Our Company. CerTest Biotec is a company focused on the development and manufacture of in vitro diagnostic products for human applications.
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2 Our Company CerTest Biotec is a company focused on the development and manufacture of in vitro diagnostic products for human applications. The company was founded in 2002 and its success is highly associated to three basic pillars, our human resources, innovation and customer orientation. These conditions allow us to provide our customers with products for clinical diagnosis with high standards of quality, more than 120 countries across the 5 continents. VIASURE Real Time PCR Detection Kits
3 Molecular Diagnostics Molecular detection techniques have revolutionized clinical diagnosis over the past several years. In particular, the Real Time Polymerase Chain Reaction has been turned into one of the most powerful in vitro diagnostic tools for the detection VS - ATV113 product strips low (L) / high (H) code or plate In this way, the Real Time PCR allows us to identify the causal VIASURE IC VIASURE was created with the purpose to provide Molecular Diagnostic users new Real Time PCR kits for the detection of pathogens of interest. VIASURE and compatibility with leading open system thermal cyclers and the latest technology for the stabilization of products. signal signal IC VIASURE Real Time PCR Detection Kits are designed for the diagnosis of infectious diseases caused by different pathogens in human samples. This new product range is based on 5 nuclease chemistry. This technology utilizes two quantity of target sequence present in the sample and could be measured on a wide range of Real Time PCR platforms. and Internal Control VIASURE Real Time PCR Detection Kit contains in each well all the necessary components for Real Time PCR assay in a stabilized format, which allows the shipment and storage at room temperature. General Catalogue
4 Work Flow Determine and separate the number of required reactions including samples and controls. One positive and negative control must be included in each run. Peel off protective aluminium seal from plates or strips. Step 1 Step 2 Step 3 Step 4 Add 15 l of rehydration Add 5 Load the strips into Interpretate results. buffer into each well. the thermocycler or Positive control, protocol. Components Reagent / Material Description Quantity 8-well strips 96-well plate and internal control in stabilized format and internal control in stabilized format 1 plate Rehydration Buffer Solution to reconstitute the stabilized product Positive Control 1 vial free Tear-off 8-cap strips Optical caps for sealing wells during thermal cycling Shell Frame Grid Shell Frame Grid 1 or 2 Components included for each VIASURE Real Time PCR Detection Kit. Kit References Reference VS - XXX106L VS - XXX106H VS - XXX112L VS - XXX112H VS - XXX113L VS - XXX113H Description Viasure (name of the product Viasure (name of the product Viasure (name of the product Viasure (name of the product Viasure (name of the product Viasure (name of the product Change XXX for the reference code of the selected kit. VIASURE Real Time PCR Detection Kits
5 and Advantages Ready-to-use Kits, containing all the necessary components for the testing. Transport and storage at room temperature. -for all our qpcr products-. Easy-to-use. It minimizes the number of manipulations reducing time and possible errors. early stages of the infection. Versatile. Possibility to analyze from 1 up to 96 samples per assay. Evaluated and validated according to ISO and CE marked. Compatibilities Manufacturer Model Qiagen Rotor-Gene Q* Cepheid SmartCycler * Manufacturer Model Qiagen Rotor-Gene Q* Cepheid SmartCycler * DTprime Real-time Detection Thermal Cycler DTprime Real-time Detection Thermal Cycler DTlite Real-time PCR System DTlite Real-time PCR System 7500 Fast Real-time PCR System 7300 Real-time PCR System 7500 Real-Time PCR System QuantStudio TM 7900 HT Real-Time PCR System QuantStudio TM QuantStudio TM ABI PRISM 7000 ABI PRISM 7700 QuantStudio TM 3 Real-Time PCR System QuantStudio TM QuantStudio TM 5 Real-Time PCR System QuantStudio TM StepOne Plus TM Real-Time PCR System QuantStudio TM StepOne TM Real-Time PCR System QuantStudio TM 3 Real-Time PCR System ViiA TM 7 Fast Real-Time PCR System QuantStudio TM 5 Real-Time PCR System CFX96 Touch TM Real-Time PCR Detection System Viia TM 7 Real-Time PCR System Mini Opticon TM Real-Time PCR Detection System CFX96 Touch TM Deep Well Real-Time PCR Detection System Roche LightCycler 480 Real-Time PCR System icycler iq TM Real-Time PCR Detection System Roche LightCycler 96 Real-Time PCR System icycler iq TM 5 Real-Time PCR Detection System Agilent Technologies MyiQ TM Real-Time PCR Detection System TM 96 (Fast block) MyiQ TM 2 Real-Time PCR Detection System Eppendorf Mastercycler TM ep realplex Technologies TM Real-Time PCR System compatibility, see instructions for use. * The product should be reconstituted following the appropiate procedure and Rotor-Gene or SmartCyclers tubes. Technologies TM Real-Time PCR System Analytik Jena Biometra TOptical Analytik Jena Biometra qtower 2.0 Abbott Abbott m2000 RealTime System TM General Catalogue
