Detection of group D salmonellae in blood culture broth and of soluble antigen by tube agglutination using an O-9 monoclonal antibody latex conjugate

Lim, Pak Leong; Fok, Y P

doi:10.1128/jcm.25.7.1165-1168.1987

Cited by 26 publications

(8 citation statements)

References 14 publications

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“…Recently, we established an MAb-based immunomagnetic tech-nique to open a new possibility for the rapid detection of salmonellae in clinical and food specimens (23). Furthermore, there have been several reports on the use of MAb probes to detect salmonellae in clinical specimens, and their potential diagnostic value was also demonstrated (3,19,30,35), even though either the epitope specificity of the described MAbs were not immunochemically defined or the variety of salmonellae identified by these agents was often limited. With a satisfactory performance in slide agglutination tests, the present 0-antigen-specific MAbs correctly identified all 167 strains of salmonellae from serogroups B to E (Table 6).…”

Section: Discussionmentioning

confidence: 99%

Anti-Salmonella lipopolysaccharide monoclonal antibodies: characterization of Salmonella BO-, CO-, DO-, and EO-specific clones and their diagnostic usefulness

Luk

Lindberg

1991

J Clin Microbiol

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To facilitate the identification and serotyping of Salmonella species, we established a wide variety of murine monoclonal antibodies (MAbs) that were reactive with the lipopolysaccharides (LPSs) of Salmonella serogroups B to E. An effective approach for generating LPS-reactive hybridomas was used; this required immunization of mice with LPS-coated bacteria. To screen for diagnostically useful MAbs, the MAbs were tested by enzyme-linked immunosorbent assay against a set of purified LPSs from smooth and rough Salmonella strains. At least four major groups of antibody specificities were identified: Salmonella (i) BO specific, (ii) CO specific, (iii) DO specific, and (iv) E0 specific. For a more detailed epitope analysis, a panel of eight different serogroup-specific MAbs which were shown to bind the 0-antigenic polysaccharide chains, yielding characteristic ladder patterns in Western blots (immunoblots) against the LPS of Salmonella serogroups B to E, were selected. The availability of various chemically defined LPS structures and Salmonella 0-antigen glycoconjugates permitted the definition of 0-antigenic polysaccharide epitopes recognized by each MAb that serologically corresponded to factors 03, 04, 05, 06, 07, 08, 09, and 010 on the basis of the Kauffmann-White scheme for Salmonella classification. The diagnostic accuracy of these immunochemically defined 0-specific MAbs for Salmonella serotyping was demonstrated by correct identification of all 167 salmonellae (including 72 serotypes from serogroups B to E) among the 294 bacterial strains in a slide agglutination test. No false-positive reactions were detected.

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Section: Discussionmentioning

confidence: 99%

Anti-Salmonella lipopolysaccharide monoclonal antibodies: characterization of Salmonella BO-, CO-, DO-, and EO-specific clones and their diagnostic usefulness

Luk

Lindberg

1991

J Clin Microbiol

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“…previously (12). The antibody was produced in mouse ascitic fluid and purified by cryoprecipitation (11). The immunoglobulin content of the purified preparation was determined by single radial immunodiffusion (5) using goat anti-serum (Sigma Chemical Co., St. Louis, Mo.)…”

Section: Methodsmentioning

confidence: 99%

“…Particle conjugation. White latex particles (diameter, 0.8 m; Sigma) were sensitized with purified anti-O9 antibody or S. typhi LPS (Difco Laboratories, Detroit, Mich.) by passive adsorption as described previously (11).…”

Section: Methodsmentioning

confidence: 99%

“…2, the set comprises six identical V-shaped tubes, each with a capacity of 1 ml. For reagent A, magnetic particles (Dynal, Oslo, Norway) were coupled with S. typhi LPS (Difco) by passive adsorption as described previously (11). The particles were suspended as 0.25% (wt/vol) solids in 0.1 M glycine-borate buffer (pH 8.2) containing 0.9% NaCl (GBS buffer) and 1% BSA (GBS-BSA buffer).…”

