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1.
J Vet Med Sci ; 83(11): 1722-1725, 2021 Nov 16.
Article in English | MEDLINE | ID: covidwho-1518345

ABSTRACT

We investigated the seroprevalence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) among dogs in the Tokyo area via enzyme-linked immunosorbent assay (ELISA) using the spike protein as the target antigen. Plasma samples from 494 household dogs and blood-donor dogs were tested from July 2020 to January 2021. Of these samples, three showed optical densities that were higher than the mean plus two standard deviations of the mean of the negative-control optical densities (ODs). Of these three samples, only the sample with the highest OD by ELISA was confirmed positive by virus neutralization testing. The positive dog presented no SARS-CoV-2-related symptoms. The positivity rate of SARS-CoV-2 infections among dogs in the Tokyo area was approximately 0.2%.


Subject(s)
COVID-19 , Dog Diseases , Animals , Antibodies, Viral , COVID-19/veterinary , Dog Diseases/epidemiology , Dogs , Enzyme-Linked Immunosorbent Assay/veterinary , Japan/epidemiology , SARS-CoV-2 , Seroepidemiologic Studies , Spike Glycoprotein, Coronavirus
2.
J Vet Diagn Invest ; 33(5): 969-974, 2021 Sep.
Article in English | MEDLINE | ID: covidwho-1298415

ABSTRACT

Avian influenza A(H5) viruses (avian IAVs) pose a major threat to the economy and public health. We developed an antigen-ELISA (ag-ELISA) and a colloidal gold-based immunochromatographic strip for the rapid detection of avian A(H5) viruses. Both detection methods displayed no cross-reactivity with other viruses (e.g., other avian IAVs, infectious bursal disease virus, Newcastle disease virus, infectious bronchitis virus, avian paramyxovirus). The ag-ELISA was sensitive down to 0.5 hemagglutinin (HA) units/100 µL of avian A(H5) viruses and 7.5 ng/mL of purified H5 HA proteins. The immunochromatographic strip was sensitive down to 1 HA unit/100 µL of avian A(H5) viruses. Both detection methods exhibited good reproducibility with CVs < 10%. For 200 random poultry samples, the sensitivity and specificity of the ag-ELISA were 92.6% and 98.8%, respectively, and for test strips were 88.9% and 98.3%, respectively. Both detection methods displayed high specificity, sensitivity, and stability, making them suitable for rapid detection and field investigation of avian A(H5) viruses.


Subject(s)
Infectious bronchitis virus , Influenza in Birds , Animals , Antibodies, Monoclonal , Antibodies, Viral , Chickens , Enzyme-Linked Immunosorbent Assay/veterinary , Gold Colloid , Influenza in Birds/diagnosis , Reproducibility of Results , Sensitivity and Specificity
3.
Virulence ; 12(1): 1597-1609, 2021 12.
Article in English | MEDLINE | ID: covidwho-1268053

ABSTRACT

The COVID-19 pandemic caused by the coronavirus SARS-CoV-2 is continuing to spread globally. SARS-CoV-2 infections of feline and canine species have also been reported. However, it is not entirely clear to what extent natural SARS-CoV-2 infection of pet dogs and cats is in households. We have developed enzyme-linked immunosorbent assays (ELISAs) using recombinant SARS-CoV-2 nucleocapsid (N) protein and the receptor-binding-domain (RBD) of the spike protein, and the SARS-CoV-2 spike-pseudotyped vesicular stomatitis virus (VSV)-based neutralization assay to screen serum samples of 239 pet cats and 510 pet dogs in Minnesota in the early phase of the COVID-19 pandemic from mid-April to early June 2020 for evidence of SARS-CoV-2 exposures. A cutoff value was used to identify the seropositive samples in each experiment. The average seroprevalence of N- and RBD-specific antibodies in pet cats were 8% and 3%, respectively. Among nineteen (19) N-seropositive cat sera, fifteen (15) exhibited neutralizing activity and seven (7) were also RBD-seropositive. The N-based ELISA is also specific and does not cross react with antigens of common feline coronaviruses. In contrast, SARS-CoV-2 antibodies were detected at a very low percentage in pet dogs (~ 1%) and were limited to IgG antibodies against SARS-CoV-2 N protein with no neutralizing activities. Our results demonstrate that SARS-CoV-2 seropositive rates are higher in pet cats than in pet dogs in MN early in the pandemic and that SARS-CoV-2 N-specific IgG antibodies can detect SARS-CoV-2 infections in companion animals with higher levels of specificity and sensitivity than RBD-specific IgG antibodies in ELISA-based assays.


