Solutions for Veterinary Testing

Transcription

1 Solutions for Veterinary Testing Sample & Assay Technologies

2 Your partner for veterinary testing At QIAGEN, we strive to provide veterinary testing professionals with advanced sample and assay technologies. The demand for nucleic acid based testing for veterinary pathogens is increasing due to the speed and high sensitivity of these methods. We are committed to drive and support this development. Technology leadership As the market and technology leader in molecular sample and assay technologies, we provide integrated, standardized systems required for reliable and sensitive veterinary testing. QIAGEN s advanced sample and assay technologies are used by veterinary laboratories around the world and are part of standard protocols recommended by numerous government agencies. Setting standards in reference laboratories National reference laboratories rely on QIAGEN solutions for the detection and analysis of many economically important animal diseases including Bluetongue virus (BTV), classical swine fever (CSF), bovine viral diarrhea (BVDV), and foot and mouth disease. QIAGEN cador real-time PCR kits for BVDV testing are approved by the Friedrich-Loeffler-Institut (FLI) in Germany and the Swiss government and are licensed for use in the Swiss BVDV eradication program. QIAGEN continues to meet regulatory needs around the world, providing you with validated solutions in compliance with national and international regulatory requirements. Complete solutions from sample to assay Combining sample preparation and assay technologies, QIAGEN provides integrated, standardized systems for reliable veterinary testing. Sample collection and disruption technologies enable access to the most challenging animal samples. Trusted QIAGEN purification methods ensure maximum yield of highly pure nucleic acids from host and pathogen genomes. Flexible automated solutions increase standardization and streamline laboratory workflows. Together with our assay technologies, QIAGEN provides a complete, customizable solution for molecular veterinary laboratories. QIAGEN Quality. In an independent survey (Customer Loyalty & Satisfaction, BioInformatics [2006]), more than 1000 scientists highlighted QIAGEN for an excellent quality of products and services from highly consistent product performance to excellent customer service and convenient ordering systems all over the world. Partnering with the experts QIAGEN understands the importance of reliability in modern veterinary testing and has partnered with leading veterinary laboratories, including the Veterinary Laboratories Agency (VLA) and the Institute for Animal Health (IAH) in the United Kingdom and the Institute of Veterinary Virology (IVV) in Switzerland, to develop and validate technologies for veterinary testing. The partnership between the VLA and QIAGEN combines VLA s in-depth expertise in animal diseases and veterinary diagnostics with QIAGEN s strong technological capabilities and access to veterinary testing labs around the globe. The VLA is pleased to be able to share through the QIAGEN partnership with laboratories across the world ready-to-use, validated PCR assays that VLA has developed to support the UK livestock industry and Government animal health programs, Roger Hancock, Veterinary Director of VLA 2 Sample & Assay Technologies Solutions for Veterinary Testing 03/2009

3 Efficient access to analytes is a prerequisite when dealing with animal tissues, blood, bacteria, and insects. QIAGEN provides convenient sample disruption solutions for successful animal molecular testing. Our sample archiving solutions allow easy access to the DNA even years later. QIAGEN sample-collection technologies provide: Direct sample collection on QIAcard FTA Spots Stability at room temperature for years Convenient DNA purification protocols The QIAcard FTA procedure. Simply apply the sample to the QIAcard FTA Spot. DNA is stable for years at room temperature. QIAGEN sample-disruption technologies provide: Flexible disruption solutions for low to high throughput Highly standardized methods for reproducible results Proven results with animal, insect, bacteria, and yeast samples The TissueLyser II. For simultaneous disruption and homogenization of up to 2 x 96 samples. Case studies: Sample collection Bluetongue viral RNA detection: Rapid and efficient disruption of midges to access Bluetongue viral RNA is successfully achieved using the TissueLyser II (1). CSF RNA detection: Swine tissue samples including lung, heart, spleen, lymph node, kidney, tonsil, muscle are disrupted and homogenized using the TissueLyser II. Taylorella equigenitalis bacterial DNA detection: No disruption required. BVDV RNA detection: The TissueLyser II can be used to disrupt earnotch samples. 1. Veronesi, E., et al. (2008). J. Med. Entomol. 45, 129. Proceed with purification 3 Solutions for Veterinary Testing 03/2009

