Use of a Novel Enzyme Immunoassay Based on Detection of Circulating Antigen in Serum for Diagnosis of<i>Helicobacter pylori</i>Infection

Attallah, Abdelfattah M.; Ismail, Hisham; Ibrahim, Gellan G.; Abdel-Raouf, Mohamed; El-Waseef, Ahmed; Wahab, Mohamed Abdel

doi:10.1128/cdli.11.4.775-779.2004

Cited by 11 publications

(16 citation statements)

References 34 publications

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“…Rabbit immunization was done as previously described (Attallah et al 2004). In brief, the rabbit was immunized subcutaneously at three different inoculation sites with 500 μg of recombinant GST-TgFABZ protein diluted in an equal volume of Freund's complete adjuvant.…”

Section: Generation Of Polyclonal Anti-tgfabz and Anti Tgacp Antibodiesmentioning

confidence: 99%

Molecular and biochemical characterization of Toxoplasma gondii β-hydroxyacyl-acyl carrier protein dehydratase (FABZ)

et al. 2008

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Toxoplasma gondii, unlike its mammalian host, utilizes a type II fatty acid biosynthesis pathway in which the steps of fatty acid biosynthesis are catalyzed by independent enzymes. Due to this difference, the enzymes of this pathway are good targets for the development of new therapeutic drugs directed against toxoplasmosis. In this report, we show by using reverse transcription-polymerase chain reaction analysis that beta-Hydroxyacyl-acyl carrier protein dehydratase (TgFABZ) is expressed both in tachyzoites and bradyzoites. Indirect immunofluorescence antibody test further shows the localization of TgFABZ protein in the apicoplast of both tachyzoites and bradyzoites. Enzyme dynamic analysis shows that the purified recombinant TgFABZ protein is soluble and active. The Km value of the enzyme for its substrate analog crotonoyl-CoA was estimated to be 82.57 +/- 10 microM.

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Section: Generation Of Polyclonal Anti-tgfabz and Anti Tgacp Antibodiesmentioning

confidence: 99%

Molecular and biochemical characterization of Toxoplasma gondii β-hydroxyacyl-acyl carrier protein dehydratase (FABZ)

et al. 2008

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“…Then normal goat serum diluted (1:5) with 1-4% bovin serum albumin (BSA) in tris-buffered saline (TBS, pH 7.2) was applied and incubated for 60 min. Excess liquid was wiped away and specific IgG rabbit serum to HpLysate or to the 58-kDa H. pylori antigen [17] was then applied an incubated for 1 hr. After washing, horseradish peroxidase conjugated goat antibody to rabbit immunoglobulins (Sigma, CA, USA) diluted 1:500 in 1.5% BSA=TBS and incubated for 60 min.…”

Section: Histopathological Examinationmentioning

confidence: 99%

“…[18] The microorganism was identified as H. pylori using the standard methods on the basis of colony morphology, gram stain, and the production of urease, catalse, and oxidase enzymes [19] and further confirmed by the Western blot using rabbit antiserum against whole cell lysate of H. pylori. [17] The human pathogenic H. pylori strain was adapted in a group of BALB=c female mice (6 week-old and weighing $20 g) for 2 weeks. Mice were given 0.25 mL of 0.2 M NaHCO 3 to neutralize acidity, then the bacterial inoculum was administered as a single dose of 1 Â 10 8 CFU=mL in 0.15 mL of sterile saline by gastric intubation with a blunt needle attached to a syringe under light anesthesia with halothane (Halocarbon, River Edge, NJ, USA).…”

Section: H Pylori Pathogenic Strainmentioning

confidence: 99%

“…[16] Recently, we have identified a highly reactive and specific 58-kDa antigen in whole cell lysate antigenic extract of H. pylori and evaluateed its diagnostic potential. [17] The present work aimed to establish a stable and reliable model of H. pylori infection in BALB=c mice using a human pathogenic strain of H. pylori to produce specific IgY antibodies from egg yolk of hens immunized with H. pylori antigens and to evaluate its efficacy in passive immunization therapy for inhibition of H. pylori infection and gastritis.…”

