Wenzel, Juergen J. and Walch, Heiko and Bollwein, Markus and Niller, Hans Helmut and Ankenbauer, Waltraud and Mauritz, Ralf and Hoeltke, Hans-Joachim and Manuel Zepeda, Hector and Wolf, Hans and Jilg, Wolfgang and Reischl, Udo (2009) Library of Prefabricated Locked Nucleic Acid Hydrolysis Probes Facilitates Rapid Development of Reverse-Transcription Quantitative Real-Time PCR Assays for Detection of Novel Influenza A/H1N1/09Virus. CLINICAL CHEMISTRY, 55 (12). pp. 2218-2222. ISSN 0009-9147,
Full text not available from this repository. (Request a copy)Abstract
BACKGROUND: The emergence of a novel pandemic human strain of influenza A (H1N1/09) has clearly demonstrated the need for flexible tools enabling the rapid development of new diagnostic methods. METHODS: We designed a set of reverse-transcription quantitative real-time PCR (RT-qPCR) assays based on the Universal ProbeLibrary (UPL)-a collection of 165 presynthesized, fluorescence-labeled locked nucleic acid (LNA) hydrolysis probes-specifically to detect the novel influenza A virus. We evaluated candidate primer/UPL-probe pairs with 28 novel influenza A/H1N1/09 patient samples of European and Mexican origin. RESULTS: Of 14 assays in the hemagglutinin (HA) and neuraminidase (NA) genes, 12 detected viral nucleic acids from diluted patient samples without need for further optimization. We characterized the diagnostic specificity of the 2 best-performing assays with a set of samples comprising various influenza virus strains of human and animal origin that showed no cross-reactivity. The diagnostic sensitivity of these 2 primer/probe combinations was in the range of 100-1000 genomic copies/mL. In comparison to a reference assay recommended by the German health authorities, the analytical sensitivities and specificities of the assays were equivalent. CONCLUSIONS: Facing the emergence of novel influenza A/H1N1/09, we were able to develop, within 2 days, a set of sensitive and specific RT-qPCR assays for the laboratory diagnosis of suspected cases. H1N1/09 served as a model to show the feasibility of the UPL approach for the expedited development of new diagnostic assays to detect emerging pathogens.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | A H1N1 VIRUS; RT-PCR; INFECTION; EMERGENCE; PANDEMICS; LNA; |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 27 Aug 2020 12:30 |
| Last Modified: | 27 Aug 2020 12:30 |
| URI: | https://pred.uni-regensburg.de/id/eprint/28006 |
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