Source: HJ Science & Technology, Inc. submitted to
LOW-COST, HANDHELD MICROFLUIDIC DEVICE FOR DETECTION AND MONITORING OF BREVETOXINS IN MARINE ENVIRONMENT
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
Reporting Frequency
Annual
Accession No.
0233694
Grant No.
2013-33610-20818
Cumulative Award Amt.
$99,863.00
Proposal No.
2013-00318
Multistate No.
(N/A)
Project Start Date
Jul 1, 2013
Project End Date
Feb 28, 2015
Grant Year
2013
Program Code
[8.7]- Aquaculture
Project Director
Jiao, H.
Recipient Organization
HJ Science & Technology, Inc.
187 Saratoga Avenue
Santa Clara,CA 95050
Performing Department
(N/A)
Non Technical Summary
During harmful algal blooms (HAB), brevetoxins are concentrated enough to cause a combination of gastro-intestinal and neurological symptoms as well as other adverse human health effects. Neurotoxic Shellfish Poisoning is caused by consumption of shellfish contaminated with brevetoxins during HAB. Rapid and real-time methods for brevetoxin detection are necessary to protect the public health and ensure food safety. Current measurement and detection methods rely on having to take the samples from the field to the laboratory for analysis. This is often a time consuming and costly process. More importantly, the lack of real-time data hampers proper and timely decision making. The proposed handheld platform is capable of rapid and real-time detections of brevetoxins with the selectivity and sensitivity that can only be achieved today with laboratory instruments. The proposed technology will therefore greatly benefit the shellfish industry and the aquaculture industry in general. In addition to brevetoxins, the same technology has clear potential to be subsequently developed to permit the diagnostics of toxins produced by a wide range of other pathogens. In particular, the same microfluidic instrumentation could be readily adapted to monitoring other water-based toxins including additional marine HAB toxins associated with fish and shellfish contamination as well as cyanobacteria toxins.
Animal Health Component
100%
Research Effort Categories
Basic
(N/A)
Applied
100%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
4025310202075%
1334099110325%
Goals / Objectives
The goal of this SBIR research project is to demonstrate the feasibility of an integrated microfluidic technology capable of rapid and real-time detections and monitoring of brevetoxins, neurotoxins produced by the dinoflagellate, Karenia brevis, in the Gulf of Mexico and other marine environment. The proposed technology is the result of the infusion of our proprietary automated on-chip microfluidic assay technology with the built-in features of smart phones in a miniaturized format. As such, it is inexpensive, easy to use and has a handheld platform. Our Phase I objective is to adapt the commercially established immunoassay protocols to our integrated microfluidic platform with a disposable microfluidic cartridge. If successful, we will be well positioned to address the needs of real-time detection of brevetoxins, and other algal biotoxins as handheld, user-friendly detectors are in great demand for protecting public health and coastal community economies. To this end, the proposed research effort will satisfy one of the five primary USDA NIFA Societal Challenge Areas: Food Safety. The proposed technology will help reduce the incidence of food-borne illnesses and death through a safe food supply and improve the ability to detect food-borne toxins and pathogens.
Project Methods
The proposed technology is the result of the infusion of our proprietary automated on-chip microfluidic assay technology with the built-in features of smart phones in a miniaturized format. In particular, we will transfer existing commercial enzyme linked immunosorbent assay protocols to the integrated microfluidic format, enabling us to develop a rapid, fully-automated immunoassay platform. As smart phones are becoming ubiquitous, they provide a robust, reconfigurable, and inexpensive platform for interfacing to the microfluidic device. By leveraging these built-in features of commercial smart phones such as wireless communication, information storage, optical imaging and signal processing, the design of the handheld device is greatly simplified and the manufacturing cost is significantly reduced. As a result, the proposed technology has a number of advantages including reducing the overall footprint, cost, and the power consumption of the device, offering more flexible information management and better overall device performance. The Phase I technical work plan includes 1) fabrication of the disposable microfluidic chip, 2) demonstrating on-chip automated assay preparation for brevetoxins, and 3) developing the instrumentation for interfacing built-in features of the smart phone to the microfluidic device. During the subsequent Phase II effort, we will build a handheld instrument with digital readout and wireless data transfer feature capable of rapid and real-time detections of brevetoxins and other water-based toxins with the selectivity and sensitivity that can only be achieved today with laboratory instruments.

Progress 07/01/13 to 02/28/15

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? In this successful SBIR Phase I effort, HJS&T has established the technical feasibility of the microfluidic automation device to perform on-site and rapid analysis for algal toxins that cause shellfish poisoning in the marine environment The Phase I feasibility study employs saxitoxin (STX) and okadaic acid (OA) as examples, but the results can be readily extended to other marine algal toxins. Specifically in Phase I, we have adapted direct competitive ELISA procedures from a commercial kit (Abraxis, Warminster, PA) to our microfluidic automation platform for both STX and OA by using both standards and real samples from natural marine environment. The analysis from marine samples includes microfluidic automation of sample preparations such as cell lysis and, as a result, measures both extra- and intra-cellular toxins concentrations. The microfluidic automation results agree with manually prepared commercial kits. As such, we have achieved the overall Phase I goal of demonstrating microfluidic automation of direct competitive ELISA for both STX and OA by accomplishing all five of the following technical objectives: We have successfully designed and fabricated several microfluidic devices capable of performing automated ELISA for up to 5 standards and/or samples with duplicates simultaneously. We have designed and built the necessary optical detection instrumentation. We have successfully adapted ELISA protocol to our microfluidic automation platform for STX by a) generating a standard curve of 0.05, 0.10, 0.20, and 0.40 ppb of seawater STX standards that is in agreement with the commercial ELISA kit in terms of limit of detection (LOD), R2 fit, and detection resolution, and b) analyzing seawater samples containing various STX producing Alexandrium strains from Professor Kudela's laboratory in UC Santa Cruz and quantifying the combined intra and extra-cellular STX concentrations that are in agreement with the commercial ELISA kit. We have successfully adapted ELISA protocol to our microfluidic automation platform for OA by a) generating a standard curve of 0.2, 1.0, and 5.0 ppb of seawater OA standards that is in excellent agreement with the commercial ELISA kit in terms of LOD, R2 fit, and detection resolution, and b) analyzing local seawater and finding no measurable amount of interference agents in the sample (< 0.2 ppb, the LOD), consistent with the commercial ELISA kit. We have designed a Phase II prototype microfluidic device with expanded sample automation capabilities. In addition to the five original Phase I objectives, we have successfully performed microfluidic automation of real sample preparation: chemical cell lysis. The results are in excellent agreement with the commercial cell lysis kit. As such, the Phase I result has surpassed the goal of the original Phase I proposal by successful demonstration of microfluidic automation of both sample preparation and ELISA analysis.

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