Sandwich-Dot Immunobinding Assay (Sandwich-DIA), a New Immunological Method for the Detection of Diphtheria Toxin

Pietrzak, J.; Muehlestein, S.; Gasser, Markus

doi:10.1016/s0934-8840(11)80975-8

Cited by 7 publications

(10 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This limit of detection is comparable to that of the amplified EIA that we described previously (limit of detection, 0.1 ng/ml) (7), is 20-fold more sensitive than other EIAs (limit of detection, 10 ng/ml) (14,16), and is 10-fold more sensitive than agglutination assays (12,17).…”

Section: Discussionsupporting

confidence: 80%

“…The phenotypic methods available for the detection of diphtheria toxin tend to be technically demanding, lacking in sensitivity, and relatively expensive or have not been fully evaluated against a large panel of isolates (5,8,10,12,14,16,17). We recently described an amplified EIA for the detection of diphtheria toxin (7) which had a number of advantages over previously documented phenotypic methods for the detection of toxigenicity, including improved sensitivity, specificity, and speed.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Immunochromatographic Strip Test for Rapid Detection of Diphtheria Toxin: Description and Multicenter Evaluation in Areas of Low and High Prevalence of Diphtheria

et al. 2002

View full text Add to dashboard Cite

An immunochromatographic strip (ICS) test was developed for the detection of diphtheria toxin by using an equine polyclonal antibody as the capture antibody and colloidal gold-labeled monoclonal antibodies specific for fragment A of the diphtheria toxin molecule as the detection antibody. The ICS test has been fully optimized for the detection of toxin from bacterial cultures; the limit of detection was approximately 0.5 ng of diphtheria toxin per ml within 10 min. In a comparative study with 915 pure clinical isolates of Corynebacterium spp., the results of the ICS test were in complete agreement with those of the conventional Elek test. The ICS test was also evaluated for its ability to detect toxigenicity from clinical specimens (throat swabs) in two field studies conducted within areas of the former USSR where diphtheria is epidemic. Eight hundred fifty throat swabs were examined by conventional culture and by use of directly inoculated broth cultures for the ICS test. The results showed 99% concordance (848 of 850 specimens), and the sensitivity and specificity of the ICS test were 98% (95% confidence interval, 91 to 99%) and 99% (95% confidence interval, 99 to 100%), respectively.

show abstract

Section: Discussionsupporting

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Immunochromatographic Strip Test for Rapid Detection of Diphtheria Toxin: Description and Multicenter Evaluation in Areas of Low and High Prevalence of Diphtheria

et al. 2002

View full text Add to dashboard Cite

show abstract

“…Other immunological assays Several immunological approaches including agglutination (Jalgaonkar and Saoji 1993;Maximescu and Fîciu 1980;Toma et al 1997), counterimmunoelectrophoresis (Thompson and Ellner 1978), enzyme immunoassays (Engler and Efstratiou 2000;Hallas et al 1990;Krech and Wittelsbürger 1987;Nielsen et al 1987) or dot immunobinding methods (Pietrzak et al 1990) have been chosen to detect DT from Corynebacterium isolates or clinical samples. However, most of these assays are technically demanding, lack sensitivity, are relatively expensive, were only published in non-English journals (mainly in Russian and Romanian) or have been evaluated only with a very small panel of isolates.…”

Section: The Immunochromatographic Strip Test (Ics)mentioning

confidence: 99%

Detection Methods for Laboratory Diagnosis of Diphtheria

Berger

Hogardt

Konrad

et al. 2013

Corynebacterium Diphtheriae and Related Toxigenic Species

View full text Add to dashboard Cite

Although diphtheria has to be diagnosed primarily on clinical symptoms, the rapid and reliable detection and identification of the potentially toxigenic Corynebacterium species, C. diphtheriae, C. ulcerans and C. pseudotuberculosis, is essential for the definite diagnosis and management of diphtheria with respect to both the individual patient and the public health measures to be undertaken. Laboratory confirmation of suspected diphtheria has to aim for the isolation of the etiologic pathogen (including species identification and antibiotic susceptibility testing) as well as for differentiation of toxigenic from non-toxigenic strains (by using tox gene detection and toxigenicity testing). The recent introduction of the Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) into the microbiological algorithm of laboratory diagnosis of diphtheria allows the specific and escalating identification of the three potentially toxigenic Corynebacterium species from growing colonies. Finally, molecular typing techniques may be applied to explore the clonal relatedness of clinical isolates and their potential routes of transmission. The most relevant laboratory procedures fulfilling these requirements (microbiological culture, conventional biochemical tests, molecular methods for species identification and toxigenicity testing) will be presented here.

show abstract

“…EIAs for the detection of diphtheria toxin have been previously documented (13,23,27). The majority of these assays used similar designs (horse polyclonal antitoxin as the capture antibody and a mouse monoclonal antibody as the detecting antibody), with the exception of that of Hallas et al (13), who used two monoclonal antibodies specific to fragment A as both the detecting and capture antibodies.…”

Section: Discussionmentioning

confidence: 99%

“…Although genotypic PCR-based methods for the detection of the toxin gene (21,22,24) offer some advantages over phenotypic tests, they do not provide information on the ability of the organism to express biologically active diphtheria toxin and therefore cannot provide a definitive result on toxigenicity (7,25). Enzyme immunoassays (EIAs) are widely used for the detection of microbial antigens and markers (13,23,27). The sensitivity of two-site immunometric EIAs can be improved by the incorporation of signal amplification technology (2,20).…”

mentioning

confidence: 99%

Rapid Enzyme Immunoassay for Determination of Toxigenicity among Clinical Isolates of Corynebacteria

Engler

Efstratiou

2000

J Clin Microbiol

View full text Add to dashboard Cite

A rapid enzyme immunoassay (EIA) was developed for the phenotypic detection of diphtheria toxin among clinical isolates of corynebacteria. The assay uses equine polyclonal antitoxin as the capture antibody and an alkaline phosphatase-labeled monoclonal antibody, specific for fragment A of the toxin molecule, as the detecting antibody. The assay is rapid, sensitive, and specific: a final result is available within 3 h of colony selection, and the limits of detection are 0.1 ng of pure diphtheria toxin/ml. Toxigenicity could be detected with isolates grown on a diverse range of culture media, including selective agars. Toxin detection using the EIA was compared to that with the Elek test and PCR detection of fragment A of the diphtheria toxin (tox) gene, using 245 isolates of corynebacteria. The results for the EIA were in complete concordance with those of the Elek test: 87 toxigenic and 158 nontoxigenic isolates. Ten of the phenotypically nontoxigenic strains were found to contain fragment A of the tox gene but did not express the toxin protein. These isolates were found to be nontoxigenic in the Vero cell tissue culture cytotoxicity assay and were therefore nontoxigenic for diagnostic purposes. The EIA is a simple rapid phenotypic test which provides a definitive result on toxigenicity within one working day.

show abstract

Sandwich-Dot Immunobinding Assay (Sandwich-DIA), a New Immunological Method for the Detection of Diphtheria Toxin

Cited by 7 publications

References 20 publications

Immunochromatographic Strip Test for Rapid Detection of Diphtheria Toxin: Description and Multicenter Evaluation in Areas of Low and High Prevalence of Diphtheria

Immunochromatographic Strip Test for Rapid Detection of Diphtheria Toxin: Description and Multicenter Evaluation in Areas of Low and High Prevalence of Diphtheria

Detection Methods for Laboratory Diagnosis of Diphtheria

Rapid Enzyme Immunoassay for Determination of Toxigenicity among Clinical Isolates of Corynebacteria

Contact Info

Product

Resources

About