Cystatin capture-dot-enzyme-linked immunosorbent assay for immunodiagnosis and assessment of cure of experimental trichinellosis in mice.
Cystatin capture dot enzyme-linked immunosorbent assay (CC-dot ELISA) was evaluated as a new version of ELISA for diagnosis of trichinellosis by the detection of anti-Trichinella spiralis cysteine proteinase (CyP) IgG, using nitrocellulose membrane sensitized with cystatin as a capture reagent for CyP from T. spiralis muscle larvae excretory secretory products (ESP) without purification, compared to the detection of anti-T. spiralis IgG by conventional (conv.) ELISA, using whole ESP. Experimentally infected mice with light (100 larvae/mouse) and heavy (300 larvae/mouse) infections by T. spiralis larvae at 7, 14, 21 and 56 days post infection, and after flubendazole treatment were examined. As early as one week post infection CC-dot ELISA gave high positive rates of 86.6% and 100% in light and heavy infections, respectively, in contrast to negative results by conv. ELISA. CC-dot ELISA showed in light and heavy trichinellosis a higher efficiency in comparison to conv. ELISA (97.7% versus 67.7% and 98.8% versus 80%, respectively) and a higher overall sensitivity (96.6% versus 55% and 98.3% versus 73.3%, respectively). No cross reactions with sera of other parasitic infected or non infected control mice were recorded by CC-dot ELISA giving 100% specificity compared to 93.3% by conv. ELISA. After treatment, CC-dot ELISA gave positive results only in uncured mice with remaining muscle larvae, while conv. ELISA was positive in mice with and without remaining muscle larvae. CC-dot ELISA used lower quantities of antigen, was performed at room temperature, and read by naked eye in less than 2.5 hours.
Department of Parasitology, Faculty of Medicine, Ain Shams University, Cairo 11566, Egypt.
Manal S E
MS
Mahmoud
Manal A
MA
Moustafa
D000818:Animals; D000969:Antinematodal Agents / Q000627:therapeutic use; D015891:Cystatins / Q000276:immunology*; D003546:Cysteine Endopeptidases / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D007074:Immunoglobulin G / Q000032:analysis / Q000097:blood; D008463:Mebendazole / Q000031:analogs & derivatives* / Q000627:therapeutic use; D051379:Mice; D012680:Sensitivity and Specificity; D017160:Trichinella spiralis / Q000187:drug effects; D014235:Trichinellosis / Q000175:diagnosis* / Q000188:drug therapy* / Q000276:immunology
D000969:Antinematodal Agents; D015891:Cystatins; D007074:Immunoglobulin G; D008463:Mebendazole; D003546:Cysteine Endopeptidases; C018945:flubendazole