In South Korea, where avian infl uenza virus subtypes H3N2, H5N1, H6N1, and H9N2 circulate or have been detected, 3 genetically similar canine infl uenza virus (H3N2) strains of avian origin (A/canine/Korea/01/2007, A/canine/ Korea/02/2007, and A/canine/Korea/03/2007) were isolated from dogs exhibiting severe respiratory disease. To determine whether the novel canine infl uenza virus of avian origin was transmitted among dogs, we experimentally infected beagles with this infl uenza virus (H3N2) isolate. The beagles shed virus through nasal excretion, seroconverted, and became ill with severe necrotizing tracheobronchitis and bronchioalveolitis with accompanying clinical signs (e.g., high fever). Consistent with histologic observation of lung lesions, large amounts of avian infl uenza virus binding receptor (SAα 2,3-gal) were identifi ed in canine tracheal, bronchial, and bronchiolar epithelial cells, which suggests potential for direct transmission of avian infl uenza virus (H3N2) from poultry to dogs. Our data provide evidence that dogs may play a role in interspecies transmission and spread of infl uenza virus.

The spike (S) gene of the attenuated porcine epidemic diarrhea virus (PEDV) DR13 was cloned and sequenced to further explore the functions of wild type PEDV and attenuated PEDV. Sequencing revealed a single large ORF of 4,149 nucleotides encoding a protein of 1,382 amino acids with predicted M(r) of 151 kDa. The coding region of the S gene of attenuated PEDV DR13 had 20 nucleotide changes that appeared to be significant determinants of function in that they produced changes in its predicted amino acid sequence. Notably, attenuated PEDV DR13 has previously been found to exhibit reduced pathogenicity in pigs. The regions containing these 20 nucleotide changes may therefore be crucial for PEDV pathogenicity. The attenuated PEDV DR13 S protein contains 28 Asn-Xaa-Ser/Thr sequons, 21 asparagines that are predicted to be N-glycosylated and a stretch of highly hydrophobic residues at positions 1,327-1,347, which is predicted to form an alpha-helix and to function as a membrane anchor. One (from N to K at 378) of the changes in the deduced amino acid sequence destroyed N-linked glycosylation sites, while another change (from N to S at 114) created a new one at a different location. These alterations in N-linked glycosylation sites reflected 3 nucleotide changes, which were related to the above-mentioned nucleotide changes and are suggested to influence the pathogenicity of attenuated PEDV DR13. Attenuated PEDV DR13 has 96.5, 96.4, 96.1, 93.9, 93.5 and 96.6% DNA sequence identities with CV777, Br1/87, JS-2004-2, Spk1, Chinju99 and parent DR13, respectively. Likewise, it shares 95.7, 95.4, 95.6, 92.0, 91.6 and 95.7% identity with those genes at the deduced amino acid sequence level. Phylogenetic analysis suggested that attenuated PEDV DR13 is closely related to CV777, Br1/87, JS-2004-2 and parent DR13, rather than to Spk1 and Chinju99 and is especially close to the Chinese PEDV strain JS-2004-2.

We present here a rapid immunochromatographic assay for the detection of Middle East respiratory syndrome coronavirus (MERS-CoV) antigen in the nasal swabs of dromedary camels. The assay is based on the detection of MERS-CoV nucleocapsid protein in a short time frame using highly selective monoclonal antibodies at room temperature. The relative sensitivity and specificity of the assay were found to be 93.90% and 100%, respectively, compared to that of the UpE and open reading frame 1A (Orf1A) real-time reverse transcriptase PCR (RT-PCR). The results suggest that the assay developed here is a useful tool for the rapid diagnosis and epidemiological surveillance of MERS-CoV infection in dromedary camels. Middle East respiratory syndrome coronavirus (MERS-CoV) is a newly identified human coronavirus associated with severe pulmonary syndrome and renal failure in infected patients (1). To date, a total of 843 persons in 21 different countries have been infected by the virus, with a resulting 37.95% mortality rate (2). The current MERS-CoV outbreak investigations suggest that camels are a source of human infections. Nevertheless, the exact route of transmission from camels to humans remains unclear (3).MERS-CoV is primarily diagnosed using molecular techniques. These include real-time reverse transcriptase PCR (RT-PCR) (4, 5), reverse transcription-loop-mediated isothermal amplification (RT-LAMP) (6) and reverse transcription-recombinase polymerase amplification (RT-RTPA) (7). Moreover, several serological assays have been used to detect MERS-CoV or closely related viruses in seropositive camels. These are protein microarrays (8-10), a recombinant spike immunofluorescent assay (11, 12), indirect enzyme-linked immunosorbent assay (ELISA) (13), microneutralization, and spike pseudoparticle neutralization (14). However, none of the serological tests have provided proof of the precise presence of MERS-CoV in camels.Molecular tests are relatively expensive, not available in all laboratories, and are mainly used for confirmatory purposes. For the purpose of screening of large numbers of animals in a short period of time, molecular tests are considered impractical; therefore, a rapid, cheap, sensitive, and specific test is needed for the diagnosis of MERS-CoV in camels. Here, we report the development and validation of an immunochromatographic assay (ICA) for the rapid qualitative detection of MERS-CoV antigen in dromedary camels. The assay is based on the detection of MERS-CoV nucleocapsid protein by highly selective monoclonal antibodies. MATERIALS AND METHODSThis study was carried out in two phases during the period of August to October 2014. In the first phase, the ICA was developed at the BioNote laboratory (South Korea). In the second phase, the performance and validation of the ICA were carried out at the veterinary laboratories of the Abu Dhabi Food Control Authority (United Arab Emirates).Peptides and monoclonal antibody synthesis. At first, the hydrophilic regions of the nucleocapsid gene of MERS-CoV were analyzed b...

This study was performed to determine the feasibility of using whole serum to detect antibodies to canine parvovirus (CPV) under nonlaboratory conditions and to evaluate the performance characteristics of an immunochromatography assay kit. Precise detection of levels of antibody against CPV in puppies can be used to determine a vaccination schedule, because maternal antibodies frequently result in the failure of protective vaccination, and can also be used to determine the antibody levels of infected puppies. Several methods for the titration of CPV antibodies have been reported, including the hemagglutination inhibition (HI) assay, which is considered the "gold standard." These methods, however, require intricate and time-consuming procedures. In this study, a total of 386 serum specimens were tested. Compared to the HI assay, the rapid assay had a 97.1% sensitivity and a 76.6% specificity (with a cutoff HI titer of 1:80). This single-step assay could be performed rapidly and easily without special equipment. The kit provides a reliable method for detection of anti-CPV antibody where laboratory support and personnel are limited.