TY - THES A1 - Schiffers, Lukas T1 - Optimization of RT-PCR Assays for the Roche cobas® 6800/8800 Utility Channel: Influence of 2´-O-Methyl modified Primers on PCR efficiency N2 - Automated, high-throughput real-time PCR instruments such as the cobas® 6800/8800 from Roche have revolutionized the operation in molecular diagnostics laboratories as they provide a complete process of patient samples including all necessary steps for PCR applications from material preparation up to the calculation and evaluation of final results with a minimum of user interaction. In addition to the broad spectrum of commercially available ready-to-use reagent kits for blood screenings, virologic and microbiological assays as well as tests designed for women’s health, the cobas® 6800/8800 provides a tool which allows users to develop and implement their own tests onto the high-throughput device. Equipped with this tool, called the cobas® omni Utility Channel, these automated instruments serve a high flexibility and can be adjusted to users’ personal needs. Notwithstanding the advantages of the Utility Channel, a limited number of lab developed tests have been implemented onto the cobas® 6800/8800 so far as no guideline for the optimization and transfer of such tests onto the Utility Channel has been provided to the public yet. In the course of this bachelor thesis, four real-time PCR assays, including an allele-specific duplex test for HLA-B27, an established in-house test for the differentiation and detection of Ureaplasma urealyticum/parvum, a published assay for the detection of Chlamydia trachomatis as well as a duplex reverse-transcription assay for the detection and differentiation of norovirus genogroup one and two have been optimized for the use on Utility Channel. Furthermore, 2´-O-Methyl modified primers have been integrated into the optimization process of all assays to asses their influence on PCR efficiency. Three of these four assays have been transferred successfully onto a cobas® 6800 following a developed method which lead to the final design of a flowchart guideline for the optimization of lab developed tests for the use on a cobas® 6800/8800 system. KW - PCR efficiency KW - Modified primers KW - Methylated primers KW - Cobas KW - Molecular biology Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:hbz:1383-opus4-4632 ER -