4 The quality of the purified nucleic acid greatly influences the reliability and sensitivity of the subsequent PCR assay. QIAGEN s nucleic acid purification portfolio provides optimized solutions for a variety of animal sample types. QIAGEN products are used by reference laboratories throughout the world. Multiple citations demonstrate how much veterinary molecular testing professionals rely on QIAGEN. Automation platforms from QIAGEN provide: Automated solutions for every throughput need Instruments developed in conjunction with purification technologies Flexible, standardized systems The QIAcube. Easy automation of QIAGEN spin-column kits. The BioSprint 96. Semiautomated nucleic acid purification for up to 96 samples. IQ/OQ and validation support services Comparable performance across platforms for rapid adjustments in throughput The QIAsymphony SP. Flexible, automated, medium- to high-throughput sample preparation for 1 96 samples. The BioRobot Universal System. Automated high-throughput nucleic acid purification in 96-well format. Nucleic acid purification kits from QIAGEN provide: Purified DNA or RNA ready for reliable and sensitive PCR Wide range of kits optimized for various sample types Dedicated kits for pathogen nucleic acid isolation Gold standard QIAamp, DNeasy, and RNeasy spin-column kits Innovative magnetic-particle technology for high throughput BioSprint 96 One-For-All Vet Kit for high-throughput nucleic acid purification from a wide range of veterinary sample types QIAamp, DNeasy, and RNeasy Kits. Spin-column technology trusted for years by veterinary molecular testing professionals. Case studies: Purification Bluetongue viral RNA detection: Medium-throughput Bluetongue viral RNA purification from cattle blood using the QIAcube, or high-throughput purification using the BioRobot Universal System (Figure 1, page 6). CSF RNA detection: Cleared lysates are processed using the RNeasy Mini Kit manually or on the QIAcube. Taylorella equigenitalis bacterial DNA detection: DNA is isolated from bacterial cultures using the QIAamp DNA Mini Kit on the QIAcube. BVDV RNA detection: Blood and tissue samples are further processed on the QIAsymphony SP. Proceed with analysis 4 Solutions for Veterinary Testing 03/2009

5 QIAGEN provides a range of solutions for sensitive and reliable PCR analysis. We have the assays, enzymes, and instruments you need for pathogen detection and genotyping. QIAGEN is also one of the few companies with a worldwide PCR license for veterinary testing, without additional licensing costs for the user. Ready-to-use cador real-time PCR kits are developed for sensitive detection of specific animal pathogens. QIAgility automated PCR setup in all formats: Convenient, easy-to-use software Standardized results and increased productivity The QIAgility eliminates manual pipetting steps. The cador BVDV Type 1/2 RT-PCR Kit. The kit is designed to differentiate BVDV 1, BVDV 2, and border disease virus (BDV), a related pestivirus, in samples from cattle. cador real-time PCR kits provide: Assays developed in cooperation with leading animal health authorities High reliability with internal and external controls* QIAGEN real-time and end-point PCR and RT-PCR kits provide: A comprehensive portfolio, from fast cycling to multiplex PCR QuantiTect Virus Kits, for singleplex or multiplex virus detection Type-it Kits, optimized for genotyping applications QIAxcel System automated analysis of DNA fragments and RNA: The Rotor-Gene Q, QIAGEN's real-time PCR cycler, for ultrafast, high-precision analyses. Accurate analyses with high resolution Streamlined workflow minimizes analysis time Rotor-Gene Q real-time PCR cycler in rotary format: Outstanding thermal and optical performance * The cador BVDV Type 1/2 RT-PCR Kit is designed to differentiate types of BVDV in samples that have already tested positive. Therefore the kit does not include an internal control. Unmatched optical range from UV to infrared Case studies: Analysis Bluetongue viral RNA detection: The cador BTV RT-PCR Kit (co-developed with the IAH) uses a novel molecular assay to detect 24 known strains. CSF RNA detection: Quantitative real-time RT-PCR (1) using the QuantiTect Probe RT-PCR Kit (Figure 2). Taylorella equigenitalis bacterial DNA detection: Reliable detection of Taylorella equigenitalis bacterial DNA with the cador T. equigenitalis PCR Kit (Figure 3). BVDV RNA detection: Reliable detection with the cador BVDV RT-PCR Kit on the Rotor-Gene Q (Figure 4). 1. Hoffmann, B., et al. (2005). J. Virol. Methods. 130, 36. See next page for results Solutions for Veterinary Testing 03/2009 Sample & Assay Technologies 5

