Detection of Giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies

Duque-Beltrán, Sofía; Nicholls-Orejuela, Rubén Santiago; Arévalo-Jamaica, Adriana; Guerrero‐Lozano, Rafael; Montenegro, Sonia; James, Mark A.

doi:10.1590/s0074-02762002000800018

Cited by 19 publications

(12 citation statements)

References 20 publications

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“…A combination of the PGA with complete and incomplete Freund's adjuvant was made to immunize rabbits and prepare rabbit anti-G. lamblia IgG pAb 32 . Many purposes were detected form usage of adjuvants in animal's immunization protocols including enhancement of the immune response to the antigen, furthermore introducing a depot for the immunogens at the injection site allowing for slow, prolonged release of the immunogens in the animal, this agreed a previously reported work 33 .…”

Section: Discussionsupporting

confidence: 81%

Combined Mini-Parasep SF and Nanogold Immunoassay Show Potential in Stool Antigen Immunodetection for Giardiasis Diagnosis

Mohram

Elawamy

Nageeb

et al. 2020

Sci Rep

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covalent loading or directional binding of biomolecules on gold nanoparticles (Aunps) could lead to better results than simple direct adsorption for an enhanced eLiSA application. the use of Mini-parasep solvent-free (SF) without ether or ethyl acetate for the clean and efficient concentration of protozoa cysts, it is a single-use device for in vitro diagnostic use only. in this work, we used Mini-parasep Sf for the detection of giardia cysts in comparison to direct smear and Merthiolate-iodine formaldehyde concentration (Mifc) technique in addition to its use in antigen detection by Aunps biomolecule loading using rabbit polyclonal antibodies (pAb) against purified Giardia antigen (pGA). As a result, Mini-Parasep SF was the most effective method for Giardia cyst detection and regarding optimization of Mini-Parasep antigen detection, our data showed increased sensitivity and specificity of nano-sandwich ELISA to 92% and 94% respectively and increased positive predictive value (PPV) and negative predictive value (NPV) to 88.64% and 95.91% respectively. In conclusion, this research provides that Mini-parasep Sf concentrator enhanced Giardia cyst detection and improved antigen preparation for Aunps sandwich eLiSA in giardiasis diagnosis. the advantages of this method are the short assay time and the raised accuracy of antigen detection providing concentrated samples without the risk of solvent use and being a disposable Mini-parasep it helps in giardia antigen purification as well as raising the sensitivity and specificity of ELISA through binding AuNPs.

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Section: Discussionsupporting

confidence: 81%

Combined Mini-Parasep SF and Nanogold Immunoassay Show Potential in Stool Antigen Immunodetection for Giardiasis Diagnosis

Mohram

Elawamy

Nageeb

et al. 2020

Sci Rep

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“…The final suspension in this method had a little amount of extras. In another study that was at 2002 to produce polyclonal antibodies against Giardia antigens, the mentioned purification method were used (Duque-Beltran et al, 2002). Percoll solution is very expensive and because of its uncommon usage, is commercially scarce.…”

Section: Discussionmentioning

confidence: 99%

“…Routine method to diagnose giardiasis is direct microscopic examination of stool (Rosoff and Stibbs, 1986;Duque-Beltran et al, 2002). Sensitivity of microscopic method even with several times repeating of each test is just 50 to 70% (Burke et al, 1975(Burke et al, , 1977.…”

Section: Introductionmentioning

confidence: 99%

Introducing a simple and economical method to purify Giardia lamblia cysts

Barazesh¹,

Majidi²,

Fallah³

et al. 2011

Afr. J. Biotechnol.

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Direct microscopic examination of stool to diagnosis giardiasis (wet mount) has low diagnostic value, but immunologic methods (like IFA and especially ELISA) that are based on the determination of parasite antigens in fecal samples (antigen detection) have relatively high sensitivity and specialty. To prepare anti-Giardia lamblia antibodies needed to design diagnostic kits as well as parasite culture and other molecular studies, we require purification of the parasite cysts. In this study, we designed a rapid, simple and inexpensive method to purify parasite cysts from fecal samples of the patients suffering from giardiasis. Initially, fecal samples that the presence of G. lamblia in them was affirmed by direct microscopic observation of cysts were subjected to various purification methods like one-and twophase sucrose gradient isolation, percoll-sucrose gradient isolation, and a modified two-phase method run by 0.85 and 1.5 M sucrose. The first procedure contained some contents of bacteria and small particles of feces. In the second and third procedure, bacteria were almost removed and the cysts were intact but the suspension contained some extras and cellulose particles. The recovery rate for modified two-phase method was 1.5 × 10 4 cysts for each two grams of fecal sample. In this study, by using and comparing with the results of some other studies, we introduce and run a modified method that in fact is a mélange of them with some changes. So this method could be recommended as a fast, advantageous and simple method in purification of G. lamblia cysts.

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“…The diagnosis of Giardiasis has been classically on detection of cysts or trophozoites in stool and duodenal aspirates specimens by direct microscopic examination and duodenal endoscopic biopsy specimen by histologically. Recently proven, ELISA methods for the detection of Giardia-specific antigen in fecal specimens have 97.5% sensitivity and 99.5% specificity [14, 15]. …”

Section: Introductionmentioning

confidence: 99%

The Prevalence of Giardia Intestinalis in Dyspeptic and Diabetic Patients

Hakim

Kızıltaş

Çiftçi

et al. 2011

ISRN Gastroenterology

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Background and Aims. We aimed to investigate the prevalence of Giardiasis in patients with dyspepsia and patients with diabetes mellitus. Methods. 400 patients and 100 healthy persons were included in this clinical prospective study. The number of patients in each group was equal, 200 dyspeptic and 200 diabetic, respectively. The antigen of G. lntestinalis was determined in the stool specimens by ELISA method. Results. The frequency of Giardiasis was 7% in dyspeptic and 15% in diabetic patients. There was no positive results in any of the healthy persons. There was a significant difference in prevalence rate of Giardiasis between patients with dyspepsia and diabetes mellitus (P < 0.05). Conclusions. These results revealed that the prevalence of Giardiasis in dyspepsia and with diabetes mellitus was high in our country. This is the first study investigating the prevalence of Giardiasis in diabetic patients. To investigate Giardiasis in diabetic patients, who have dyspepsia or not, may be a good approach for public health.

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Detection of Giardia duodenalis antigen in human fecal eluates by enzyme-linked immunosorbent assay using polyclonal antibodies

Abstract: The present study developed and standardized an enzime-linked immunosorbent assay (ELISA)

Cited by 19 publications

References 20 publications

Combined Mini-Parasep SF and Nanogold Immunoassay Show Potential in Stool Antigen Immunodetection for Giardiasis Diagnosis

Combined Mini-Parasep SF and Nanogold Immunoassay Show Potential in Stool Antigen Immunodetection for Giardiasis Diagnosis

Introducing a simple and economical method to purify Giardia lamblia cysts

The Prevalence of Giardia Intestinalis in Dyspeptic and Diabetic Patients

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