Serodiagnosis of Extraintestinal Amebiasis: Retrospective Evaluation of the Diagnostic Performance of the Bordier® ELISA Kit

Abstract

Soluble antigens from an axenic culture of Entamoeba histolytica were used to develop a commercial ELISA kit to quantify anti-E. histolytica antibodies in sera of patients with extraintestinal amebiasis in non-endemic settings. The diagnostic specificity and sensitivity of the test were assessed retrospectively using 131 human serum samples with amoebic serologic status available. They were selected according to their results in immunofluorescence (IFAT) and were separated in 2 sample categories: 64 sera with positive results by IFAT and 67 with negative results by IFAT. The sensitivity and specificity of the ELISA kit were assessed at 95.0% and 94.0% compared to the IFAT. The test can be useful to exclude a potential diagnosis of amebiasis and could be used as a screening method since ELISA is an automated technique.

Evaluation of total immunoglobulin G and subclass antibodies in an enzyme-linked immunosorbent assay for serodiagnosis of human amebic liver abscess

“…The present study was designed to assess the diagnostic potential of ELISA, targeting total IgG and IgG subclasses produced against crude soluble antigens of E. histolytica . Our IgG-based ELISA showed a high diagnostic sensitivity (100%) and specificity (97.8%), comparable to the values of 80–100% sensitivities and 93–100% specificities reported for commercial and in-house IgG ELISA kits in cases of ALA ( Tanyuksel & Petri Jr, 2003 ; Knappik, Borner & Jelinek , 2005 ; Wong et al, 2017 ; Beyls et al, 2018 ; Saidin, Othman & Noordin, 2019 ). Interestingly, group 3 contains sera from patients with significant parasitic diseases prevalent in the developing world, implying that these patients are hugely affected by protozoa, including E. histolytica , on a regular basis.…”

“…Polymerase chain reaction (PCR) assay, the gold standard method, helps diagnose ALA if aspirated pus is available, but this technique is costly and only used in some laboratories, particularly in developed countries, where infrastructure and skillful staff are available ( Zaman et al, 2000 ; Tanyuksel & Petri Jr, 2003 ; Ryan, Paparini & Oskam, 2017 ; Saidin, Othman & Noordin, 2019 ). Therefore, in practice, the diagnosis of ALA is based on clinical features, history of living in or traveling to endemic regions, imaging techniques such as ultrasound (US), computerized tomography (CT), and magnetic resonance imaging (MRI), as well as serological tests ( Wong et al, 2017 ), like immunofluorescence (IF) tests ( Garcia et al, 1982 ; Jackson, Anderson & Simjee, 1984 ), indirect hemagglutination assays (IHA) ( Knobloch & Mannweiler, 1983 ; Hung et al, 1999 ; Dhanalakshmi, Meenachi & Parija, 2016 ), enzyme immunoassays ( Yang & Kennedy, 1979 ; Hock et al, 1989 ; Shetty et al, 1990 ; Knappik, Borner & Jelinek , 2005 ; Wong et al, 2017 ; Beyls et al, 2018 ), and lateral flow immunoassays ( Saidin et al, 2014 ; Tachibana et al, 2018 ; Noordin et al, 2020 ).…”

“…For ELISA, most of the circulating E. histolytica -specific antibody detection tests indicate using immunoglobulin G (IgG) in the detection system ( Yang & Kennedy, 1979 ; Knappik, Borner & Jelinek, 2005 ; Fotedar et al, 2007 ; Wong et al, 2017 ; Beyls et al, 2018 ; Saidin, Othman & Noordin, 2019 ; Watanabe et al, 2021 ) because invasive amebic infections provoke a strong humoral response, especially involving the IgG class ( Hock et al, 1989 ; Shetty et al, 1990 ). Anti-amebic IgG and its subclasses were determined by ELISA in the sera of ALA patients and compared with those from normal healthy controls.…”

Multi-Laboratory Evaluation of a Lateral Flow Rapid Test for Detection of Amebic Liver Abscess

“…27 Entamoeba histolytica IgG ELISA (Bordier) performed on biobank serum samples at a laboratory in Grenoble, France, showed a sensitivity of 95% (61/64) and a specificity of 94% (63/67) as compared with previously available indirect fluorescent antibody test (IFAT) results. 28 A prototype lateral flow test, XEh Rapid, an IgG4-based assay using PPDK recombinant protein, has been developed for the detection of ALA. 19 Here, we present the results of an independent evaluation of the performance of XEh Rapid in seven laboratories in four countries, that is, Malaysia, Iran, Australia, and Spain. The test showed high diagnostic sensitivity of 89-91% at three of the five laboratories, thus almost comparable to the diagnostic sensitivity reports of commercial IgG-ELISA kits described earlier, with the additional benefit of being a lateral flow rapid test.…”

Immunoblot for the Diagnosis of Cutaneous Leishmaniasis in French Guiana

“…However, the Leishmania IB cross-reaction with extraintestinal amoebiasis was unexpected and, to the best of our knowledge, has never been reported. 20 , 21 This disease has a distinct clinical presentation and is not a differential diagnosis of CL; this cross-reactivity was unexpected because, although both are protozoa, Entamoeba and Leishmania are phylogenetically distant genera, whereas Trypanosoma and Leishmania belong to the same Euglenozoa phylum. To verify this cross-reaction, Leishmania IB was carried out on eight sera of patients with proven extraintestinal amoebiasis from Marseille (laboratory of Parasitology Mycology), a non-endemic region for New-World CL, and none of the tested sera will have expressed p14 and/or p16 antigens.…”