Section: Introductionmentioning

confidence: 99%

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Efficacy of Passive Immunization with IgY Antibodies to a 58-kDaH. pyloriAntigen on Severe Gastritis in BALB/c Mouse Model

Attallah

Abbas

Ismail

et al. 2009

Journal of Immunoassay and Immunochemistry

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Consecutive triple doses of 1 x 10(8) CFU/mL of a pathogenic H. pylori strain isolated from stomach of Egyptian patients with severe gastritis were used to establish infection in BALB/c mice model. White Leghorn hens were immunized with H. pylori whole cell lysate (HpLysate) antigen and with a highly reactive 58-kDa H. pylori (Hp58) antigen. Two months later, IgY antibodies (IgY-HpLysate & IgY-Hp58) were purified from egg yolk and its efficacy was evaluated in the adopted model. Microbiological culture and immunohistochemical staining revealed that H. pylori infection was inhibited 1 week after oral passive immunization in 70% of infected BALB/c mice with a significant decrease (p < 0.05) in the degrees of gastritis. In conclusion, we have adapted BALB/c mice model for human H. pylori pathogenic strain and oral passive immunization with specific IgY antibodies to the 58-kDa antigen inhibited active H. pylori infection and decreased gastritis.

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“…The human E-cadherin EIA kit is a solid phase EIA based on a sandwich method that utilizes 2 mouse monoclonal anti-human E-cadherin antibodies to detect soluble E-cadherin in a 2-step procedure [15]. [16][17][18].…”

Section: Patients and Samplesmentioning

confidence: 99%

501 Evaluation of specific biochemical indicators of Helicobacter pylori-associated gastric cancer in Egypt

Anwar¹,

Youssef²,

Sheta³

et al. 2012

East Mediterr Health J

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The aim of the study was to assess the accuracy of some specific biochemical indicators in discriminating between Helicobacter pylori-associated gastritis and H. pylori-associated stomach cancer (serum gastrin level, serum soluble E-cadherin and tissue COX-2 activity, as well as serodiagnostic markers for H. pylori infection) in order to find a simple diagnostic test that can reasonably predict the development of gastric cancer. The study participants comprised 20 patients with gastric carcinoma, 20 patients with positive H. pyloriassociated gastritis and 20 individuals as the control group. Standard procedures and quality control measures were followed. Using cut-off values and ROC analysis to assess the diagnostic abilities of the biochemical indicators, E-cadherin showed the highest sensitivity (100%). We suggest that close follow-up together with periodic endoscopic examination for all patients with persistent H. pylori infection and serum soluble E-cadherin level above 5 µg/mL is essential. Après l'utilisation de valeurs seuils et d'une analyse de la fonction d'efficacité du récepteur pour évaluer les qualités diagnostiques des indicateurs biologiques, la E-cadhérine a montré la sensibilité la plus élevée (100 %). Nous suggérons qu'un suivi attentif et un examen endoscopique régulier sont essentiels pour tous les patients atteints d'une infection à H. pylori persistante et présentant une concentration sérique de la E-cadhérine soluble supérieure à 5 µg/ml.

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Use of a Novel Enzyme Immunoassay Based on Detection of Circulating Antigen in Serum for Diagnosis ofHelicobacter pyloriInfection

Cited by 11 publications

References 34 publications

Molecular and biochemical characterization of Toxoplasma gondii β-hydroxyacyl-acyl carrier protein dehydratase (FABZ)

Molecular and biochemical characterization of Toxoplasma gondii β-hydroxyacyl-acyl carrier protein dehydratase (FABZ)

Efficacy of Passive Immunization with IgY Antibodies to a 58-kDaH. pyloriAntigen on Severe Gastritis in BALB/c Mouse Model

501 Evaluation of specific biochemical indicators of Helicobacter pylori-associated gastric cancer in Egypt

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