Development and validation of a simplified serotyping ELISA based on monoclonal antibodies for the diagnosis of foot‐and‐mouth disease virus serotypes O, A, C and Asia 1

Abstract

This study describes the development and validation of a simplified enzyme-linked immunosorbent assay (ELISA) for the detection and discrimination of foot-and-mouth disease virus (FMDV) serotypes O, A, C and Asia 1. The multiplex ELISA was designed using selected, type-specific monoclonal antibodies (MAbs) coated onto ELISA plates as catching antibodies and a unique pan-FMDV MAb (1F10) as detector conjugate. Capture MAbs with the broadest intratypic reactivity were selected for each of the four FMDV serotypes by screening large panels of candidate MAbs with a wide spectrum of representative FMDV isolates. An additional pan-FMDV ELISA using 1F10 How to cite this article: Grazioli S, Ferris NP, Dho G, et al. Development and validation of a simplified serotyping ELISA based on monoclonal antibodies for the diagnosis of foot-andmouth disease virus serotypes O, A, C and Asia 1.

Recombinant cystatin-like protein-based competition ELISA for Trichinella spiralis antibody test in multihost sera

“…In Fig 5C , each serum sample of trichinellosis patient was provided two values, one recorded with ES-iELISA and the other one recorded with rCLP-cELISA. The correlation between the two ELISAs was intense (R 2 = 0.7887) [ 29 ], suggesting a direct relationship between OD scores in ES-iELISA and PI ratios in rCLP-cELISA in human sera. A three-band pattern at 45–65 kDa [ 16 , 30 , 32 ] was determined in the ES-WB test using positive sera ( Fig 5D lanes 2–6, black box).…”

Combining Multiple Assays Improves Detection and Serotyping of Foot-and-Mouth Disease Virus. A Practical Example with Field Samples from East Africa

“…The viral RNA, eluted in 60 µL of specific buffer, was stored at −70 • C until used in the experiments. Detection and serotyping of FMDV by Ag-ELISA was carried out using an updated version of the FMDV Antigen Detection and Serotyping ELISA Kit (IZSLER, Brescia, Italy and TPI, Pirbright, UK) [13], including detection and typing of serotypes SAT1 and SAT2, in addition to O, A, C, and Asia 1. Manufacturers' instructions were followed.…”

“…The Ag-ELISA kit incorporating type-specific MAbs, together with a pan-FMDV antigen reaction [13], is the most user-friendly method for serotyping FMD viruses. It can be performed in less than three hours in laboratories with limited infrastructure.…”

“…Recent years have seen many diagnostic advances towards faster detection and serotyping, in order to compensate for some weaknesses of traditional methods [11], including virus isolation (VI) [11,12], Ag-detection ELISA (Ag-ELISA) [11,13], and panreactive real-time RT-PCRs [14][15][16][17]. For example, VI is time consuming, can detect only viable virus, and requires an accompanying test for the identification of the virus; Ag-ELISA has low sensitivity and might fail to detect virus on poor field samples; and no serotyping is possible using pan real-time RT-PCR.…”

“…Type O, A, SAT1, and SAT2 viruses belonging to pool four circulate in this region, and the most recent topotypes affecting this area were A/AFRICA/G-I in 2013, O/EA2 and SAT1/I (NWZ) in 2014, and SAT2/IV in 2012 [25]. The assays included in our comparison were: the real-time RT-PCR targeted on 3D sequence for pan-FMDV detection [14]; a monoclonal antibodies (MAbs)-based FMDV antigen detection and serotyping ELISA (Ag-ELISA) [13]; VI on a continuous porcine cell line [11,26], combined with the above Ag-ELISA; and a panel of four real-time RT-PCRs tailored for the topotypes circulating in the EA region [23]. We report the results of assay comparison, and highlight critical issues emerging during the diagnostic and characterisation process, with particular emphasis on the time required to serotype samples.…”