Trichinella spiralis-specific monoclonal antibodies and affinity-purified antigen-based diagnosis.
Hybridomas secreting monoclonal antibodies (MAbs) to Trichinella spiralis were produced. Myeloma cells were fused with splenocytes of a mouse immunized with excretory-secretory (E-S) antigen of infective larvae. A large percentage of growing hybrids secreted antibodies cross-reactive to many of 23 heterologous parasites tested. Only 6 monoclones (designated 3F2, 5D1, 10F6, 11E4, 13D6 and 14D11) secreted MAbs specific to the E-S antigen and/or a crude extract (CE) of T. spiralis infective larvae. The 6 monoclones secreted IgM, IgG3, IgM, IgG3, IgG3 and IgG3, respectively. Clone 5D1 was selected to mass produce MAbs which were then coupled to CNBr-activated Sepharose CL-4B to prepare an affinity-purified antigen. Dot-blot ELISA with either purified antigen or CE was evaluated. There were 17 patients with acute trichinellosis and 76 individuals convalescing from T. spiralis infection (group 1). Controls were 170 patients with parasitic infections other than trichinellosis (group 2) and 35 healthy parasite-free controls (group 3). CE-ELISA was positive in all group 1 patients. However, sera from many group 2 patients also were reactive (opisthorchiasis-44.2%, schistosomiasis-44%, gnathostomiasis-30%, paragonimiasis-28.6%, taeniasis-27.3%, strongyloidiasis-23.1% and hookworm infections-20%). Affinity-purified antigen was 100% specific, all sera from group 2 and group 3 individuals tested negative. Although 74 of 76 patients (97.4%) with convalescing trichinellosis tested positive, sera from only 3 of 17 patients (17.6%) with acute T. spiralis were reactive. Thus, CE antigen is appropriate when sensitivity is needed, while purified antigen should be used when specificity is required. Dot-blot ELISA is easier to perform, more rapid and less expensive than indirect ELISA. Many samples can be assayed simultaneously, special equipment is not required, and results can be preserved for retrospective analysis. Dot-blot ELISA is therefore the method of choice for the rapid diagnosis of trichinellosis, particularly when more complex laboratory tests are unavailable.
Srimanote
P
P
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, 10400 Thailand.
Ittiprasert
W
W
Sermsart
B
B
Chaisri
U
U
Mahannop
P
P
Sakolvaree
Y
Y
Tapchaisri
P
P
Maleewong
W
W
Kurazono
H
H
Hayashi
H
H
Chaicumpa
W
W
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000302:isolation & purification; D016022:Case-Control Studies; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000706:statistics & numerical data; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007159:Immunologic Tests; D051379:Mice; D012680:Sensitivity and Specificity; D017160:Trichinella spiralis / Q000276:immunology*; D014235:Trichinellosis / Q000175:diagnosis* / Q000276:immunology
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth