Kern, Monika and Boehm, Stephan and Deml, Ludwig and Wolf, Hans and Reischl, Udo and Niller, Hans Helmut (2009) Inhibition of Legionella pneumophila PCR in respiratory samples: A quantitative approach. JOURNAL OF MICROBIOLOGICAL METHODS, 79 (2). pp. 189-193. ISSN 0167-7012, 1872-8359
Full text not available from this repository. (Request a copy)Abstract
Impurities in complex biological samples that persist through nucleic acid preparation can inhibit PCR or reduce the sensitivity and efficiency of PCR amplification and thus affect reliable results in PCR diagnostics. To obtain information on the incidence of inhibition events and on the magnitude of the loss of sensitivity, respectively, we devised a relative inhibition assay and examined 100 samples from patients with respiratory tract infections. As a reference, samples were spiked with Legionella (L) pneumophila. Detection was by standard nucleic acid purification and subsequent real-time PCR. By comparing the crossing points of the fluorescent curves to those of an L pneumophila standard dilution series, we were able to quantify the respective degrees of inhibition into several categories. We found complete inhibition in 2% of the samples. 12% were not reliably detected. 65% of the tested samples showed moderate to strong inhibition, but were still reliably detected, whereas in 21% of the samples no inhibition was observed. Except for a significantly higher inhibition in tracheal aspirates than in BAL samples, the degree of inhibition did not correlate with the physical properties of the respective sample. The relative inhibition assay established an unexpectedly broad distribution of the inhibition-degrees in inflammatory respiratory materials. (C) 2009 Elsevier B.V. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | POLYMERASE-CHAIN-REACTION; MYCOBACTERIUM-TUBERCULOSIS COMPLEX; BRONCHOALVEOLAR LAVAGE FLUIDS; REVERSE TRANSCRIPTION-PCR; NUCLEIC-ACID EXTRACTION; RNA INTERNAL CONTROL; HUMAN FECAL SAMPLES; REAL-TIME PCR; CHLAMYDIA-PNEUMONIAE; CLINICAL SPECIMENS; Real-time PCR; Crossing point; Partial inhibition; Legionella; Respiratory samples |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 02 Sep 2020 10:06 |
| Last Modified: | 02 Sep 2020 10:06 |
| URI: | https://pred.uni-regensburg.de/id/eprint/28212 |
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