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• W1996776500 abstract "<b><i>Background: </i></b>Contamination of cell culture and biological material by mollicute species is an important safety issue and requires testing. We have developed a singletube real-time polymerase chain reaction (PCR) assay for rapid detection of <i>Mollicutes</i> species stipulated by the European Pharmacopeia. <b><i>Methods: </i></b>Primers and TaqMan probes (FAM-labeled) were deduced from 16S rDNA sequence alignment of 18 mollicutes species. A synthetic internal control (IC) DNA and an IC-specific TaqMan probe (VIC-labeled) were included. The analytical sensitivity of the assay was determined on DNA dilutions from 12 mollicute strains. Specificity was proven by the use of DNA from other bacteria. <b><i>Results: </i></b>Analytical sensitivities of the PCR assay were in the range of 405-2,431 genomes/ml for 11 of the 12 tested mollicute DNA samples. The lowest sensitivity was found for <i>Ureaplasma urealyticum</i> (19,239 genomes/ml). Negative results for DNA samples from 3 different ubiquitous bacteria demonstrated the specificity of the PCR assay for <i>Mollicutes</i>. Direct testing of cell culture supernatants spiked with <i>Mycoplasma orale</i> revealed similar sensitivity compared to isolated DNA. <b><i>Conclusions: </i></b>Our single-tube real-time PCR assay with internal reaction control enables rapid and specific detection of mollicute contaminants. The test protocol is suitable for routine quality control of cell therapeutics." @default.
• W1996776500 created "2016-06-24" @default.
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• W1996776500 date "2013-12-19" @default.
• W1996776500 modified "2023-10-15" @default.
• W1996776500 title "A Single-Tube Real-Time PCR Assay for Mycoplasma Detection as a Routine Quality Control of Cell Therapeutics" @default.
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• W1996776500 doi "https://doi.org/10.1159/000357096" @default.
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