Mycobacterium Tuberculosis - Diagenode Diagnostics

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PAGE 6 DIAGENODE MYCOBACTERIUM TUBERCULOSIS REAL-TIME PCR USER MANUAL CAUTION Diagenode commercializes a qualitative extraction and/or inhibition control. It is the responsible of the user to optimize the extraction procedure according to the result obtained for this extraction and/or inhibition control in order to get an appropriate result. • Virus control extraction must be manipulated using a safety cabinet for a manual extraction procedure or in a specific area for automated extraction procedures; as suggested by the manufacturer. Contamination • Perform nucleic acid release, isolation and real time amplification in separate laboratory areas. • Keep all tubes and vials closed when not in use. • The PCR laboratory (safety cabinet, DNA/RNA extraction platform, bench coat, etc.) must be cleaned after each PCR experiment with appropriate solutions. • The contaminated kits must be thrown in a specific waste bin for biological substances. • The kit should not be used by somebody displaying symptoms of the disease being detected by this kit. • The positive and negative PCR control must be performed with each reaction of PCR. • Gloves must be worn. • Aerosol resistant tips must be used for all PCR mixtures. • Tubes must be harvested before opening. • Do not pipette any of the materials by mouth. Do not smoke, eat or drink in areas which specimens or kit reagents are handled. • The experiment should not be carried out during pregnancy due to risks to the newborn baby. CAUTION Storage of the kit at room temperature can lead to the degradation of the primers / probes / positive control. This will lead to a decreased sensitivity and we would strongly advise a new kit. Material Required, Not Provided • Pipettes • Sterile pipette tips with filters • Powder-free gloves • Vortex mixer • Desktop centrifuge • RNase/Dnase-free microcentrifuge tubes (1,5 and/or 2 ml) • RNase/DNase free water • DNA isolation kit (see point II. Sample Extraction/DNA Isolation) • Consumables and accessories (reaction tubes, microcentrifuge, reaction plates, adhesive seal) for an ABI / Roche / Biorad / Qiagen / Stratagene or Cepheid system Europe Diagenode sa / CHU - Tour GIGA - B34 - 3 e étage // Avenue de l’Hôpital, 1 // 4000 Liège (Sart Tilman) // Belgium // Phone: (+32) 4 364 20 50 // Mail: info@diagenode.com
PAGE 7 • Master mix (see point III. PCR Reaction Set-up) • Real time PCR instrument: ABI / Roche / Biorad / Qiagen / Stratagene or Cepheid. Human sample used / Claims of the kit Mycobacterium tuberculosis Real-Time PCR kit is dedicated at the detection of the Mycobacterium tuberculosis in bronchoalveolar lavage, sputum samples. The kit is used on samples from potentially infected patients by Mycobacterium tuberculosis. Human samples’ Collection, Storage and Transport I. Samples’ Collection Human respiratory samples such as bronchoalveolar lavage, sputum should be collected. II. Samples’ Storage Human sample must be stored at 4°C for up to 24 hours or frozen at -20°C or -80°C for longer periods of time. Human samples should be stored in DNase/RNAse free polypropylene tubes. The sensitivity of the PCR could be reduced if you keep freezing and thawing human samples repeatedly. Avoid freeze / thaw cycles, which will affect the sensitivity of the assay. III. Samples’ transport Human respiratory samples must be transported in adapted containers. The samples must be transported following the local and national regulations for the transport of pathogen material. Human samples should be transported at -20°C. www.diagenodediagnostics.com
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Mycobacterium Tuberculosis - Diagenode Diagnostics

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