Co-infection with different serotypes of FMDV in vaccinated cattle in Southern Egypt.

During 2015-2016 period, an outbreak of foot-and-mouth disease virus (FMDV) was observed in cattle in four governorates of the upper of Egypt. The infection was extended to the vaccinated cattle. A total of 54 mouth swabs and serum samples were collected from vaccinated cattle for serological and virological investigation.

Assessment of the First Commercial ELISA Kit for the Diagnosis of Theileria annulata.

The present study assesses the efficacy of SVANOVIR Theileria annulata-Ab, the first commercial ELISA kit for the diagnosis of Theileria annulata infection in cattle based on a recombinant protein known as T. annulata surface protein (TaSp). As a reference test, a polymerase chain reaction (PCR) assay depending on T.

Development and laboratory evaluation of two lateral flow devices for the detection of vesicular stomatitis virus in clinical samples.

Two lateral flow devices (LFD) for the detection of vesicular stomatitis (VS) virus (VSV), types Indiana (VSV-IND) and New Jersey (VSV-NJ) were developed using monoclonal antibodies C1 and F25VSVNJ-45 to the respective VSV serotypes. The performance of the LFDs was evaluated in the laboratory on suspensions of vesicular epithelia and cell culture passage derived supernatants of VSV. The collection of test samples included 105 positive for VSV-IND (92 vesicular epithelial suspensions and 13 cell culture antigens; encompassing 93 samples of subtype 1 [VSV-IND-1], 9 of subtype 2 [VSV-IND-2] and 3 of subtype 3 [VSV-IND-3]) and 189 positive for VSV-NJ (162 vesicular epithelial suspensions and 27 cell culture antigens) from suspected cases of vesicular disease in cattle and horses collected from 11 countries between 1937 and 2008 or else were derived from experimental infection and 777 samples that were either shown to be positive or negative for foot-and-mouth disease (FMD) virus (FMDV) and swine vesicular disease virus (SVDV) or else collected from healthy cattle or pigs and collected from 68 countries between 1965 and 2011.

Evaluation of a blocking ELISA for the detection of antibodies against Lawsonia intracellularis in pig sera.

Background: Lawsonia intracellularis is a common cause of chronic diarrhoea and poor performance in young growing pigs. Diagnosis of this obligate intracellular bacterium is based on the demonstration of the microbe or microbial DNA in tissue specimens or faecal samples, or the demonstration of L. intracellularis-specific antibodies in sera.

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.

A microsphere-based immunoassay for rapid and sensitive detection of bovine viral diarrhoea virus antibodies.

This study describes a novel blocking microsphere-based immunoassay for highly sensitive and specific detection of antibodies against bovine viral diarrhoea virus (BVDV). The intra- and inter-assay variability are 4.9% and less than 7%, respectively, and variability of bead conjugations is less than 6.

Development and laboratory evaluation of a lateral flow device for the detection of swine vesicular disease virus in clinical samples.

A lateral flow device (LFD) for the detection of swine vesicular disease (SVD) virus (SVDV) and differential diagnosis from foot-and-mouth disease (FMD) was developed using a monoclonal antibody (Mab C70). The performance of the LFD was evaluated in the laboratory on suspensions of vesicular epithelia and cell culture passage derived supernatants of SVDV and porcine teschovirus (enterovirus; PEV). The collection of test samples included 157 which were positive for SVDV (84 vesicular epithelial suspensions and 73 cell culture antigens) from suspected cases of vesicular disease in pigs collected from 14 countries between 1966 and 2008 and 663 samples which were either shown to be negative for SVDV and FMD virus (FMDV) or else collected from healthy pigs or demonstrated to be positive for FMDV, PEV or vesicular exanthema (VEV) and collected from 16 countries between 1965 and 2008 or else were derived from experimental animals.

Development and laboratory validation of a lateral flow device for the detection of serotype SAT 2 foot-and-mouth disease viruses in clinical samples.

A lateral flow device (LFD) for the detection of foot-and-mouth disease virus (FMDV) of the SAT 2 serotype was developed using a monoclonal antibody (Mab 2H6). The performance of the LFD was evaluated in the laboratory on suspensions of vesicular epithelia: 305 positive for FMDV type SAT 2 from suspected cases of vesicular disease collected from 30 countries and 1002 samples shown to be negative for FMDV type SAT 2 collected from 67 countries between 1968 and 2008. The diagnostic sensitivity of the LFD for FMDV type SAT 2 was higher at 88% compared to 79% obtained by the reference method of antigen ELISA, and the diagnostic specificity of the LFD was approximately 99% compared to 100% for the ELISA.

Assessment of the within- and between-laboratory repeatability of a commercially available Ostertagia ostertagi milk ELISA.

The objectives of this study were (1) to assess the within- and between-laboratory repeatability of a commercially available antibody-detection Ostertagia ostertagi ELISA (SVANOVIR(O). ostertagi-Ab, Svanova, Uppsala) and (2) to investigate if the assay could be further simplified by reading optical density at a single instead of double wavelength. A total of 80 bulk-tank milk samples were divided into aliquots and tested in duplicate per ELISA plate on 3 different days in 4 different laboratories (Bristol, Ghent, Hannover, Uppsala).

Development and laboratory validation of a lateral flow device for the detection of foot-and-mouth disease virus in clinical samples.

A lateral flow device (LFD) for the detection of all seven serotypes of foot-and-mouth disease virus (FMDV) was developed using a monoclonal antibody (Mab 1F10) shown to be pan-reactive to FMDV strains of each serotype by ELISA. The performance of the LFD was evaluated in the laboratory on suspensions of vesicular epithelia (304 positive and 1003 negative samples) from suspected cases of vesicular disease collected from 86 countries between 1965 and 2008 and negative samples collected from healthy animals. The diagnostic sensitivity of the LFD for FMDV was similar at 84% compared to 85% obtained by the reference method of antigen ELISA, and the diagnostic specificity of the LFD was approximately 99% compared to 99.

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.

Detection of individual microbial pathogens by proximity ligation.

Background: Nucleic acid amplification allows the detection of single infectious agents. Protein-based assays, although they provide information on ongoing infections, have substantially less detection sensitivity.

Methods: We used proximity ligation reactions to detect proteins on bacteria and virus particles via nucleic acid amplification.

Cytosolic thymidine kinase is a specific histopathologic tumour marker for breast carcinomas.

Thymidine kinase 1 (TK1), an enzyme involved in the synthesis of precursors for DNA, and thus proliferation dependent, has been suggested as a good tumour marker. We have recently developed poly/monoclonal antibodies against TK1, which proved useful for diagnostics in both serum and immunohistochemistry of cancer patients. The anti-TK1 monoclonal antibodies (mAbs) 1D11 and 1E3 were characterized by Western blot, immunoprecipitation and flow cytometry.

Prevalence of equine herpesvirus types 2 and 5 in horse populations by using type-specific PCR assays.

Equineherpesvirustypes 2 and 5 (EHV-2andEHV-5)have a rather unclearpathogenicity and distribution within the equid population. In order to gain more information on the prevalence of these two viruses, type-specific PCR assays were developed to detect viral DNA in nasal specimens and in peripheral blood leukocytes (PBLs) of adult horses and foals from various regions of Europe, i.e.