Section: Methodsmentioning

confidence: 99%

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One-Step 2-Minute Test To Detect Typhoid-Specific Antibodies Based on Particle Separation in Tubes

et al. 1998

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Typhoid fever is caused by Salmonella typhi. Detection of anti-S. typhi antibodies in the patient is a useful diagnostic aid. Among the various methods developed over the years for this purpose, the Widal test, based on bacterial agglutination, has remained the most widely used, even though it is neither specific nor sensitive. Its popularity stems from the fact that it is simple to use and inexpensive. We describe a new test which also uses a simple one-step procedure but is more rapid and accurate than the Widal. The new test (TUBEX) detects anti-Salmonella O9 (both immunoglobulin M [IgM] and IgG) antibodies in patients by inhibiting the binding between an anti-O9 IgM monoclonal antibody (MAb) conjugated to colored latex particles and S. typhilipopolysaccharide (LPS) conjugated to magnetic latex particles. The reactants are mixed in a specially designed microtube for 2 min, and the result is read based on the resultant color of the supernatant following forced sedimentation of the magnetic beads. In the absence of inhibitory antibodies, there is a color change (from blue to red) due to cosedimentation of the indicator particles with the magnetic particles, whereas if these antibodies are present, they prevent such a change to a degree dependent on their concentration. Preliminary examination of TUBEX using the anti-O9 MAb and irrelevant MAbs as inhibitors revealed the test to be specific and reproducible, with an analytical sensitivity of 16 μg per ml of antibody. The reagents remained stable for at least 9 months when kept at 4°C. In the examination of 16 stored sera obtained from 14 patients with proven cases of typhoid fever and 78 serum samples from 75 subjects without typhoid fever, TUBEX was found to be 100% sensitive and 100% specific. The nontyphoid group comprised 26 healthy blood donors, 30 antinuclear antibody (ANA)-negative patients, 9 ANA-positive patients, of whom 1 was positive for anti-DNA antibody, 4 typhus patients, and 6 septicemic patients. In addition, the sera obtained from 11 patients clinically diagnosed as having typhoid fever were all positive in the test. The TUBEX results correlated to some extent, albeit insignificantly (r = 0.38,P = 0.07), with those of an enzyme-linked immunoassay (ELISA) which used a similar detection format (inhibition) and reagents (S. typhi LPS and anti-O9 antibody). TUBEX correlated very well with ELISAs which detected anti-S. typhi LPS IgM (r = 0.58, P = 0.003) or IgG (r = 0.54, P = 0.006) antibodies from the typhoid patients. There was no correlation with the Widal test. The TUBEX test, if performed on slides (instead of tubes) or with soluble antigen (instead of antigen-conjugated magnetic beads), suffered significantly in sensitivity. Direct agglutination tests using LPS-conjugated indicator particles performed either on slides or in microwells also failed to detect antibodies from the majority of typhoid patients. Thus, TUBEX appears to be well designed and well suited for use in the laboratory or by the bedside as a simple, rapid aid to the routine diagnosis of typhoid fever.

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“…The bacterial agglutination test (high titre serum agglutination test) was adopted to identify S. typhi (Lim and Fok 1987), S. paratyphi A and S. paratyphi B. This procedure was carried out on an ordinary glass slide.…”

Section: Serological Test For the Confirmation Of Salmonella Sppmentioning

confidence: 99%

An epidemiological surveillance of asymptomatic typhoid carriers associated in respect to socioeconomic status in India

Balakrishnan¹,

Duraisamy²,

Rajasekarapandian³

2012

J Public Health

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Detection of group D salmonellae in blood culture broth and of soluble antigen by tube agglutination using an O-9 monoclonal antibody latex conjugate

Cited by 26 publications

References 14 publications

Anti-Salmonella lipopolysaccharide monoclonal antibodies: characterization of Salmonella BO-, CO-, DO-, and EO-specific clones and their diagnostic usefulness

Anti-Salmonella lipopolysaccharide monoclonal antibodies: characterization of Salmonella BO-, CO-, DO-, and EO-specific clones and their diagnostic usefulness

One-Step 2-Minute Test To Detect Typhoid-Specific Antibodies Based on Particle Separation in Tubes

An epidemiological surveillance of asymptomatic typhoid carriers associated in respect to socioeconomic status in India

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