Evaluation of a novel proximity ligation assay for the sensitive and rapid detection of foot-and-mouth disease virus.
A novel proximity ligation assay (PLA) using a pan-serotype reactive monoclonal antibody was developed and evaluated for the detection of foot-and-mouth disease virus (FMDV) in clinical samples collected from field cases of disease. The FMDV-specific PLA was found to be 100 times more sensitive for virus detection than the commonly used antigen capture-ELISA (AgELISA). As few as five TCID50 were detected in individual assays, which was comparable with the analytical sensitivity of real-time RT-PCR. Although this assay was capable of detecting diverse isolates from all seven FMDV serotypes, the diagnostic sensitivity of the PLA assay was lower than real-time RT-PCR mainly due to a failure to detect some SAT 1, SAT 2 and SAT 3 FMDV strains. In conclusion, this new PLA format has high analytical sensitivity for the detection of FMDV in clinical samples and may prove valuable as a rapid and simple tool for use in FMD diagnosis.
Nordengrahn
Ann
A
Svanova Biotech AB, Uppsala Science Park, S-751 83 Uppsala, Sweden. ann.nordengrahn@svanova.com
Gustafsdottir
Sigrun M
SM
Ebert
Katja
K
Reid
Scott M
SM
King
Donald P
DP
Ferris
Nigel P
NP
Brocchi
Emiliana
E
Grazioli
Santina
S
Landegren
Ulf
U
Merza
Malik
M
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000302:isolation & purification*; D007118:Immunoassay / Q000379:methods / Q000662:veterinary*; D012367:RNA, Viral / Q000032:analysis; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D012680:Sensitivity and Specificity; D012703:Serotyping; D013997:Time Factors
D000911:Antibodies, Monoclonal; D012367:RNA, Viral