Hzymes SFAC-LAMP Molecular Diagnostic Program
COVID-19 has put molecular diagnostic products around the world through an unprecedented test. In the event of human outbreaks and animal disease inspections, the demand for molecular diagnostic POCT products will continue to expand. The demand for simple, fast, accurate, and compatible molecular diagnostic POCT products has been highlighted under the normalized epidemic prevention mechanism. According to the situation of normalization of global epidemic prevention, Hzymes Biotech launched the overall solution of SFAC-LAMP molecular diagnostic raw materials and applications, providing simple, fast, accurate and compatible for application scenarios such as grass-roots medical institutions, airports, customs inspection and quarantine, animal epidemic prevention real-time detection, etc. technical solutions.
The full name of LAMP is Loop-mediated Isothermal Amplification, that is, loop-mediated isothermal amplification reaction, which belongs to a kind of constant temperature amplification technology. Its biggest technical feature is that it does not require temperature rise and fall, and its dependence on instruments is very low. At the same time, by adding visualization dyes, the results can be directly observed with naked eyes. Hzyme provides a complete set of solutions for LAMP technology, from primer design optimization to product detection, a one-stop solution for the development process of molecular diagnostic POCT reagents. At 65°C, it only takes 15 minutes to get a positive result for 100 copies, and 20 minutes for 10 copies. The detection range is from 20 to 106 copies. It can still be detected after 8 days of thermal acceleration at 37°C.
In order to solve the most difficult aspect of POCT reagent for molecular diagnostics and guarantee the accuracy of testing results, Hzymes Biotech has improved the detection sensitivity (<10 copies) and stability (no significant change in reagent performance after 37℃thermal acceleration and 7-day continuous testing or 50℃ thermal acceleration and 48-hour continuous testing) of all aspects (e.g. primer optimization, raw material preparation) of LAMP technology platform.
Especially at the stage of researching and developing raw materials, we have improved the enzymatic activity and stability of core raw materials including BstL DNA Polymerase, RTL Transcriptase, and RNase Inhibitor via molecular modification and structural improvement. Therefore, Hzymes has not only ensured the accuracy of detection results, but also enhanced system stability.
Bst DNA polymerase features:
Bst 2.0 DNA polymerase features:
Compared with similar products, Bst DNA polymerase V2 has a balanced false positive and peak time, and has good stability in thermal acceleration. It greatly saves time and cost while the results are reliable. Hzyme also provides a high-concentration version of Bst DNA polymerase V2 to meet customers’ freeze-drying needs. RTL reverse transcriptase RTL reverse transcriptase can be used for first-strand cDNA synthesis. The reaction buffer is optimized for RT-LAMP. It is highly compatible with Bst DNA polymerase V2. It is thermally accelerated at 37°C for 7 days and has good stability. It has excellent performance in the RT-LAMP reagent system, which is equivalent or even better than imported competing products. At the same time, a high-concentration version of RTL is also provided to meet the freeze-drying needs of customers.
RNase inhibitor
The activity of Hnzymes RNase inhibitor was not affected by freezing and thawing 10 times, and the activity was still stable after being treated at 55°C for 5 minutes. Compared with world-class enzyme products, the stability and inhibition performance are better.
RT-LAMP nucleic acid amplification reagent (red-yellow color method)
The accelerated stability at 37°C is good, and the reaction condition is 65°C for 30min. As shown in the upper right figure, heating at 37°C for 4 days has no effect on the performance of the reagent, and heating for 8 days still does not affect the detection of positive samples. It can detect samples with a large concentration range, and the unit of sample concentration is copy/reaction, as shown in the lower right figure, it can be detected from 20-106 copies/reaction.
RT-LAMP
The RT-LAMP COVID-19 Detection Kit has high sensitivity. According to customized requirements, it can detect samples with a minimum concentration of 1000 copies/mL (50 μL system) or 500 copies/mL (100 μL system). Compared with competing product N, with the same template concentration of 50copies/T, the detection time of Hzymes RT-LAMP is shorter.
Through the research and development team of Hzyme, we can now provide FASC-LAMP solutions. The products include RT-LAMP Visual Detection Kit (red and yellow) and RT-LAMP Visual Detection Kit (fluorescent method). Both products are compatible with RNA and DNA testing solves multi-product pipeline problems at one time. Good compatibility can ensure the simplification and versatility of products in multiple application scenarios, thus providing more possibilities for the practical application of LAMP technology.
