Generation of monoclonal antibodies against foot‐and‐mouth disease virus SAT 2 and the development of a lateral flow strip test for virus detection

Abstract

Foot‐and‐mouth disease (FMD) remains a major economic concern for the livestock productivity in many developing countries and a continued threat to countries that are disease free because of its potential devastating impact on agricultural, food chain and tourism sectors. FMD virus (FMDV) is recognized as having seven serotypes: O, A, C, Asia 1, South African Territories (SAT) 1, 2, 3 and multiple subtypes within each serotype. FMD outbreaks due to SAT 2 have been reported in many African countries. The development of a rapid and easily performed test for FMD detection is critical for controlling FMD outbreaks and containing its spread. The present project developed a lateral flow immunochromatographic (LFI) strip test for the rapid detection of FMDV SAT 2. A panel of monoclonal antibodies (mAbs) against FMDV serotype SAT 2 was produced and characterized. One mAb (#10) was selected as the capture mAb because it reacted to all 23 SAT 2 isolates archived at the National Center for Foreign Animal Disease. The LFI strip test was developed using biotin‐conjugated mAb #10, and the colloid gold‐conjugated FMDV serotype‐independent mAb as the detection mAb. A generic Rapid Assay Device (gRAD) with one test line and a control line was used for the test. The LFI strip test detected all 23 tested SAT 2 isolates and recent outbreak strains. The results indicated that the diagnostic specificity and sensitivity of the LFI strip test were greater than the double antibody sandwich (DAS) DAS ELISA. The ability of the LFI strip test to produce rapid diagnostic results will be useful for early on‐site diagnosis during FMD outbreaks.

Detection of Foot-and-Mouth Disease Virus in Swine Meat Juice

“…However, antigen detection by LFI was short-lived compared to rRT-PCR, implying that LFI might fail to detect low levels of viral antigen in MJ. rRT-PCR is inherently a more sensitive assay [ 20 ] and thus explains the detection of FMDV RNA over a wider duration of the experiment. The fact that LFI can detect FMDV in MJ is a good reason to collect both MJ and sera when possible, considering that FMDV antigen detection in serum samples by LFIs has not been successful [ 21 ].…”

“…Meat juice samples from naïve and FMDV–infected pigs were tested for antigen by LFI for FMDV serotype A [ 21 ] and FMDV serotype SAT 2 [ 20 ] following the published protocols for each serotype. Each test utilizes two conjugated serotype-specific monoclonal antibodies.…”

“…One antibody is conjugated to biotin, which is captured at the test line by streptavidin, while the other antibody is conjugated to gold for visualization of bound antigen. Limit of detection for the test is 10 3.77 TCID 50 for serotype SAT2 and 10 5.50 for serotype A [ 20 , 21 ].…”

Phage Display Screening of Bovine Antibodies to Foot-and-Mouth Disease Virus and Their Application in a Competitive ELISA for Serodiagnosis

“…Diverse sets of MAbs against FMDV have been generated by murine hybridoma technology [ 7 , 8 , 9 , 10 ]. Extensive studies on antigenic features of FMDV, especially type O, have identified major neutralizing epitopes recognized by the mouse MAbs [ 11 , 12 , 13 ].…”

Foot‐and‐mouth disease virus detection on a handheld real‐time polymerase chain reaction platform

“…Antigen detection and molecular assays have been evaluated for this purpose. Lateral flow strip tests (LFSTs) for FMDV antigen detection are available, some of which are being used in the field (Ferris et al, , ; Yang, Caterer, Xu, & Goolia, ; Yang, Goolia, Xu, Bittner, & Clavijo, ; Yang, Mudabuka, Quizon, & Nfon, ). LFST was used for FMD detection during the 2007 FMD outbreak in the UK (Ryan et al, ).…”