Final Report: Handheld microfluidic device for cyanobacteria toxin detection and monitoring

EPA Contract Number: EPD15007
Title: Handheld microfluidic device for cyanobacteria toxin detection and monitoring
Investigators: Jiao, Hong
Small Business: HJ Science & Technology Inc.
EPA Contact: Manager, SBIR Program
Phase: II
Project Period: November 1, 2014 through October 31, 2016 (Extended to October 31, 2017)
Project Amount: $299,954
RFA: Small Business Innovation Research (SBIR) - Phase II (2014) Recipients Lists
Research Category: Small Business Innovation Research (SBIR) , SBIR - Water

Description:

In this EPA Phase II SBIR project, HJ Science & Technology, Inc. sought to develop a portable microfluidic technology capable of performing rapid and on-site detection and identification of microcystins and other toxins produced by freshwater cyanobacteria (cyanotoxins). During blooms, the cyanotoxin concentrations are high enough to cause damage to liver or damage to nerve axons and synapses. As a result, the United State EPA recommends the level of microcystins, cylindrospermopsins, and saxitoxins in drinking water not to exceed 1 part-per-billion (ppb). Currently, cyanotoxin levels are monitored by collecting the samples in the field and bringing them back to the laboratory for analysis. Compared with laboratory based methods, our portable on-site platform offers several important advantages including reduction in time and reagent consumption, and real-time data for better and more timely decision making. During the Phase II effort, we have designed, built, and tested a handheld prototype capable of performing on-site detection of microcystin. The heart of our innovation is a novel microfluidic device. As such, the majority of the Phase II effort entails developing and refining the microfluidic device as well as miniaturizing the supporting hardware. The handheld Phase II prototype is inexpensive, easy to use, and designed to perform rapid cyanotoxin detections with sensitivity and specificity that are only currently achievable with laboratory based instruments. We have performed field testing with our Phase II prototype at a local lake. Finally, we have patented our microfluidic technology and have since licensed it along with the related Intellectual Property (IP) and "know-how" to Protein Fluidics, Inc. (PFI) for commercialization as a part of the Phase II Commercialization Option. PFI has secured the necessary funds from private investors and has begun manufacturing for the commercial market. The key target audiences include resource managers, public health officials, and aquaculture facilities.

Summary/Accomplishments (Outputs/Outcomes):

The main Phase II SBIR effort entails developing and refining microfluidic device design, resulting in 4 generations of microfluidic devices for performing rapid and on-site cyanotoxin detection. The first generation microfluidic device is based on our Phase I microvalve technology. We have demonstrated the capability of the first generation 2 microfluidic technology to detect microcystin with comparable results as the conventional laboratory based commercial ELISA kit. However, the approach has serious commercial limitations in terms of complex manufacture process and the associated costs. In response to these shortcomings, we have developed a second generation microfluidic device by inventing a new technology: valve-less fluidic switching (VLFS). We have filed patent application, and the patent has been issued (US Patent Number 9,733,239: "Reconfigurable microfluidic systems: Scalable, multiplexed immunoassays"). The VLFS technology addresses the limitations of our old microvalve based microfluidic technology by eliminating on-chip valves or other moving parts, thereby eliminating the costly and labor-intensive microvalve fabrication processes. We have employed the VLFS based second generation microfluidic device to perform automated detection of microcystin with comparable results as commercial ELISA kit, but with a shorter overall assay time (40 minutes vs. 90 minutes). However, the supporting hardware instrumentation is too bulky and expensive for field deployable applications. As a result, we developed a third generation microfluidic device, where we exploit the recent technological advances in the smartphone by using the smartphone camera as the detector such that the overall footprint and the cost of the prototype instrument are significantly reduced. Finally, we developed a fourth generation microfluidic device, where the novel microfluidic channel design significantly reduces the overall assay time compared to the previous generation microfluidic devices: from 40 minutes to less than 10 minutes. The fourth generation microfluidic device has comparable performance as the commercial ELISA kit to detect microcystin and other cyanotoxins both in terms of detection sensitivity and reproducibility. We have incorporated the fourth generation microfluidic device into our handheld Phase II prototype. Finally, we have performed field testing with the handheld Phase II prototype at a local lake.

Conclusions:

In this EPA Phase II SBIR project, HJ Science & Technology, Inc. has successfully developed a portable microfluidic technology capable of performing rapid and on-site detection and identification of microcystins and other cyanotoxins. We have incorporated the microfluidic technology into the handheld Phase II prototype, which is inexpensive, easy to use, and designed to perform rapid and on-site cyanotoxin detections with sensitivity and specificity that are only currently achievable with laboratory based instruments. We have employed the handheld Phase II prototype to perform field testing at a local lake. Compared with laboratory based methods, our portable on-site technology offers several important advantages including reduction in time and reagent consumption, and real-time data for better and more timely decision making. During the course of the Phase II SBIR project, we have also patented our microfluidic technology and have since licensed it along with the related Intellectual Property (IP) and "know-how" to Protein Fluidics, Inc. (PFI) for commercialization as a part of the Phase II Commercialization Option. PFI has secured the necessary funds from private investors and has begun manufacturing for the commercial market.

Supplemental Keywords:

Microfluidics, microcystin, harmful algal bloom


SBIR Phase I:

Handheld Microfluidic Device for Cyanobacteria Toxin Detection and Monitoring  | Final Report