Research for faster Polymerase Chain Reaction technology

Imagine being able to diagnose a viral strain at the site of an outbreak or identifying a deadly pathogen on the battlefield.

University of Nebraska-Lincoln professors Hendrik Viljoen (chemical engineering) and George Gogos (mechanical engineering) have been awarded a $1.44 million grant over five years from the National Institutes of Health to further their development of faster Polymerase Chain Reaction technology. PCR is a technique for amplifying DNA for diagnostic purposes wherein a sample of target DNA is replicated many times over to enable gene sequencing and identification.

"The key to our technology is that we're able to amplify DNA in five to 10 minutes," Viljoen said. "Our device is also rugged, transportable and reliable. The rapid PCR uniquely positions us to pursue two other technologies. We can measure the PCR kinetics and the results are included in intelligent software, which will be part of the next generation of PCR thermocyclers. We have also started a program in assembly PCR. Short pieces of DNA are assembled, under conditions to minimize mutations, into larger structures."

To enable this rapid amplification, Viljoen and Gogos have integrated a novel device to produce the heating and cooling gases necessary for the thermocycling of the DNA.

"This device makes our technology amenable to field usage," Viljoen said. "And it can handle volumes from 5 microliters to 40 microliters with outstanding sensitivity, while producing a high yield."

The grant will be used to add optical detection, establish protocols for reverse transcription and the quantification of PCR, as well as to collect kinetic data of various polymerase enzymes to create mathematical models. The researchers have partnered with Michael Nelson, president of the Lincoln biotechnology firm Megabase Research Products. The firm provides the required biochemistry expertise.

"This is an interdisciplinary project. Putting together a group of chemical engineers, mechanical engineers and biochemists has been very critical to the success of the project," Gogos said. "We have built a device with vast medical and non-medical applications which at the same time is a scientific instrument for basic studies."

The implications for this innovative technology are many. "We hope to form cross-disciplinary teams of researchers to look at parallel applications of this technology," said Dipanjan Nag, a technology development associate with the UNL Office of Technology Development, which provided the researchers with an additional $75,000 grant for further development of the technology.

"This NIH grant demonstrates that we are at the cutting edge of research," Nag said. "And it has tremendous potential in biosecurity- and bioterrorism-related research."