Development and initial evaluation of a lateral flow dipstick test for antigen detection of Entamoeba histolytica in stool sample. (3rd April 2017)

• Record Type: Journal Article
• Authors: Saidin, Syazwan; Yunus, Muhammad Hafiznur; Othman, Nurulhasanah; Lim, Yvonne Ai-Lian; Mohamed, Zeehaida; Zakaria, Nik Zairi; Noordin, Rahmah
• Is Part Of: Pathogens and global health. Volume 111:Number 3(2017)
• Page Start: 128
• Page End: 136
• Publication Date: 2017-04-03
• Subjects: Entamoeba histolytica -- stool antigen detection -- pyruvate phosphate dikinase -- excretory-secretory antigens -- lateral flow rapid test; Communicable diseases -- Periodicals; Public health -- International cooperation -- Periodicals; World health -- Periodicals; 362.1969
• DOI: 10.1080/20477724.2017.1300421
• Languages: English
• ISSNs: 2047-7724

Abstract

Abstract: 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 excretory-secretory 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.