Multichannel: +7 (495) 984-69-93 Price list in PDF news About company Products Services Technical service Information support Vacancies Home > Products > Real Time PCR Reagent Kits FOR THE DIAGNOSIS OF TUBERCULOSIS Kits of reagents "Amplitub" for rapid diagnosis of tuberculosis, including multidrug-resistant and extensively drug-resistant (MDR-TB, XDR-TB). In 2004, on the initiative of Doctor of Medical Sciences, Professor Mikhail Aleksandrovich Vladimirsky, Head of the Molecular Diagnostics of Tuberculosis Sector of the Research Institute of Phthisiopulmonology of the I.M. IM Sechenov, we have begun development in the field of PCR diagnostics of tuberculosis. "Amplitub" is a technology for rapid molecular genetic diagnosis of tuberculosis for detecting mycobacteria of the tuberculosis complex, determining the infectious hazard of a patient, determining antibiotic resistance to drugs of I and II series, identification and genotyping of mycobacteria. The Amplitub technology consists of 3 main stages: sample preparation, DNA extraction and real-time polymerase chain reaction (PCR). The original technological developments of our company are used at each of these stages and together provide the highest sensitivity, specificity and information content of the analysis, the speed of obtaining the results and their high reliability. Stage I Sample preparation: during incubation with "Inactivating reagent A", mycobacteria are inactivated, dilution and homogenization of respiratory samples. Stage II DNA extraction: the use of original reagents and magnetic particles provides the maximum amount of pure DNA and allows you to automate the process of DNA extraction. III stage of real-time PCR: simultaneous detection of several different regions of the genome of the complex of mycobacterium tuberculosis (MBT) increases the information content and reliability of the analysis. Advantages of AMPLITUB technology Speed Amplitub technology reduces the time required to detect M. tuberculosis complex and determine drug resistance from 1-3 months to 1 day compared to traditional culture methods. High reliability of results High reliability of the results is ensured both at the stage of MBT detection and at the stage of determining antibiotic resistance. The use of Amplitub technology allows detecting MBT with high sensitivity (no more than 100 CFU / ml of sputum) and 100% specificity. Accurate identification of mutations in the most significant codons of the main genes associated with drug resistance to rifampicin and isoniazid makes it possible to determine 90% of antibiotic-resistant samples with a specificity close to 100%. Safety Clinical samples are inactivated with the "Inactivating reagent A" already at the stage of collection and transportation of biological material. Ability to work with clinical samples An important advantage of the "Amplitub" technology is the ability to study not only mycobacterial cultures, but also directly clinical material (sputum, bronchial lavage water, cerebrospinal fluid, exudate, punctate, biopsy, urine, genital tract discharge). Automation Robotic station TECAN and a set of reagents "M-Sorb-Tub-Avtomat" allow to fully automate the most time-consuming stage of DNA extraction. The Amplitub technology provides automatic calculation of the results of PCR analysis indicating the amount of the identified pathogen in the sample and specific mutations that determine antibiotic resistance. Stage I - Sample preparation inactivation “Inactivating reagent A” isspecially adapted for sputum processing. Possesses exceptional mucolizing properties, due to which it can be used instead of NALC-NaOH solution for sputum treatment before molecular genetic studies. Clinical and technical trials of Inactivating Reagent A were carried out at the Central Research Institute of Tuberculosis (Moscow). 118 sputum samples from severe bacillary patients with acute tuberculosis were divided into two parts, one part was treated with "Inactivating reagent A", the other - with NALC-NaOH. All samples were tested on BACTEC MGIT 960 and by real-time PCR (RT-PCR). Mycobacterial growth was observed only in the control group treated with NALC-NaOH. No growth of mycobacteria was detected in the samples treated with Inactivating Reagent A. The results of PCR analysis of samples from both groups were positive. Stage II - DNA isolation Synthol has developed a number of reagent kits for the isolation of mycobacterial DNA. "M-Sorb-Tub"- a set of reagents for the isolation of mycobacterial DNA from clinical samples and cell cultures (on magnetic particles). Adapted for the isolation of mycobacterial DNA from sputum, bronchial lavage water, cerebrospinal fluid, exudate, punctate, biopsy, urine, genital tract discharge, cell cultures. The M-Sorb-Tube kit provides cell lysis, DNA sorption on magnetic particles, DNA precipitation by centrifugation, DNA washing and elution. The DNA extraction method proposed by Syntol combines the advantages of the sorption method and the alcohol precipitation method. The use of magnetic particles coated with silica as a sorbent is more convenient in comparison with the use of other sorbents, it allows to standardize and automate the DNA extraction procedure as much as possible. An internal positive control (MIC) is added to each test sample, and the presence / absence of PCR inhibitors and the efficiency of DNA extraction are judged by the MIC amplification reaction. "M-Sorb-Tub-Avtomat" is a set of reagents for the isolation of mycobacterial DNA from clinical samples and cell cultures at automated TECAN stations. The kit is a modification of the "M-Sorb-Tube" reagent kit, designed specifically for use at automated stations of the TECAN company, which provides a number of the following advantages. Reducing the risk of contamination during DNA extraction to virtually zero. Significant reduction