Diagnostic Uses of Monoclonal Antibodies to Salmonella

Lim, Pak Leong

doi:10.1016/b978-0-12-463003-1.50008-3

Cited by 5 publications

(7 citation statements)

References 91 publications

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“…Narrowing this antigen to the immunodominant O9 determinant would, in theory, increase the specificity of the assay. Indeed, using an inhibition ELISA to measure anti-O9 antibodies in patients, we observed very good specificity with the test previously (8,12). The high specificity resulting from measuring anti-O9 antibodies may explain why a healthy subject (subject N2) in the present study who had elevated levels of IgM and IgG anti-LPS antibodies and a high Widal TO had negative results in both the TUBEX test and the inhibition-ELISA.…”

Section: Discussionsupporting

confidence: 54%

“…The most common assay method used is the enzymelinked immunosorbent assay (ELISA), and the detecting antigen used in these assays is a purified extract of S. typhi cells. The antigen is derived from various subcellular structures of the organism, including the lipopolysaccharide (LPS), the outer membrane (OM), the flagella (d-H), and the capsule (virulence [Vi] antigen) (8). As expected, LPS-based (15,17,18,20,22) and OM-based (3,16,24) ELISAs were found to be superior to the Widal test.…”

mentioning

confidence: 74%

“…However, many of these bacteria are not invasive and may not stimulate a systemic antibody response. The extent to which TUBEX detects infection caused by these salmonellae or the paratyphoid serotypes (8,12) remains to be investigated. Previously, using the ELISA equivalent of TUBEX (12), we found that serum samples from septicemic patients infected with Salmonella organisms not belonging to serogroup D (one with S. choleraesuis, one with S. johannesburg, and one with S. senftenberg) were negative in the test, whereas that from a patient infected systemically with S. sendai (serogroup D) was weakly positive.…”

Section: Discussionmentioning

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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Section: Discussionsupporting

confidence: 54%

mentioning

confidence: 74%

Section: Discussionmentioning

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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“…Our laboratory has previously produced MAbs against Salmonella O antigen 9 (9). Together with the present MAb, which identifies O antigen 2, these specific serological reagents will allow the identification of the O antigens of S. typhi and S. paratyphi A, which are common causes of enteric fever.…”

Section: Resultsmentioning

confidence: 96%

“…However, this procedure is cumbersome, and batch-to-batch antiserum variations are also inevitable. Previously, our laboratory reported the production of monoclonal antibodies (MAbs) to the Salmonella LPS antigens (9,15), and we have demonstrated their usefulness in the serological identification of these organisms. We now report the production and characterization of a MAb, M02, raised against S. paratyphi A.…”

mentioning

confidence: 99%

Murine monoclonal antibody specific for lipopolysaccharide of Salmonella serogroup A

John.

Tsang

1987

J Clin Microbiol

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To facilitate the routine identification of salmonella and detailed studies of their lipopolysaccharides, we raised murine monoclonal antibodies against these organisms. We raised an immunoglobulin GI antibody, M02, which is specific for factor 02. By immunoblotting following sodium dodecyl sulfate-polyacrylamide gel electrophoresis, M02 was shown to bind only the lipopolysaccharide of Salmonella paratyphi A, giving a ladderlike reaction pattern with regularly spaced reactive bands. M02 did not react against lipopolysaccharides of other Salmonella serogroups, including those of serogroups B, C, D, E, and L. Since the lipopolysaccharides of Salmonella serogroups A, B, D, and E are similar except for the presence of paratose in serogroup A organisms, this dideoxyhexose is therefore believed to be the immunodominant epitope for M02. Consistent with the latter contention was the finding that periodate oxidation of the S. paratyphi A lipopolysaccharide did not destroy its antigenicity for M02. In a slide agglutination test, M02 was found to react specifically against al 12 clinical isolates of S. paratyphi A but not against 98 isolates of other salmonella or 74 isolates of other bacteria and Candida albicans.

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Molecular Structures of Bacteria Elucidated by Monoclonal Antibodies with Special Reference to Antigenic Determinants of the Methanogens' Envelopes

Macario

1986

Monoclonal Antibodies Against Bacteria

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Diagnostic Uses of Monoclonal Antibodies to Salmonella

Cited by 5 publications

References 91 publications

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

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

Murine monoclonal antibody specific for lipopolysaccharide of Salmonella serogroup A

Molecular Structures of Bacteria Elucidated by Monoclonal Antibodies with Special Reference to Antigenic Determinants of the Methanogens' Envelopes

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