This project will demonstrate a powerful new strategy for diagnosing infectious diseases in minimally equipped laboratories or at point of care (POC). Developed for biodefense and similar low-complexity testing purposes, the RAMP(R) antigen detection system (Response Biomedical Corporation, Vancouver, BC, Canada) will be adapted for POC diagnosis of tuberculosis (TB). The RAMP system has been extensively evaluated by independent testers, and its high sensitivity and outstanding robustness have been demonstrated. Its sensitivity will be enhanced by integration with a novel electrohydrodynamic microtip system designed to selectively concentrate Mycobacterium tuberculosis (MTB) cells in sputum samples prior to RAMP analysis. The microtip system was developed and validated by NanoFacture, Inc., in collaboration with the Nanomanufacturing Laboratory at the University of Washington. The integrated microtip-RAMP test will detect MTB cells with a lower limit of detection of <100 cfu per mL of sputum, comparable to PCR in a 20 min total test time.
Specific aims of the project are: 1) To develop a RAMP test for MTB cells in sputum;2) to develop sample processing procedures that utilize the microtip system to concentrate MTB cells from human sputum;and 3) to evaluate the performance of the microtip-RAMP system in a field trial in Dhaka, Bangladesh. Targeted pre-clinical performance for the 20-min microtip-RAMP test will have sensitivity and specificity of >90% relative to culture. In addition, the hypothesis will be tested that microtip-RAMP is significantly more sensitive than AFB smear microscopy, currently the world's most widely used rapid test to detect MTB cells in sputum.

Public Health Relevance

The project will develop and evaluate a novel tuberculosis diagnostic test that is sensitive, specific, rapid, inexpensive, and amenable to use at point-of-care or in minimally equipped laboratories. The microtip-RAMP system will be a versatile platform that can be applied to numerous other infectious diseases, individually or in multiplex fashion.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI093418-02
Application #
8321433
Study Section
Special Emphasis Panel (ZAI1-LG-M (J3))
Program Officer
Jacobs, Gail G
Project Start
2011-08-17
Project End
2012-12-18
Budget Start
2012-08-01
Budget End
2012-12-18
Support Year
2
Fiscal Year
2012
Total Cost
$82,218
Indirect Cost
$12,817
Name
Seattle Biomedical Research Institute
Department
Type
DUNS #
070967955
City
Seattle
State
WA
Country
United States
Zip Code
98109
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Inoue, Shinnosuke; Becker, Annie L; Kim, Jong-Hoon et al. (2014) Semi-automated, occupationally safe immunofluorescence microtip sensor for rapid detection of Mycobacterium cells in sputum. PLoS One 9:e86018
Kim, Jong-Hoon; Hiraiwa, Morgan; Lee, Hyun-Boo et al. (2013) Electrolyte-free Amperometric Immunosensor using a Dendritic Nanotip. RSC Adv 3:4281-4287
Chen, Wanyu; Kim, Jong-Hoon; Zhang, Di et al. (2013) Microfluidic one-step synthesis of alginate microspheres immobilized with antibodies. J R Soc Interface 10:20130566
Weigel, Kris M; Jones, Kelly L; Do, Julie S et al. (2013) Molecular viability testing of bacterial pathogens from a complex human sample matrix. PLoS One 8:e54886
Shu, Zhiquan; Weigel, Kris M; Soelberg, Scott D et al. (2012) Cryopreservation of Mycobacterium tuberculosis complex cells. J Clin Microbiol 50:3575-80