Our expertise in enzyme technology can help your idea become a commercial reality
Enteroviral (EV) meningitis is very common among young children; however, it is very difficult to differentiate from bacterial meningitis due to similar clinical presentations. A robust and quick viral detection system is essential for early, effective treatment.
In this case, ArcticZymes helped a biotech company to develop a highly sensitive and specific diagnostic kit by providing them valuable technical consultation from concept through commercialization of the PCR technology.
Making a better product
ArcticZymes’ partner was developing a diagnostic kit for the detection of enteroviruses. Several clinical studies have demonstrated the importance of the increased sensitivity and rapid turn-around time of reverse transcription PCR (RT-PCR) for EV detection. Building upon this knowledge, the idea was established to develop an efficient and user-friendly kit that offers high sensitivity and specificity in a single-tube format.
Since these kits were to be used in routine diagnostic hospital labs, the key concern was carry-over contamination from previous RT-PCRs. Traditional methods to prevent carry-over contamination would require costly investments such as separate lab spaces and personnel at each stage of the analysis.
With this in mind, the biotech company turned to ArcticZymes for help, relying on their expertise in the field of PCR technology, and years of experience in developing and manufacturing ultra-pure recombinant enzymes.
Together with ArcticZymes’ scientists, it was identified that the best method to reduce contamination risk would be a built-in control for carry-over contamination. The only known method for contamination control in PCR is the use of dUTP (uracil-containing nucleotide; U) in all PCR reactions in the laboratory followed by uracil-N-glycosylase (UNG), a selective degrader of uracil-containing DNA. The UNG enzyme is able to remove all copied DNA from previous reactions that may contaminate the sample before each new analysis.
However, most UNG enzymes are difficult to inactivate, requiring high temperatures that might be harmful to RNA. In addition, when compared to PCR, RT-PCR requires an additional step that converts the viral RNA to DNA, yet UNG treatment needs to be performed before RNA is converted to DNA (this step is called reverse transcription, or RT).
Crucially, RT usually occurs at 50°C, and at this temperature, traditional UNG enzymes are still active. Thus the UNG enzymes will destroy any DNA that is synthesized. Using traditional UNG products is considered impossible in RT-PCR applications. The solution recommended by ArcticZymes’ scientists was to use one of their unique enzymes, Cod UNG.
The perfect enzyme for the job
Cod UNG is active at low temperatures but is very sensitive to moderate heat. Unlike other UNGs that can be reactivated, Cod UNG is irreversibly inactivated. Thus Cod UNG-based decontamination can be carried out within the same RT-PCR tube. After obtaining and testing several batches of Cod UNG from ArcticZymes, our partner was able to incorporate Cod UNG into their viral detection kit, while maintaining the high sensitivity demanded and total assurance of eliminating the risk of any carry-over contamination. In additional, our partner found no differences in performance between batches, validating the enzyme’s reliability for diagnostic use.
ArcticZymes has built a very reliable reputation among its commercial customers for providing not only unique high quality enzymes, but also dedicated consultative support and a great customer service. On countless occasions, the assistance offered by ArcticZymes has successfully enabled it’s customers to market faster as well as saving precious time and resources.