RETRACTED ARTICLE: Production of recombinant Entamoeba histolyticapyruvate phosphate dikinase and its application in a lateral flow dipstick test for amoebic liver abscess

Saidin, Syazwan; Yunus, Muhammad Hafiznur; Zakaria, Norhayati; Razak, Khairunisak Abdul; Lim, Boon Huat; Othman, Nurulhasanah; Noordin, Rahmah

doi:10.1186/1471-2334-14-182

Cited by 22 publications

(13 citation statements)

References 24 publications

(25 reference statements)

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“…The E. histolytica Gal/Gal-NAc lectin antigen is being utilized in commercial antigen detection tests such as the TechLab E. histolytica II ELISA (TechLab Inc). A study on ES proteins showed the diagnostic potential of pyruvate phosphate dikinase (PPDK), and its recombinant form has been used to develop a lateral flow dipstick test [15, 16]. In terms of treatment, auronofin has been identified as an effective drug which targets thioredoxin reductase, an ES protein of E. histolytica [17].…”

Section: Introductionmentioning

confidence: 99%

Proteome analysis of excretory-secretory proteins of Entamoeba histolytica HM1:IMSS via LC–ESI–MS/MS and LC–MALDI–TOF/TOF

et al. 2016

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BackgroundExcretory-secretory (ES) proteins of E. histolytica are thought to play important roles in the host invasion, metabolism, and defence. Elucidation of the types and functions of E. histolytica ES proteins can further our understanding of the disease pathogenesis. Thus, the aim of this study is to use proteomics approach to better understand the complex ES proteins of the protozoa.Methods E. histolytica ES proteins were prepared by culturing the trophozoites in protein-free medium. The ES proteins were identified using two mass spectrometry tools, namely, LC–ESI–MS/MS and LC–MALDI–TOF/TOF. The identified proteins were then classified according to their biological processes, molecular functions, and cellular components using the Panther classification system (PantherDB).ResultsA complementary list of 219 proteins was identified; this comprised 201 proteins detected by LC–ESI–MS/MS and 107 proteins by LC–MALDI–TOF/TOF. Of the 219 proteins, 89 were identified by both mass-spectrometry systems, while 112 and 18 proteins were detected exclusively by LC–ESI–MS/MS and LC–MALDI–TOF/TOF respectively. Biological protein functional analysis using PantherDB showed that 27% of the proteins were involved in metabolic processes. Using molecular functional and cellular component analyses, 35% of the proteins were found to be involved in catalytic activity, and 21% were associated with the cell parts.ConclusionThis study showed that complementary use of LC–ESI–MS/MS and LC–MALDI–TOF/TOF has improved the identification of ES proteins. The results have increased our understanding of the types of proteins excreted/secreted by the amoeba and provided further evidence of the involvement of ES proteins in intestinal colonisation and evasion of the host immune system, as well as in encystation and excystation of the parasite.Electronic supplementary materialThe online version of this article (doi:10.1186/s12014-016-9135-8) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 99%

Proteome analysis of excretory-secretory proteins of Entamoeba histolytica HM1:IMSS via LC–ESI–MS/MS and LC–MALDI–TOF/TOF

et al. 2016

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“…Thus similar observation may be expected ESA. Western blot analysis using patients serum samples revealed the high diagnostic value of native and recombinant forms of PPDK for detection of ALA [16,17]. Since PPDK is a component of E. histolytica ESA, it is conceivable that it may be also useful as a target for detection of this parasite in stool sample.…”

Section: Discussionmentioning

confidence: 99%

“…Recombinant PPDK (rPPDK) protein was expressed and purified according to our previous report [16]. Meanwhile E. histolytica excretory-secretory antigens (EhESA) was produced using the method we have described earlier [17].…”

Section: Production and Purification Of Polyclonal Antibodiesmentioning

confidence: 99%

Development and initial evaluation of a lateral flow dipstick test for antigen detection ofEntamoeba histolyticain stool sample