6 Reliable results from sample to data Veterinary laboratories around the world rely on a wide range of QIAGEN sample and assay technologies. The data below is representative of pathogen testing results from leading veterinary laboratories evaluating QIAGEN systems. Case studies: Results A β-actin: C T = N 9: C T = 25.7 N 4: C T = 33.9 β-actin N 9 N 4 B β-actin: C T = 21 N 19: C T = 26.6 N 291: C T = 32.3 β-actin N 291 Figure 1. Comparable performance for BTV detection from A medium throughput on the QIAcube to B high throughput on the BioRobot Universal System. No differences in sensitivity and specificity of the assay were observed. The implementation of different automated procedures increased the throughput capacity to approximately 470 samples/day and facilitated standardization and tracking of results. This method has been successfully transferred into routine workflows and provides the necessary flexibility for different throughput needs. The internal control is β-actin; N 9, N 4, N 19, and N 291 are BTV-specific targets. (Data kindly provided by Chantal Audeval, Magali Ackermann and Françoise Colocci, LDA 58, Nevers, France) N 19 C value T Lung Heart QIAcube Manual Spleen Lymph node - head Lymph node - abdominal Kidney Sample type Tonsil Muscle Δ Rn Dilution 1:10 1 1:10 2 1:10 3 1:10 4 1:10 5 1:10 6 1: Cycle number Figure 2. Efficient purification and sensitive detection of CSF viral RNA from different tissues. Tissues were disrupted and homogenized using the TissueLyser II. Viral RNA was purified using the RNeasy Mini Kit, either manually or automated on the QIAcube. Classical swine fever virus (CSFV) RNA was detected by quantitative, real-time RT-PCR using the QuantiTect Probe RT-PCR Kit. (Data kindly provided by C. Korthase and B. Hoffmann, Friedrich-Loeffler-Institute, Germany) Figure 3. Sensitive and reliable detection of T. equigenitalis DNA over a broad dynamic range. T. equigenitalis DNA was purified from bacterial culture dilutions using the QIAamp DNA Mini Kit on the QIAcube. T. equigenitalis DNA was detected using the cador T. equigenitalis PCR Kit on the Applied Biosystems 7500 Real-Time PCR System. (Data kindly provided by J. Errington and P. Wakeley, Veterinary Laboratories Agency, UK) :4 dilution Undiluted Δ Rn Threshold Cycle number Figure 4. Isolation of BVD virus from a positive blood sample. Viral RNA was isolated from a 200 µl blood sample from infected cattle, either undiluted or a 1:4 PBS dilution using the QIAsymphony SP. A 10 µl aliquot of each eluate from four purification replicates was used for real-time PCR analyses with the cador BVDV RT-PCR Kit on a Rotor-Gene real-time cycler. Solutions for Veterinary Testing 03/2009 Sample & Assay Technologies 6

7 Expert service and support Global service and support QIAGEN s veterinary testing experts provide exceptional service and technical support. Specialists in technical services and sales are extensively trained in veterinary testing applications. QIAGEN also provides guidelines and on-site support to fast-track the validation of new methods. Experienced instrument service QIAGEN Instrument Service provides comprehensive support services. Application Services and Training Programs give the freedom and flexibility to adapt systems to the specific and changing testing needs found in veterinary laboratories. ISO 9001/ISO certification and an international team of highly qualified and experienced Support Specialists guarantee high-quality service. Application support Application laboratories in Germany, the USA, and Singapore provide a range of expert services, including on-site demos, validation support, and protocol adaptations. Training programs are available for automation of veterinary testing applications, tailor-made to meet customer needs. Expert PCR training Veterinary PCR training at QIAGEN provides both theoretical and practical training on major real-time PCR cyclers. Theoretical training includes an introduction to pathogen detection, key features of cador PCR Kits and QIAGEN PCR enzymes, programming of thermal cyclers, and general troubleshooting. Hands-on practical training in the laboratory includes sample preparation, pathogen detection and quantification plus detailed analysis of results. Visit our animal and veterinary testing resource pages at cador BVDV PCR Kits are approved by the Friedrich-Loeffler-Institute in Germany for veterinary diagnostic use. For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user manuals are available at or can be requested from QIAGEN Technical Services or your local distributor. Trademarks: QIAGEN, QIAamp, QIAcard, QIAcube QIAGEN Quality, QIAgility, QIAsymphony, BioRobot, BioSprint, cador, DNeasy, QuantiTect, RNeasy (QIAGEN Group); Applied Biosystems (Applera Corporation or its subsidiaries); FTA (Whatman Group); Rotor-Gene (Corbett Research) QIAGEN, all rights reserved. Solutions for Veterinary Testing 03/

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