Stochastic confinement to detect, manipulate, and utilize molecules and organisms

Abstract

Methods of detecting organisms e.g. bacteria using stochastic confinement effects with microfluidic technologies involving plugs are provided. Signal amplification methods for the detection of molecules are also disclosed.

Description

RELATED APPLICATIONS[0001]The present patent document claims the benefit of the filing date under 35 U.S.C. §119(e) of Provisional U.S. Patent Application Ser. Nos. 60 / 962,426, filed Jul. 26, 2007, and 61 / 052,490 filed May 12, 2008, which are hereby incorporated by reference.FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]This work was supported in part by Grant No. 0526693 awarded by the National Science Foundation (NSF) CRC and under grant numbers EB01903, GM075827, and GM074961 awarded by the National Institutes of Health (NIH). The government has certain rights in the invention.BACKGROUND[0003]Bacterial infections are a major health problem, leading to more than 130,000 deaths from sepsis annually in the United States alone. (G. S. Martin, D. M. Mannino, S. Eaton and M. Moss, N. Engl. J. Med., 2003, 348, 1546-1554) These deaths are often the result of nosocomial, or hospital acquired, infections and frequently involve drug resistant strains of bacteria. (B. M. Farr, Curr. Opin....

AI Technical Summary

Benefits of technology

The patent describes methods for detecting and screening for bacteria and molecules using plugs in a microchannel. The plugs are made of a fluid that is not compatible with the carrier fluid and the bacteria or molecules produce a detectable signal when they bind to the plugs. The methods can be used for detecting bacteria in patients, screening for antibiotic activity, and detecting molecules. The technical effects of the patent include improved detection and screening for bacteria and molecules using a simple and efficient method.

Problems solved by technology

Bacterial infections are a major health problem, leading to more than 130,000 deaths from sepsis annually in the United States alone.

(B. M. Farr, Curr. Opin. Infect. Dis., 2004, 17, 317-322; G. J. Moran, A. Krishnadasan, R. J. Gorwitz, G. E. Fosheim, L. K. McDougal, R. B. Carey and D. A. Talan, N. Engl. J. Med., 2006, 355, 666-674) In addition, bacteremia, the presence of bacteria in the blood, is one of the major causes of sepsis and generally requires a minimum of a day or more to diagnose, increasing the chances of patient mortality.

(H. B. Nguyen, E. P. Rivers, F. M. Abrahamian, G. J. Moran, E. Abraham, S. Trzeciak, D. T. Huang, T. Osborn, D. Stevens and D. A. Talan, Ann. Emerg. Med., 2006, 48, 28-54) However, attempts to reduce the assay time of traditional diagnosis and characterization techniques are impeded by the necessity to incubate bacterial specimens for hours to days to increase the cell density of the sample to detectible levels.

However, these methods only provide a genetic profile of the infecting bacterial species and lack the ability to directly test the bacteria's function, such as susceptibility to particular antibiotics.

However it usually takes days to weeks to grow enough bacteria to detect them, and by the time they grow in the culture, they also grow in the patient, and the patient becomes very sick.

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