The detection of Salmonella enteritidis and S. typhimurium using immunomagnetic separation and conductance microbiology

Parmar, Nimesh; Easter, M.C.; Forsythe, Stephen

doi:10.1111/j.1472-765x.1992.tb00756.x

Cited by 41 publications

(11 citation statements)

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“…Parmar et al (20) evaluated an IMS-conductance method for detection of Salmonella in milk powders. In evaluating the specificity of IMS by using anti-Salmonella Dynabeads, they noted that exposure to Citrobacter freundii resulted in a Salmonella type of conductance curve and concluded that the magnetic beads were only partially specific for the Salmonella strains tested.…”

Section: Discussionmentioning

confidence: 99%

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Development and Optimization of a Novel Immunomagnetic Separation- Bacteriophage Assay for Detection of Salmonella enterica Serovar Enteritidis in Broth

Favrin¹,

Jassim²,

Griffiths

2001

Appl Environ Microbiol

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Salmonella is the second-leading cause of food-borne illness in most developed countries, causing diarrhea, cramps, vomiting, and often fever. Many rapid methods are available for detection of Salmonella in foods, but these methods are often insensitive or expensive or require a high degree of technical ability to perform. In this paper we describe development and characterization of a novel assay that utilizes the normal infection cycle of bacteriophage SJ2 for detection of Salmonella enterica serovar Enteritidis in broth. The assay consists of four main stages: (i) capture and concentration of target cells by using immunomagnetic separation (IMS); (ii) infection of the target bacterium with phage; (iii) amplification and recovery of progeny phage; and (iv) assay of progeny phage on the basis of their effect on a healthy population of host cells (signal-amplifying cells). The end point of the assay can be determined by using either fluorescence or optical density measurements. The detection limit of the assay in broth is less than 10 4 CFU/ml, and the assay can be performed in 4 to 5 h. The results of this study demonstrate that the IMS-bacteriophage assay is a rapid, simple, and sensitive technique for detection of Salmonella serovar Enteritidis in broth cultures which can be applied to preenriched food samples.

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Section: Discussionmentioning

confidence: 99%

“…IMS can eliminate the need for selective enrichment and reduce the time required for conventional methods by as much as 24 h (15). IMS has also been used in conjunction with other rapid detection methods, including enzyme-linked immunosorbent assays (ELISAs), conductance microbiology, electrochemiluminescence, and PCR (6,8,12,20,34).…”

mentioning

confidence: 99%

Development and Optimization of a Novel Immunomagnetic Separation- Bacteriophage Assay for Detection of Salmonella enterica Serovar Enteritidis in Broth

Favrin¹,

Jassim²,

Griffiths

2001

Appl Environ Microbiol

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“…In addition to standard bacteriological culture, detection methods reported for salmonella include ELISA, utilizing monoclonal antibodies [25,26], fluorogenic detection using the enzyme substrate 4-methylumbrelliferylcaprylate (MUCAP) [27,28], a latex slide agglutination test (Mercia Diagnostics) and a conductance test [29,30]. These tests are quicker than the conventional bacteriological method but they are neither quantitative nor sensitive enough to detect small numbers of bacteria [31].…”

Section: Discussionmentioning

confidence: 99%

A quantitative polymerase chain reaction method for the detection in avian faeces of salmonellas carrying thespvRgene

Mahon¹,

Lax²

1993

Epidemiol. Infect.

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SummaryA quick, semi-quantitative method of detectingSalmonellaspecies which contain the virulence plasmid has been developed using the polymerase chain reaction (PCR). A pair of primers have been synthesized encompassing a 500 bp fragment of thespvRvirulence gene. Competitor DNA consisting of thespvRgene with a 94 bp deletion situated between the primer recognition sequences, was cloned into a plasmid vector. Co-amplification of the ‘unknown’ target salmonella DNA with known quantities of competitor DNA in the same reaction tube gave PCR products of 500 and 406 bp respectively. Visual assessment of the ratio of the two products on ethidium bromide stained agarose gels provided an estimate of the approximate number of salmonella cells present in avian faeces.The technique could be applied to detect quantifiably any non-host DNA in clinical samples if a suitable DNA sequence for primer construction is available.

