Enzyme-linked immunoassay for detection of PCR-amplified DNA of legionellae in bronchoalveolar fluid.

Journal of Clinical Microbiology
D JonasS Bhakdi

Abstract

A nonradioactive method is described that detects 10 to 100 legionellae in 1 ml of bronchoalveolar lavage fluid. DNA is purified by a proteinase K-phenol protocol or with a commercial DNA preparation kit and amplified by PCR with amplimers specific for the 16S rRNA gene of Legionella pneumophila. The upstream primer is 5' biotinylated. The amplification product is immobilized on streptavidin-coated microtiter plates. Because of the high binding capacity, no removal of nonincorporated biotin from the PCR product is required. After alkaline denaturation, the single-stranded PCR product is hybridized with a 5' digoxigenin-labeled probing oligomer. The amplification product is then detected by using peroxidase-labeled anti-digoxigenin antibodies in a luminescence or colorimetric reaction. The assay detects as few as 10 legionellae in 1-ml bronchoalveolar lavage fluid specimens. It is specific for medically relevant Legionella species, including Legionella pneumophila, L. bozemanii, and L. longbeachae. Of over 250 clinical specimens examined, 8 were positive for legionellae by both culture and the PCR assay. Six further specimens were culture negative but PCR positive for legionellae; of these, five specimens were from patients rece...Continue Reading

References

Apr 1, 1992·Journal of Clinical Microbiology·B JaulhacH Monteil
Jun 1, 1990·Molecular and Cellular Probes·M H MahbubaniR M Atlas
Sep 1, 1990·Infection and Immunity·N P CianciottoN C Engleberg
Mar 1, 1986·Journal of Clinical Microbiology·P H Edelstein
Feb 1, 1986·Journal of Clinical Microbiology·H W WilkinsonB B Plikaytis
Oct 1, 1989·Journal of Clinical Microbiology·A W PasculleK Vrsalovic
Jun 1, 1989·Journal of Clinical Microbiology·M N StarnbachL S Tompkins
Aug 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·R K SaikiH A Erlich
Apr 1, 1989·Infection and Immunity·N C EnglebergB I Eisenstein
Jan 1, 1986·Nucleic Acids Research·E Huysmans, R De Wachter
Oct 1, 1984·Journal of Clinical Microbiology·R B KohlerL J Wheat
May 1, 1984·The Journal of Infectious Diseases·A L ReingoldJ N Kuritsky
Mar 1, 1994·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·G Lisby, R Dessau
Jun 1, 1994·Journal of Clinical Microbiology·P Matsiota-BernardC Nauciel

❮ Previous
Next ❯

Citations

Dec 20, 2002·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·David R Murdoch
Jan 11, 2005·Applied and Environmental Microbiology·Kathy B SheehanMichael J Ferris
Dec 16, 2004·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·J W Den Boer, E P F Yzerman
Mar 5, 2005·Médecine sciences : M/S·Catherine DaugaAbdelghani Sghir
Jan 13, 2012·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Vivian LuchsingerLuis F Avendaño
Apr 25, 2006·Journal of Applied Microbiology·S DutilC Duchaine
Mar 7, 2002·Journal of Clinical Microbiology·Anneke van der ZeeKate E Templeton
Dec 8, 2005·Applied and Environmental Microbiology·Steven GiglioChristopher P Saint
Sep 23, 2000·Infection Control and Hospital Epidemiology : the Official Journal of the Society of Hospital Epidemiologists of America·J BorauR A Venezia
Jan 11, 2005·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·David R Murdoch
Nov 18, 2009·New South Wales Public Health Bulletin·Christopher C BlythSharon C A Chen
Feb 26, 2013·Environmental Science and Pollution Research International·Alejandra Serrano-SuárezRosa Araujo
Sep 28, 2010·Acta Microbiologica Et Immunologica Hungarica·T FelföldiZ G Homonnay
Jul 1, 2014·Current Opinion in Infectious Diseases·Kyle R BrownbackSteven Q Simpson
Jan 16, 2014·Indian Journal of Microbiology·Cristina GruasM Victoria Arruga
May 10, 2007·Proceedings of the National Academy of Sciences of the United States of America·C D SinigallianoH M Solo-Gabriele
Sep 30, 2006·Scandinavian Journal of Infectious Diseases·Peter KorosecMitja Kosnik
Aug 30, 2001·Applied and Environmental Microbiology·N WellinghausenR Marre
Jul 5, 2002·Clinical Microbiology Reviews·Barry S FieldsRichard E Besser
Apr 5, 2003·Journal of Medical Microbiology·S AokiS Kamihira
Dec 19, 2008·Pakistan Journal of Biological Sciences : PJBS·Majid SoheiliMahmood Jeddi Tehrani
Apr 26, 2003·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·David R Murdoch
Apr 7, 2006·Applied and Environmental Microbiology·Philippe JolySophie Jarraud
Feb 15, 2018·The Journal of Eukaryotic Microbiology·Fatma GomaaColleen M Cavanaugh
Sep 28, 2001·Journal of Clinical Microbiology·B BöddinghausT Bittner
Jul 5, 2003·Journal of Clinical Microbiology·Deborah A WilsonGary W Procop
Aug 3, 2007·Journal of Clinical Microbiology·M A BenciniR Jansen

❮ Previous
Next ❯

Related Concepts

Related Feeds

CSF & Lymphatic System

This feed focuses on Cerebral Spinal Fluid (CSF) and the lymphatic system. Discover the latest papers using imaging techniques to track CSF outflow into the lymphatic system in animal models.

Related Papers

Pakistan Journal of Biological Sciences : PJBS
Majid SoheiliMahmood Jeddi Tehrani
Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases
Sverker BernanderLars-Victor Von Stedingk
© 2021 Meta ULC. All rights reserved