of both protein and nucleic acid targets at the point-of-care. A disposable disc with automatic aliquoting inlets is paired with a non-contact heating system and precise rotary control system to yield an easy-to-use, field-deployable platform with versatile screening capabilities. The detection of three enterotoxins (cholera toxin, Staphylococcal enterotoxin B, and Shiga-like toxin 1) and three enteric bacteria (C. jejuni, E. coli, and S. typhimurium) were performed independently and shown to be highly sensitive (limit of detection = 1.35–5.50 ng/mL for immunoassays and 1–30 cells for isothermal amplification), highly exclusive in the presence of non-specific targets, and capable of handling a complex sample matrix like stool. The full panel of toxins and bacteria were reliably detected simultaneously on a single disc at clinically relevant sample concentrations in less than an hour. The ability of our technology to detect multiple analyte typesmore » in parallel at the point-of-care can serve a variety of needs, from routine patient care to outbreak triage, in a variety of settings to reduce disease impact and expedite effective treatment.« less

Authors:

Phaneuf, Christopher R.; Mangadu, Betty; Tran, Huu M.; Light, Yooli K.; Sinha, Anchal; Charbonier, Frank W.; Eckles, Tyler P.; Singh, Anup K.; Koh, Chung-Yan

Publication Date:

Thu Nov 01 00:00:00 EDT 2018

Research Org.:

Sandia National Lab. (SNL-CA), Livermore, CA (United States)

Sponsoring Org.:

USDOE National Nuclear Security Administration (NNSA); US Dept. of Homeland Security (DHS); National Institutes of Health (NIH)

Contributing Org.:

US Dept. of Agriculture (USDA) Agricultural Research Service (ARS)

OSTI Identifier:

1617389

Alternate Identifier(s):

OSTI ID: 1472252

Report Number(s):

SAND2018-9227J
Journal ID: ISSN 0956-5663; S0956566318305943; PII: S0956566318305943

Grant/Contract Number:  

R01AI098853; NA0003525; HSHQPM-13-X-00222; AC04-94AL85000

Resource Type:

Published Article

Journal Name:

Biosensors and Bioelectronics

Additional Journal Information:

Journal Name: Biosensors and Bioelectronics Journal Volume: 120 Journal Issue: C; Journal ID: ISSN 0956-5663

Publisher:

Elsevier

Country of Publication:

Netherlands

Language:

English

Subject:

47 OTHER INSTRUMENTATION; 59 BASIC BIOLOGICAL SCIENCES; Microfluidics; Centrifugal; Point-of-care; Immunoassay; Isothermal amplification; LAMP; Enteric pathogens