title: Diagnostic efficacy of monoclonal antibody based sandwich enzyme linked immunosorbent assay (ELISA) for detection of Fasciola gigantica excretory/secretory antigens in both serum and stool.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  The two MoAbs were of the IgG₁ and IgG(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 < 0.01 and r = 0.608; p < 0.01, respectively).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Immunology, Theodor Bilharz Research Institute, Giza, Egypt.
authors.forename: Zeinab A
authors.initials: ZA
authors.lastname: Demerdash
authors.forename: Tarek M
authors.initials: TM
authors.lastname: Diab
authors.forename: Ibrahim R
authors.initials: IR
authors.lastname: Aly
authors.forename: Salwa H
authors.initials: SH
authors.lastname: Mohamed
authors.forename: Faten S
authors.initials: FS
authors.lastname: Mahmoud
authors.forename: Mona K
authors.initials: MK
authors.lastname: Zoheiry
authors.forename: Wafaa A
authors.initials: WA
authors.lastname: Mansour
authors.forename: Mohy E
authors.initials: ME
authors.lastname: Attia
authors.forename: Azza E
authors.initials: AE
authors.lastname: El-Bassiouny
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000947:Antigens, Helminth / Q000032:analysis* / Q000097:blood*; D019411:Clinical Laboratory Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005211:Fascioliasis / Q000175:diagnosis*; D005243:Feces / Q000737:chemistry; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity; D044967:Serum / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D007074:Immunoglobulin G