in the complexity of the DNA extraction process. Standardized DNA extraction from clinical specimens. Economic benefit: low consumption of plastic and reagents. Extra-Tube is a set of reagents for accelerated sample preparation and isolation of mycobacterial DNA from sputum samples. Combining the process of sample preparation and DNA extraction from sputum samples makes it possible to carry out these two stages of work in 1 hour. Minimal number of manipulations during sample preparation and DNA extraction. The set consists of 3 working solutions: "Inactivating reagent A", lysis and elution solutions. The use of "Inactivating reagent A" as part of the "Extra-Tube" kit allows increasing the DNA yield by 5-10 times. Stage III - real-time PCR Detection and quantification of the MBT complex TheAMPLITUB-RVreagentkitis: high reliability of the analysis due to the simultaneous detection of two specific genes of M. tuberculosis complex: IS6110 and regX3; assessing the suitability of a DNA sample for determining antibiotic resistance; high sensitivity (no more than 100 CFU / ml of sputum or 1-10 copies of DNA fragments in the reaction) and 100% specificity, confirmed on the QCMD panel (Quality Control for Molecular Diagnostics) Mtb EQA (UK). Determination of MBT antibiotic resistance to drugs of I and II series Reagent kits AMPLITUB-MDR-RV, AMPLITUB-XDR-RV are: диагностика форм туберкулеза с лекарственной устойчивостью, в том числе множественной лекарственной устойчивостью (МЛУ) и широкой лекарственной устойчивостью (ШЛУ); быстрая детекция мутаций M.tuberculosis, ассоциирующихся с устойчивостью к антибиотикам первого и второго ряда (рифампицину, изониазиду, этамбутолу, фторхинолонам); достоверность результатов, ежегодно подтверждаемая при проведении Федеральной службой внешней оценки качества (ФСВОК) Росздравнадзора испытаний шифрованных тест-культур и образцов. Differentiation within the M. tuberculosis complex AMPLITUB-DIFFERENTIATION is a set of reagents for the differentiation of mycobacterial species included in the M. tuberculosis complex (M. tuberculosis, M. bovis, M. bovis BCG and other species). Genotyping of mycobacteria AMPLITUB- Beijing - a set of reagents for determining the genotype Beijing of mycobacterium tuberculosis. The AMPLITUB-MDR-RV and AMPLITUB-XDR-RV kits are based on the use of an original multicompetitive allele-specific real-time PCR technique that allows detecting mutations in the genes of mycobacterium tuberculosis responsible for resistance to specific antibiotics. Принцип мультиконкурентной аллель-специфичной методики ПЦР в реальном времени. В формате одной пробирки используются 5’-флуоресцентно-меченые аллель-специфичные праймеры с общим комплементарным им 3’-меченым олигонуклеотидом-гасителем и контрольный флуоресцентный зонд, комплементарный участку ДНК без мутаций. При отсутствии мутаций в ДНК нарастание флуоресценции в ходе ПЦР-РВ наблюдается только по флуорофору зонда. При наличии мутаций в ДНК нарастание флуоресценции в ходе ПЦР-РВ наблюдается как по флуорофору зонда, так и по флуорофору 5’-флуоресцентно-меченого аллель-специфичного праймера. Данный метод позволяет определить не только точку мутации, но и процент устойчивого мутантного штамма МБТ на фоне дикого. Use of automated stations Freedom EVO (TECAN, Switzerland) for molecular genetic diagnosis of tuberculosis In 2011, at the initiative of the staff of the laboratory of molecular genetic research methods of the Central Research Institute of Research and Development of the Russian Academy of Medical Sciences, a group of developers from the Central Research Institute of Research under the leadership of Professor Larisa Nikolaevna Chernousova and "Synthol" for the first time in Russia introduced into laboratory practice a high-performance automated system for the isolation of mycobacterial DNA on the Freedom EVO platform (TECAN) using Amplitub reagent kits. This made it possible to significantly reduce the complexity of PCR diagnostics of tuberculosis and to increase the flow of analyzes. Time of MBT analysis when using automated stations Freedom EVO ("TECAN", Switzerland) and reagent kits "Amplitub" ("SYNTHOL", Russia) Labor costs - 1 hour Duration of DNA extraction - no more than 2 hours Preparation for amplification and RT-PCR - 1.5 hours TOTAL: the analysis results will be ready in 3.5 hours The operator will spend 1 hour of his working time. The structure of the automated station Freedom EVO ("TECAN", Switzerland) is shown below: Choosing Amplitub (Synthol) technology on the Freedom EVO (TECAN) platform, you get: Detection of the causative agent of tuberculosis - 48 samples in 3 hours Determination of drug resistance to drugs of I and II line in 1 day Reduction of labor costs by 6 times Increased laboratory productivity Work safety thanks to "Inactivating reagent A" Standardized DNA extraction Reducing the risk of laboratory contamination Low cost of 1 analysis Appreciation of phthisiatricians for fast and accurate diagnostics! Documents Download the brochure "Molecular Genetic Diagnosis of Tuberculosis by Real Time PCR" Download the brochure "Quickly Detect - Successfully Cure!" Back to section Products OLIGONUCLEOTIDES Modified oligonucleotides Real Time PCR Probes PCR mixtures for research laboratories Real Time PCR Reagent Kits Reagent kits for DNA and RNA isolation Sequencing reagents Reagents for genetic analysis Other reagents for molecular biology Equipment and consumables for PCR and genetic analysis Contacts 127550, Moscow, Timiryazevskaya 42, SINTOL Company Tel .: +7 (495) 984-69-93 multichannel +7 (499) 977-74-55 +7 (495) 506-79-97 Fax: +7 (495) 984-69-93 +7 (499) 977-74-55 E-mail: syntol@syntol.ru Ask a Question Website creation - © 2016 SYNTOL. All Rights Reserved news About company Products Services Technical service Information support Vacancies +7 (495) 984-69-93 E-mail: syntol@syntol.ru