Rapid identification of legionellae by a colony blot assay based on a genus-specific monoclonal antibody

Steinmetz, Ivo; Rheinheimer, Claudia; Bitter‐Suermann, D.

doi:10.1128/jcm.30.4.1016-1018.1992

Cited by 17 publications

(5 citation statements)

References 5 publications

(5 reference statements)

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“…After the initial decrease, the number of G4 5223-PR1 cells remained stable for at least 25 days, indicating that starvation was not a primary factor in decreases in the numbers of G4 5223-PR1 cells in the systems which we studied. Direct immunofluorescent counting of G4 5223-PR1 cells in samples extracted from sediment was limited by non- The colony immunoblot assay, in which antibodies specific for peripheral antigens are used, is a rapid method which has been used previously to identify pathogenic microorganisms (2,34). This technique was used to detect G4 5223-PR1 in sediment samples, because it was not always possible to identify all G4 5223-PR1 colonies by the selective plating technique (Fig.…”

Section: Discussionmentioning

confidence: 99%

Tracking the Response of Burkholderia cepacia G4 5223-PR1 in Aquifer Microcosms

Winkler¹,

Timmis²,

Snyder³

1995

Appl Environ Microbiol

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The introduction of bacteria into the environment for bioremediation purposes (bioaugmentation) requires analysis and monitoring of microbial population dynamics to define persistence and activity from both efficacy and risk assessment perspectives. Burkholderia cepacia G4 5223-PR1 is a Tn5 insertion mutant which constitutively expresses a toluene ortho-monooxygenase that degrades trichloroethylene (TCE). This ability of G4 5223-PR1 to degrade TCE without aromatic induction may be useful for bioremediation of TCE-containing aquifers and groundwater. Thus, a simulated aquifer sediment system and groundwater microcosms were used to monitor the survival of G4 5223-PR1. The fate of G4 5223-PR1 in sediment was monitored by indirect immunofluorescence microscopy, a colony blot assay, and growth on selective medium. G4 5223-PR1 was detected immunologically by using a highly specific monoclonal antibody which reacted against the O-specific polysaccharide chain of the lipopolysaccharides of this organism. G4 5223-PR1 survived well in sterilized groundwater, although in nonsterile groundwater microcosms rapid decreases in the G4 5223-PR1 cell population were observed. Ten days after inoculation no G4 5223-PR1 cells could be detected by selective plating or immunofluorescence. G4 5223-PR1 survival was greater in a nonsterile aquifer sediment microcosm, although after 22 days of elution the number of G4 5223-PR1 cells was low. Our results demonstrate the utility of monoclonal antibody tracking methods and the importance of biotic interactions in determining the persistence of introduced microorganisms.

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Section: Discussionmentioning

confidence: 99%

Tracking the Response of Burkholderia cepacia G4 5223-PR1 in Aquifer Microcosms

Winkler¹,

Timmis²,

Snyder³

1995

Appl Environ Microbiol

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“…The nitrocellulose filters were developed with 4‐chloro‐1‐naphthol as the substrate. The reaction was stopped by rinsing the filters in tap water (Steinmetz and others 1992).…”

Section: Methodsmentioning

confidence: 99%

Rapid identification of porcine Serpulina species by colony blot assay using a genus‐specific monoclonal antibody

Mittal¹

1996

Veterinary Record

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An immunoglobulin G monoclonal antibody (mAb C9E8) recognising a genus-specific epitope on the 26 kDa protein of porcine Serpulina species organisms was used in a simple colony blot assay to detect Serpulina in cultures grown directly on blood agar plates from pig faeces and tissues. The mAb detected even a few colonies of the organism in the presence of an abundant growth of non-Serpulina organisms. The whole procedure was completed in less than three hours. A total of 123 strains of S hyodysenteriae and S innocens were correctly identified by the colony blot assay whereas all the 26 non-Serpulina Gram-negative organisms commonly isolated from faecal material or tissues of pigs remained negative. The assay was rapid, highly specific and sufficiently reliable to be used with confidence for identifying porcine Serpulina colonies directly on blood agar plates.

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“…The most important technique for the identification of legionellae in the clinical laboratory is the serological characterisation of isolated strains. Two genus-specific monoclonal antibodies (MAbs) have been described [15,16]. The advantage of such reagents is that the identification of the genus, which is of clinical significance, can be obtained with a single reagent.…”

Section: Culture and Identification Of Legionellaceaementioning

confidence: 99%

“…Steinmetz et al used a colony blot immunoassay, using a MAb against the 58±60 kDa heat-shock protein. The genus-specific MAb against the surface-exposed macrophage infectivity potentiator (Mip) protein can be detected in a whole cell enzyme-linked immunosorbent assay (ELISA) [15]. A fluorescein-conjugated MAb which recognises an outer membrane protein of L. pneumophila [17] is commercially available.…”

Section: Culture and Identification Of Legionellaceaementioning

confidence: 99%

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Epidemiology and Laboratory Diagnosis of Legionella Infections. Epidemiologie und Labordiagnose von Legionella-Infektionen

Lück

Helbig

Schuppler

2002

Laboratoriums Mediz

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Summary: After transfer from the environment, Legionella species may cause pneumonias or influenza‐like respiratory infections. The diagnostic methods currently available, such as culture, serology, direct fluorescent antibody testing, and urinary antigen detection, still constitute the basic diagnostic repertoire. However, none of the diagnostic tests presently available offers the desired quality with respect to sensitivity and specificity. Therefore, the standard performance is to use several diagnostic tests in parallel. Culture should be obligatory especially when hospitalized patients with underlying diseases are investigated. Urinary antigen detection is a valuable tool when L. pneumophila serogroup 1 is the causative agent. This is the case in the majority of community acquired cases. For the surveillance of nosocomial legionellosis, direct fluorescence antigen detection may be of greater value since this test detects all serogroups of L. pneumophila. The detection of antibodies in patient's sera is still a good laboratory test but is of little use in the acute phase of illness. Recently significant advances have been made in the development of diagnostic tests which are based on the detection of nucleic acids, most notably the polymerase chain reaction.

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Rapid identification of legionellae by a colony blot assay based on a genus-specific monoclonal antibody

Cited by 17 publications

References 5 publications

Tracking the Response of Burkholderia cepacia G4 5223-PR1 in Aquifer Microcosms

Tracking the Response of Burkholderia cepacia G4 5223-PR1 in Aquifer Microcosms

Rapid identification of porcine Serpulina species by colony blot assay using a genus‐specific monoclonal antibody

Epidemiology and Laboratory Diagnosis of Legionella Infections. Epidemiologie und Labordiagnose von Legionella-Infektionen

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