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17 September 2018

Sequoia Genetics GC Alkor Bio received a patent for the invention "The method of specific identification of DNA sequences"

In the beginning of September, Sequoia Genetics Company (part of the Alkor Bio Group of Companies) received a patent for the invention "Method of Specific Identification of DNA Sequences". The patent No. 2665631 was issued on September 3, 2018 by the Federal Service for Intellectual Property in Russian Federation (Rospatent). Also the patent application was filed under the PCT system and was published with number WO 2018/139955 in August 2018.

The patent was received within the framework of the innovation project "Multiplex duplex-mediated analysis based on cleavable probes", which Sequoia Genetics develops with the support of the Skolkovo Foundation. Before, in 2016 Sequoia Genetics Company (GC Alkor Bio) received a grant from the Skolkovo Foundation in the amount of 22.5 million rubles for the development of this project. The project will be implemented in the field of molecular diagnostics based on polymerase chain reaction (PCR) technology. Multiplex duplex-mediated analysis based on cleavable probes is a platform for development diagnostic tests in a multiplex format. With this solution, Sequoia Genetics is going at least 3 times increase the multiplexing rate (in comparison with the standard technologies already being implemented in the field) for carrying out qualitative PCR-based analysis with real-time detection.

Senior Project Manager "Multiplex duplex-mediated analysis based on cleavable probes", author of the patent Vitaly Vedernikov:

"Automation and standardization, facilitation and acceleration of the analysis are the main directions of evolution in laboratory diagnostics. One of the ways to simplify and accelerate analysis is the development of multiplex technologies. The main advantages of multiplex analysis are:

 1. Reduced labor costs;

 2. Increase the speed of the analysis;

 3. High reproducibility of the analysis (1 operator, 1 method, 1 instrument, simultaneous analysis);

 4. Saving of reagents;

 5. Saving of biomaterial.

 Multiplex technologies have been used for a long time, including polymerase chain reaction (PCR). PCR with the real-time detection of the products (Real-Time PCR) due to the possibility of quantitative assessments, high specificity and sensitivity is one of the widely used technologies for tackling various genetic diagnostic problems, it can be performed in multiplex format too.

The widespread methods based on real-time PCR for simultaneous identification several targets in one tube do not provide a high degree of multiplexing since in one channel of the cycler is detected one target. Thus, the promising trend in the development of PCR technology is the development of approaches that increase the degree of multiplexing of the diagnostic test without complicating the procedure and lowering its sensitivity.

The aim of the project is the development and commercialization of the technology, representing a platform for creating diagnostic tests with a high degree of multiplexing (in one channel of the cycler is detected- 3 targets or more). The project is meant to create a comprehensive solution for a wide range of molecular genetic diagnosis problems. The platform consists of 1) set of artificial nucleotide sequences representing the basis of the technology and 2) software for interpreting the results of the analysis. The proposed solution is based on wide spread Real-Time PCR technology, i.e. the analysis does not require special equipment to be used, that can ease up market entrance. At the moment, a diagnostic test is being developed on the basis of the platform to identify 14 targets in one tube. The proposed platform is universal, accordingly, the project can be important in the molecular diagnostic (MDx) / in vitro diagnostic (IVD) segment by the developers of diagnostic kits. Tests developed on the basis of the platform will be demanded by diagnostic centers.

The innovative approach consists in an original detection scheme based on PCR, followed by melting analysis.

The gist of the approach is as follows: during the PCR, specific probes are split into duplexes. After the PCR, the reaction products are melted. By the presence of a duplex with a certain melting point, one can judge the presence of an appropriate target. The melting point of the corresponding duplex is specified by the developer for each target individually. The degree of multiplexing (the number of detectable targets in one tube) depends on the number of channels of the amplifier and on the number of duplexes that differ in melting point per channel. In this way, 2D target encoding is performed where the variable parameters are the melting point of the duplex and the amplifier channel. For example, for a 5-channel amplifier in one tube, 30 targets can be determined, when six targets are detected in one channel (see Fig.)».