Diagnostic efficacy of monoclonal antibody based sandwich enzyme linked immunosorbent assay

• Published in: Parasites & vectors 2011-09, Vol.4, p.176
• Main Authors: Demerdash, Zeinab A, Diab, Tarek M, Aly, Ibrahim R, Mohamed, Salwa H, Mahmoud, Faten S, Zoheiry, Mona K, Mansour, Wafaa A, Attia, Mohy E, El-Bassiouny, Azza E
• Format: Article
• Language: English
• Subjects: Care and treatment; Diagnosis; Dosage and administration; Enzyme-linked immunosorbent assay; Fluke infections; Methods; Monoclonal antibodies
• ISSN: 1756-3305; 1756-3305
• DOI: 10.1186/1756-3305-4-176

This research was carried out to develop a reliable monoclonal antibody (MoAb)-based sandwich enzyme linked immunosorbent assay (ELISA) for the diagnosis of active Fasciola gigantica infection in both serum and stool for comparative purposes. From a panel of MoAbs raised against F. gigantica excretory/secretory antigens (ES Ags), a pair (12B/11D/3F and 10A/9D/10G) was chosen due to its high reactivity and strict specificity to F. gigantica antigen by indirect ELISA. The two MoAbs were of the IgG.sub.1 and IgG.sub.2a subclasses, respectively. Using SDS-PAGE and EITB, the selected MoAbs recognized 83, 64, 45 and 26 kDa bands of ES Ags. The lower detection limit of ELISA assay was 3 ng/ml. In stool, the sensitivity, specificity and diagnostic efficacy of ELISA was 96%, 98.2 and 97.1%; while in serum they were 94%, 94.6% and 94.3%, respectively. Moreover, a positive correlation was found between ova count in stool of F. gigantica infected patients and the OD readings of ELISA in both stool and serum samples (r = 0.730, p [less than] 0.01 and r = 0.608; p [less than] 0.01, respectively). These data showed that the use of MoAb-based sandwich ELISA for the detection of F. gigantica coproantigens in stool specimens was superior to serum samples; it provides a highly efficient, non-invasive technique for the diagnosis of active F. gigantica infection.