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Section: Dot Immunoblotting (Dib) Procedurementioning

confidence: 99%

“…Percentage data were subjected to arcsine transformation (Sokal & Rohlf, 1969), prior to analysis of variance using the MINITAB statistical package (Minitab Statistical Systems). Mean significant difference (MSD) values, at the 95% confidence level, were calculated from the analysis of variance (Petersen, 1985).Isolation of target by IMC. Antibody-labelled spores were captured by sheep anti-rabbit (SAR) M-280 Dynabeads (Dynal, UK) and recovered with a magnetic particle concentrator (MPC), product numbers 12002 and 12004 (Dynal, UK).…”

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confidence: 99%

Selective recovery of Streptosporangium fragile from soil by indirect immunomagnetic capture

Mullins

Gürtler²,

Wellington³

1995

Microbiology

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A polyclonal anti body raised to Stmptosporangium fkagi/e spores reacted strongly and specifically with the immunizing strain and to a number of related species of Streptosporangium, as determined by dot immunoblotting. An indirect immunomagnetic capture method was developed for the recovery of the target organism from sterile and non-sterile soil, using sheep anti-rabbit M-280 Dynabeads. The effects of different soil blocking agents, antibody labelling concentrations and spore/Dynabead capture times on the recovery of 5. fragile spores were investigated. Pre-blocking of antibody binding sites within the soil, with either 2% partially hydrolysed gelatin or 10% skimmed milk, was essential prior to immunomagnetic capture. Increasing the capture time from 15 to 60 min did not affect spore recovery; however, a 10-fold decline in the magnetic bead concentration did result in a significantly lower recovery of spores from soil. 5. fragi/e was selectively enriched (1 :190=fold) when present as a mixed population with Arthmbacter oxydans in sterile soil. The indirect immunomagnetic capture method was used to selectively recover 5. fragile spores seeded into non-sterile soil, although some background binding of non-target bacteria was noted. The target was successfully recovered from a sterile soil microcosm after 14 d incubation and the capture rate was increased by the inclusion of an initial soil dispersion and biomass concentration procedure, using the ion-exchange resin Chelex 100.Keywords : immunocapture, Streptosporangium, soil, magnetic beads, selective isolation INTRODUCTIONThere is an increasing need to improve methods for the detection and isolation of bacteria from environmental samples, both for the recovery of genetically engineered strains and microbial inoculants (Ford & Olsen, 1988;Morgan et al., 1989; Trevors e t al., 1993) and for the isolation of rare and unusual bacteria for use in industrial screening programmes (Nolan & Cross, 1988 ;Shearer, 1987).Immunomagnetic capture (IMC) represents a potentially important development within this field, allowing the specific recovery of target bacteria from a diverse range of environmental samples. The method relies upon the interaction between cell-surface antigens and antibodies which can be attached to magnetized polystyrene beads.Abbreviations: DIB, dot irnrnunoblotting; IMC, irnrnunornagnetic capture; MPC, magnetic partlcle concentrator; Non, Nonidet P-40.Separation of target cells from a suspension can be achieved using a magnetic field. IMC has been widely used in clinical studies for the separation or depletion of target cells from a mixed population (Gaudernack et ai., 1986; Gruhn e t al., 1991 ; Silvestri e t al., 1992). More recently it has been used to enrich for and enhance the detection of pathogenic bacteria present in contaminated foodstuffs (Johne e t al., 1989; Mansfield e t a!., 1993; Parmer eta/., 1992; Skjerve etai., 1990) and faeces (Luk & Lindberg, 1991 ; Tomoyasu, 1992). Work has also focused on the detection and isolation of bacteri...

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The detection of Salmonella enteritidis and S. typhimurium using immunomagnetic separation and conductance microbiology

Cited by 41 publications

References 5 publications

Development and Optimization of a Novel Immunomagnetic Separation- Bacteriophage Assay for Detection of Salmonella enterica Serovar Enteritidis in Broth

Development and Optimization of a Novel Immunomagnetic Separation- Bacteriophage Assay for Detection of Salmonella enterica Serovar Enteritidis in Broth

A quantitative polymerase chain reaction method for the detection in avian faeces of salmonellas carrying thespvRgene

Selective recovery of Streptosporangium fragile from soil by indirect immunomagnetic capture

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