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8 Laboratory Materials Contamination of continuous cell lines by mycoplasmata is a serious problem. As a chronic infection, it impairs the functions of cell cultures in many ways. They interfere with the metabolism of the cells altering their growth, their immunological and biochemical characteristics as well as their viability. This may have significant implications in terms of cost and time for the laboratories. An infection of a cell culture with mycoplasmata cannot be detected visually or under the microscope. It is therefore vital to carry out regular tests on the cell culture. The RIDASCREEN ® ‚ Mycoplasma Immunofluorescence Test contains highly specific monoclonal antibodies against a wide range of mycoplasmata, including the species Acholeplasma laidlawii, Mycoplasma hyorhinis, M. arginini, M. orale, M. fermentans and M. salivarium, which are known to be present in 96 % of all cell culture infections. The RIDASCREEN ® ‚ Mycoplasma IFA is carried out rapidly and is highly sensitive for the detection of mycoplasmata due to the application of a second FITC-labelled antibody. All reactants are ready-made and allow for two test methods: • One-Step Marking (Mark-Wash-Observe) For rapid screening of suspected positive cultures. • Two-Step Marking (Mark-Wash-Mark-Wash-Observe) For cultures with doubtful or slightly positive initial results. After the first step, a FITC-labelled anti-MAK antibody is introduced; incubate for a further 20 minutes at room temperature. Results Yellow-green fluorescence on the rim or between the redcoloured cells. Comparative studies of Different Methods for the Detection of Mycoplasmata in Cell Cultures 8.1 Labormaterialien Mycoplasma Detection/Mycoplasmen-Nachweis Description/Beschreibung Mycoplasmen stellen als Kontaminanten in kontinuierlichen Zelllinien ein großes Problem dar. Als chronische Infektion beeinträchtigen sie auf vielfältige Weise die Funktion von Zellkulturen. Sie nehmen Einfluss auf den Metabolismus der Zellen und verändern so deren Wachstum, ihre immunologischen und biochemischen Eigenschaften sowie deren Lebensfähigkeit. Dies bedeutet Geld- und Zeitverlust für Zellkultur-Laboratorien. Die Infektion einer Zellkultur mit Mycoplasmen ist visuell oder lichtmikroskopisch nicht zu erkennen. Aus diesem Grunde ist eine regelmäßige Überprüfung einer Zellkultur wichtig. Der RIDASCREEN Mycoplasmen Immunfluoreszenztest enthält monoklonale Antikörper mit hoher Spezifität gegen ein breites Spektrum von Mycoplasmen, einschließlich der Spezies Acholeplasma laidlawii, Mycoplasma hyorhinis, M. arginini, M. orale, M. fermentans und M. salivarium, welche zu mehr als 96 % an Zellkulturinfektionen beteiligt sind. Der RIDASCREEN Mycoplasma IFA ist schnell durchzuführen und durch die Verwendung eines FITC-markierten zweiten Antikörpers hoch sensitiv für den Nachweis von Mycoplasmen. Alle Reagenzien sind gebrauchsfertig und erlauben zwei Arten der Durchführung: • Ein-Schritt-Färbung (Färben-Waschen-Betrachten) Zum schnellen Screening wahrscheinlich positiver Kulturen • Zwei-Schritt Färbung (Färben-Wachen-Färben-Waschen-Betrachten) Für fragliche oder schwach positive Ergebnisse. Nach dem ersten Schritt wird ein FITC-markierter Anti- MAK Antikörper zugegeben und weitere 20 min bei Raumtemperatur inkubiert. Ergebnisse Gelbgrüne Fluoreszenz am Rand oder zwischen den rot gegengefärbten Zellen. Vergleichende Untersuchungen verschiedener Methoden zum Nachweis von Mycoplasmen in Zellkulturen Test method/Testsystem Sensitivity/Sensivität Specifity/Spezifität DNA-RNA-Hybridisation/DNA-RNA-Hybridisierung 100 % 100 % PCR (in-house) 100 % 100 % ELISA 65,5 % 100 % DNA-Marking (direct)/DNA Färbung (direkt) 93,1 % 92,3 % RIDASCREEN ® Mycoplasma IFA 100 % 92,3 % Reference: from culture 42 cell lines (29 pos./13 neg.) of different origin were tested. Referenz: kulturelle Anzucht Es wurden 42 Zelllinien (29 pos./13 neg.) unterschiedlicher Herkunft untersucht. BIOTECH GmbH
Laboratory Materials Mycoplasma Detection • Ready-made reagents • Contains cell stainer • Dropping bottle • Controls included • Rapid processing • Rapid • Safe • Sensitive • Reliable RIDASCREEN ® ‚ Mycoplasma IFA* • A new immunofluorescence test kit. • A new method of mycoplasma - detection in cell cultures, using monoclonal antibodies. * registered trademark BIOTECH GmbH 8.2 Labormaterialien Mycoplasma Detection/Mycoplasmen-Nachweis Advantages/Vorteile Mycoplasmen-Nachweis • Gebrauchsfertige Reagenzien • Gegenfärbung enthalten • Tropffläschchen • Kontrollen eingeschlossen • Schnelle Durchführung • Schnell • Sicher • Sensitiv • Zuverlässig RIDASCREEN ® ‚ Mycoplasmen IFA* • Ein neuer Immunfluoreszenz-Test. • Eine neue Methode zum Mycoplasmen-Nachweis in Zellkulturen unter Verwendung monoklonaler Antikörper. * eingetragenes Warenzeichen Mycoplasma-detection for 20 tests 1 x R-4203 Mycoplasmen-Nachweis für 20 Bestimmungen Mycoplasma-detection for 50 tests 1 x R-4202 Mycoplasmen-Nachweis für 50 Bestimmungen 8
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Sera Other Animal Sera/Andere Tiers
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Cell Culture Media Zellkulturmedien
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Media MEM Eagle with EBSS 500 ml P0
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Reagents Reagenzien Media Supplemen
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Services Protein Colorimetric test
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Biologicals Biologika Introduction
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