Serodiagnosis of typhoid fever by enzyme-linked immunosorbent assay determination of anti-Salmonella typhi lipopolysaccharide antibodies

Nardiello, S; Pizzella, T; Russo, Massimo; Galanti, B

doi:10.1128/jcm.20.4.718-721.1984

Cited by 43 publications

(8 citation statements)

References 4 publications

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“…In recent years more specific and sensitive techniques than agglutination, such as radioimmunoassay and enzymelinked immunosorbent assay (ELISA), have been used for detecting and quantitating the antibody levels primarily to Salmonella sp. 0 antigens (1,5,22,28). Although attempts have been made to characterize the humoral response to protein antigens other than the flagellar component, these have been performed either with poorly defined antigens or with mixed-antigen preparations (1,26,28).…”

mentioning

confidence: 99%

Antibodies to porin antigens of Salmonella typhi induced during typhoid infection in humans

Calderón¹,

Lobos²,

Rojas³

et al. 1986

Infect Immun

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mentioning

confidence: 99%

Antibodies to porin antigens of Salmonella typhi induced during typhoid infection in humans

Calderón¹,

Lobos²,

Rojas³

et al. 1986

Infect Immun

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“…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. The chemical structure of Salmonella LPS is, in fact, well-known (reviewed in references 6 and 8).…”

mentioning

confidence: 73%

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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“…A number of attempts have been made in the past to develop a reliable serological technique for the diagnosis of typhoid fever (Duthie and French, 1990). ese include the Widal agglutination test, haemagglutination test, enzyme-linked immunosorbent assay, immunoelectrophoresis and polymerase chain reaction test (Rai et al, 1989;Barrett et al, 1983;Nardiello et al, 1984;Verdugo-Rodriguez et al, 1993;Jesudason et al, 1998;Chart et al, 1997;Sharma et al, 1997). None of these tests could be adopted for routine use on account of various reasons whereby the Widal tube agglutination test remained a simple, inexpensive, semiquantitative (Parry et al, 1999;Choo et al, 1993;Saha et al, 1996;Coovadia et al, 1986).…”

Section: Introductionmentioning

confidence: 99%

Modifications Influencing Widal Test Reactivity in a Novel Microplate Assay

Almogren¹,

Shakoor²,

Adam³

et al. 2012

Pol J Microbiol

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Reliability of the Widal tube agglutination test has been the subject of many controversies over the years. This study was performed to assess the effect of certain modifications on the performance of Widal test in a novel microplate assay. Sera from 37 patients (21 males; 16 females) (mean age 28 +/- 7 years) were tested in the Immunology Unit at King Khalid University Hospital, Riyadh. Among them were 26 patients with suspected typhoid fever and 11 had bacteriologically confirmed diagnosis of Salmonella infection. The modifications included either the use of 0.5% bovine serum albumin (BSA), absorption of sera with sheep red blood cells (SRBC) or heat inactivation of sera. Compared with Widal tube agglutination test, microplate assay with SRBC absorption of the sera from patients with suspected typhoid fever was not only associated with enhancement of detection titers for both H (p < or = 0.001) and O (p < or = 0.005) Salmonella agglutinins but also the percentage of reactivity. The presence of BSA augmented detection titers for Salmonella H agglutinins (p < or = 0.02) only. Heat inactivation of sera however was found to be associated with reduction in the detectable titers for both H (p < or = 0.03) and O (p < or = 0.01) agglutinins. Increased titers of Salmonella agglutinins were also evident in 11 patients with confirmed diagnosis of Salmonella infection. The novel microplate agglutination assay using the SRBC absorption was associated with enhancement in Widal test reactivity and appears to be a useful alternative for the diagnosis of Salmonella infection.

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Serodiagnosis of typhoid fever by enzyme-linked immunosorbent assay determination of anti-Salmonella typhi lipopolysaccharide antibodies

Cited by 43 publications

References 4 publications

Antibodies to porin antigens of Salmonella typhi induced during typhoid infection in humans

Antibodies to porin antigens of Salmonella typhi induced during typhoid infection in humans

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

Modifications Influencing Widal Test Reactivity in a Novel Microplate Assay

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