6 Real Time PCR Initiation Protocol VIASURE Real Time PCR Detection Kits
7 The objective of this protocol is to facilitate the training of laboratory technicians to perform molecular diagnostic test based on Real Time PCR technology or qpcr from clinical samples. The in combination with the signs and symptoms presented by the patient, will allow health personnel to stablish a clinical diagnosis. For the right development of the mentioned activities, it is necessary to adapt the facilities with adequate equipment and stablish different areas where every stage of the process is going to be Nucleic acids extraction area results interpretation area Sample processing. Reconstitute the required number of wells. Reconstitute lyophilized positive control. qpcr equipment. with a manual kit or automatic method Add negative control. Program the thermal cycler and start the (optional). protocol Add positive control. Close the wells with optional caps. Analysis of results with the software installed in the equipment. Recommendations necessary to keep standards of hygiene, order and security, in order to avoid possible contaminations that could seriously affect the right development of the test. Frequent use of disinfectants at the early stage of the process and after using the equipment. Commercial disinfectant solution. 70% ethanol. General Catalogue
8 1 Nucleic acids extraction area You can use your optimized manual or automatic system. Follow the manufacturer s instructions, or ask for 2 Pre-PCR area should be maintained free from any biological material to avoid possible contamination that could interfere with test results. disposable gloves, lab coat, mask, cap. Micropipette (2-20 μl) the wells and adding the negative control). Filter tips. Scissors ml test tube racks. area. wells (including samples and the two saved wells for the positive and negative controls that must always be included in each assay). If necessary, cut the required number of wells with scissors. Remove the protective aluminium seal. Reconstitute each one of the wells with 15 μl of rehydration buffer (blue vial). Add 5 μl of negative control (purple vial) in the reserved well for negative control. After this process, move the rehydrated wells VIASURE Real Time PCR Detection Kits
9 3 Biological risk is not potentially infectious for the laboratory staff, but it is recommended to keep the area clean and disinfected to avoid any contamination that could compromise the test result. area could be shared with the pre-pcr area (area 2) cleaning properly the cabinet between both processes and differentiating the use of the pipettes (1 st control, 2 nd disposable gloves, lab coat, mask, cap. Micropipette (2-20 μl) of nucleic acids and positive control). Filter tips. Scissors ml test tube racks. area. The VIASURE Pathogen Real Time PCR must be rehydrated before using in the assay. Since it contains a high number of template copies of the pathogen, it is recommended to open and manipulate before starting with the assay. After that, clean the area with Reconstitute the positive control (red vial) Divide in aliquots to minimize the freeze-thaw cycles. Store at -20ºC after re-suspension. sample in the rehydrated wells (with the positive and negative control). Add 5 μl of reconstituted positive control (red vial) in the reserved well for positive control. Divide the required optical caps number and close the wells. If necessary, cut the required optical caps number with a scissor. Be careful not to touch inside with the gloves. Real Time PCR equipment. General Catalogue
10 4 and results interpretation area This area is where the equipment and related components are located. It is recommended to place this area Real Time PCR equipment. Computer. of the thermocycler block to balance the equipment. Program the thermocycler and start protocol. area. DNA THERMAL CYCLING CONDITIONS RNA THERMAL CYCLING CONDITIONS Cycles Time Temperature 1 2 min 95 ºC 10 s 95 ºC s 60 ºC * * Data collection Cycles Time Temperature 1 15 min 45 ºC 1 2 min 95 ºC 10 s 95 ºC s 60 ºC * Internal Control Negative Control Positive Control Interpretation + + Positive pathogen Invalid Invalid * Detection of internal control is not necessary for highly positive samples. High copy number of target in the sample can lead to a weak or absent signal in the internal control. VIASURE Real Time PCR Detection Kits
11 CerTest Biotec, S.L , San Mateo de Gállego, Zaragoza (Spain) VIASURE online
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