Development and laboratory evaluation of a lateral flow device (LFD) for the serodiagnosis of Theileria annulata infection.
Several DNA-based and serological tests have been established for the detection of Theileria annulata infection, including polymerase chain reaction, reverse line blot and loop-mediated isothermal amplification, indirect enzyme-linked immunosorbent assay (ELISA), and competitive ELISA. In this study, we have applied knowledge from the development and application of a recombinant protein-based indirect ELISA and competitive ELISA to establish a rapid test for point-of-care diagnosis of T. annulata infection in the field to be used by the veterinarian. For the development of a lateral flow test, the recombinantly expressed T. annulata surface protein (TaSP) was applied as the test antigen and anti-TaSP antiserum as the control line. TaSP antigen conjugated to colloidal gold particles was used as the detection system for visualization at the test line for the binding of anti-TaSP antibody present in the serum of infected animals. The developed test specifically detected antibodies in the serum of animals experimentally infected with T. annulata and showed no cross-reactivity with serum from animals infected with other tested bovine pathogens (Trypanosoma brucei, Anaplasma marginale, Babesia bigemina, Babesia bovis, and Theileria parva). Testing of field samples was compared to results obtained by other serological tests, resulting in a sensitivity and specificity of 96.3% and 87.5% compared to indirect fluorescence antibody test, 98.7% and 81.8% compared to indirect ELISA, and 100% and 47.6% compared to competitive ELISA. In conclusion, a rapid test for the detection of T. annulata infection (T. annulata lateral flow device, Ta-LFD) has been developed, which is easy to perform, delivers results to be read by the naked eye within 10 min, and is suitable for the detection of infection in field samples.
Abdo
Jassim
J
Division of Veterinary Infection Biology and Immunology, Research Center Borstel, Parkallee 22, 23845 Borstel, Germany.
Kristersson
Therese
T
Seitzer
Ulrike
U
Renneker
Stefanie
S
Merza
Malik
M
Ahmed
Jabbar
J
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood*; D000953:Antigens, Protozoan; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology; D010274:Parasitology / Q000379:methods* / Q000592:standards; D019095:Point-of-Care Systems / Q000592:standards; D011994:Recombinant Proteins; D012015:Reference Standards; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods / Q000592:standards; D016796:Theileria annulata / Q000276:immunology* / Q000302:isolation & purification; D013801:Theileriasis / Q000175:diagnosis* / Q000469:parasitology
D000909:Antibodies, Helminth; D000953:Antigens, Protozoan; D011994:Recombinant Proteins