Saidin

Yunus

Othman

et al. 2017

Pathogens and Global Health

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Entamoeba histolytica infection remains a public health concern in developing countries. Early diagnosis of amoebiasis can avoid disease complications, thus this study was aimed at developing a test that can rapidly detect the parasite antigens in stool samples. Rabbits were individually immunized with recombinant pyruvate phosphate dikinase (rPPDK) and E. histolytica excretorysecretory antigens to produce polyclonal antibodies. A rapid dipstick test was produced using anti-rPPDK PAb lined on the dipstick as capture reagent and anti-EhESA PAb conjugated to colloidal gold as the detector reagent. Using E. histolytica-spiked in stool sample of a healthy individual, the detection limit of the dipstick test was found to be 1000 cells ml −1. Meanwhile when rPPDK was spiked in the stool sample, the minimum concentration detected by the dipstick test was 0.1 μg ml −1. The performances of the dipstick, commercial Techlab E. histolytica II enzyme-linked immunosorbent assays (ELISA) and real-time PCR were compared using 70 stool samples from patients infected with Entamoeba species (n = 45) and other intestinal pathogens (n = 25). When compared to real-time PCR, the diagnostic sensitivity of the dipstick for detection of E. histolytica was 65.4% (n = 17/26); while the diagnostic specificity when tested with stool samples containing other intestinal pathogens was 92% (23/25). In contrast, Techlab E. histolytica II ELISA detected 19.2% (5/26) of the E. histolytica-positive samples as compared to real-time PCR. The lateral flow dipstick test produced in this study enabled rapid detection of E. histolytica, thus it showed good potential to be further developed into a diagnostic tool for intestinal amoebiasis.

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“…This procedure was similar to our previous report on a dipstick test for the detection of extraintestinal amoebiasis. 25 AgB, as the test line, was jetted linearly onto a Hi-Flow Plus 90 Membrane Card (Millipore, Billerica, MA) using an IsoFlow Dispenser (Imagene Technology, Hanover, NH). Various AgB concentrations (1, 1.2, 1.5, and 2 mg/mL) were dispensed onto separate membrane cards.…”

Section: Methodsmentioning

confidence: 99%

Lateral Flow Test Using Echinococcus granulosus Native Antigen B and Comparison of IgG and IgG4 Dipsticks for Detection of Human Cystic Echinococcosis

Khalilpour¹,

Sadjjadi²,

Moghadam³

et al. 2014

The American Society of Tropical Medicine and Hygiene

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Abstract. Cystic echinococcosis (CE) caused by infection with Echinococcus granulosus is of major concern for humans in many parts of the world. Antigen B was prepared from E. granulosus hydatid fluid, and Western blots confirmed eight batches showing a band corresponding to the 8-/12-kDa subunit with positive serum and no lowmolecular mass band ( 15 kDa) with negative serum. The batches were pooled and used to prepare lateral flow immunoglobulin G4 (IgG4) and IgG dipsticks. Diagnostic sensitivity was determined using serum samples from 21 hydatidosis patients, and diagnostic specificity was established using sera from 17 individuals infected with other parasites and 15 healthy people. IgG4 dipstick had a diagnostic sensitivity of 95% (20 of 21) and a specificity of 100% (32 of 32). The IgG dipstick had a sensitivity of 100% (21 of 21) and a specificity of 87.5% (28 of 32). Thus, both IgG and IgG4 dipsticks had high sensitivities, but IgG4 had greater specificity for the diagnosis of human CE.

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RETRACTED ARTICLE: Production of recombinant Entamoeba histolyticapyruvate phosphate dikinase and its application in a lateral flow dipstick test for amoebic liver abscess

Cited by 22 publications

References 24 publications

Proteome analysis of excretory-secretory proteins of Entamoeba histolytica HM1:IMSS via LC–ESI–MS/MS and LC–MALDI–TOF/TOF

Proteome analysis of excretory-secretory proteins of Entamoeba histolytica HM1:IMSS via LC–ESI–MS/MS and LC–MALDI–TOF/TOF

Development and initial evaluation of a lateral flow dipstick test for antigen detection ofEntamoeba histolyticain stool sample

Lateral Flow Test Using Echinococcus granulosus Native Antigen B and Comparison of IgG and IgG4 Dipsticks for Detection of Human Cystic Echinococcosis

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