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UniTaq Bio Seeking Partners for 'Universal' qPCR Tech; Developing MDR-TB Assay with Stanford


By Ben Butkus

UniTaq Bio, an early-stage company founded by a former Life Technologies R&D director, recently debuted its flagship technology — a "universal" PCR/qPCR technology that it claims will allow users to detect tens or even hundreds of nucleic acid targets in a single reaction.

The company believes its technology could simplify and reduce the cost of qPCR-based assays for a variety of research applications, and is currently seeking partnerships or licensing deals with academic labs or tool vendors in these areas.

In addition, UniTaq is seeking commercialization partners in the areas of molecular diagnostics and applied markets, and is currently collaborating with Stanford University researchers to develop multiplex assays to test for multi-drug-resistant tuberculosis

UniTaq founder Gene Spier presented his company's technology platform, also called UniTaq, during the early-stage molecular diagnostics partnering forum at last month's Molecular Medicine Tri-Conference in San Francisco.

Spier, a former senior director of R&D at Life Tech and predecessors Applied Biosystems and Celera Genomics, was at one point responsible for leading R&D and bioinformatics for TaqMan assay development, mostly for research use and some applied markets.

In an interview following the conference, Spier told PCR Insider that he founded UniTaq last year with the idea of reducing the number of pipetting steps required of scientists performing multi-well qPCR assays.

"In regular PCR experiments, you have to add sample and master mix; and in the second step you add custom TaqMan assay or SYBR Green assay to every well," Spier said. "You essentially have two pipetting steps per well. One thing I learned from my [Applied Biosystems] experience is that if you save customers pipetting, they're ready to pay a premium." Spier cited as an example the success of Life Tech's custom TaqMan Array 384-well microfluidic cards, which the company pre-loads with assay content to save customers precious time at the lab bench.

However, Spier noted, many researchers desire less-expensive options that allow them to easily create their own custom multi-well assays. To that end, he invented the UniTaq method, a two-step detection process in which multiplex PCR using primers with 5' tails "encodes" target DNA with tagging sequences, then universal dual-labeled primers detect these tags generating a 5' nuclease signal.

"With UniTaq … the assays can be preloaded in the plate, and it will be the same assay for any set of SNPs, gene expression targets, microRNAs, et cetera," Spier said. "We would pre-load the assays into 384- or 96-well plates, and then sell these pre-loaded plates, and because they're always the same assay for any set of targets, we can manufacture the plates much more cost-effectively, rather than custom-spotting a set of assays differently for each customer."

Other advantages include lower cost of goods for assays because universal labeled detection primers are "amortized over a large number of experiments;" a stronger detection signal than TaqMan, based on internal feasibility experiments;" and support of multiple dyes per well without formation of primer-dimers, something that can be a problem in SYBR Green-based assays, the company said.

UniTaq is seeking to outlicense its technology or partner with research tool companies to develop assays for a wide variety of research applications, including SNP detection, gene expression, copy number variation, microRNA detection, somatic mutation detection, and methylation assessment.

Spier said that UniTaq's technology brings similar multiplexing advantages to applied market applications, such as water, food, and environmental testing; as well as to molecular diagnostics. Even though UniTaq assays are qualitative, and only indicate whether a target nucleic acid is present — as opposed to which target is present — in a background of non-targets, they would still be ideal in situations where the tester is required to perform a large amount of screens but is really only looking for the presence of a specific target.

For instance, in the example of multi-drug-resistant tuberculosis, "there can be multiple mutations … and if any of them is present, then the drug rifampicin will not work," Spier said.

"With TaqMan assays, if you were looking to detect one mutation, you would, generally speaking, need … one well for each of the mutations," he added. "But with UniTaq, you can do it in a single well. You won't know which of the nine mutations is present, but if any one of them is present you get a signal. And from a diagnostic perspective, that is sufficient to know not to take this particular drug and instead take a second-line treatment that is not resistant to the drug."

The same holds true for food, water, and environmental testing, he said.

"You would need to know if an E. coli plasmid is present … but most of the tests are negative," he said. "When something shows up you do a more detailed test — sometimes you sequence. In the relatively rare event that something is present and you really need to know what it is, you can do a follow-up test. But this is much rarer than the screening itself, which happens in almost every batch of food processing."

UniTaq seeks collaborators on both the applied markets and diagnostics front, but it is also working with the Gary Schoolnick laboratory at Stanford to develop its own diagnostic development project in the aforementioned area of MDR-TB. More specifically, the group is developing an assay that would be able to test for resistance to rifampicin, isoniazid, and ethambutol in a single qPCR well.

"This is very early-stage, so we are also applying for grants, and in fact have two pending grant applications," Spier said. "We haven't even established our own lab. One way for us to raise some money is to license this in a field where we wouldn't develop [assays] anyway."

Another way to raise money, of course, is through venture capitalists, and Spier has begun discussions on that front, as well. "Most of these investors also have portfolio companies that could be using this technology, so that becomes a little more interesting for them," he said.

UniTaq has filed a PCT patent and provisional patent with the US Patent and Trademark Office, and Spier said he is "very confident" that they will be awarded. "The combination of novelty and usefulness is there. Maybe some claims could be modified." UniTaq's primer design has "some similarity, topology-wise," to existing patented PCR primers, but those are generally target-specific primers while UniTaq is universal, he added.

"Whether this is enough difference for freedom to operate is very hard to say," he said. "You can find lawyers to argue either way."

Have topics you'd like to see covered in PCR Insider? Contact the editor at bbutkus [at] genomeweb [.] com.

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