Subject(s)
COVID-19 Serological Testing/veterinary , COVID-19/veterinary , Pets/virology , SARS-CoV-2/isolation & purification , Animals , Antibodies, Neutralizing/blood , Antibodies, Viral/blood , COVID-19/diagnosis , COVID-19/epidemiology , Cats , Coronavirus Nucleocapsid Proteins/immunology , Coronavirus, Feline/immunology , Coronavirus, Feline/isolation & purification , Dogs , Enzyme-Linked Immunosorbent Assay/veterinary , Minnesota/epidemiology , Phosphoproteins/immunology , SARS-CoV-2/immunology , Sensitivity and Specificity , Seroepidemiologic Studies , Spike Glycoprotein, Coronavirus/immunology
4.
Transbound Emerg Dis ; 68(4): 1779-1785, 2021 Jul.
Article in English | MEDLINE | ID: covidwho-944802

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic with millions of infected humans and hundreds of thousands of fatalities. As the novel disease - referred to as COVID-19 - unfolded, occasional anthropozoonotic infections of animals by owners or caretakers were reported in dogs, felid species and farmed mink. Further species were shown to be susceptible under experimental conditions. The extent of natural infections of animals, however, is still largely unknown. Serological methods will be useful tools for tracing SARS-CoV-2 infections in animals once test systems are evaluated for use in different species. Here, we developed an indirect multi-species ELISA based on the receptor-binding domain (RBD) of SARS-CoV-2. The newly established ELISA was evaluated using 59 sera of infected or vaccinated animals, including ferrets, raccoon dogs, hamsters, rabbits, chickens, cattle and a cat, and a total of 220 antibody-negative sera of the same animal species. Overall, a diagnostic specificity of 100.0% and sensitivity of 98.31% were achieved, and the functionality with every species included in this study could be demonstrated. Hence, a versatile and reliable ELISA protocol was established that enables high-throughput antibody detection in a broad range of animal species, which may be used for outbreak investigations, to assess the seroprevalence in susceptible species or to screen for reservoir or intermediate hosts.


Subject(s)
COVID-19 , Cat Diseases , Cattle Diseases , Rodent Diseases , Animals , Antibodies, Viral , COVID-19/veterinary , Cat Diseases/virology , Cats , Cattle , Cattle Diseases/virology , Chickens , Enzyme-Linked Immunosorbent Assay/veterinary , Ferrets , Humans , Mice , Rabbits , Rodent Diseases/virology , SARS-CoV-2 , Seroepidemiologic Studies
5.
J Vet Diagn Invest ; 32(4): 585-588, 2020 Jul.
Article in English | MEDLINE | ID: covidwho-603625

ABSTRACT

Influenza D virus (IDV) is considered a new agent involved in bovine respiratory disease (BRD). Based on seroprevalence studies or isolation from clinical samples, this virus has been detected on several continents and in several animal species, including cattle, pigs, camel, horses, and goats. We used an indirect in-house ELISA to detect anti-IDV antibodies in 165 serum samples from bulls on 116 farms in the province of La Pampa, Argentina. Eighty-five of 116 (73%) farms had at least 1 positive animal, and 112 of 165 (68%) of the analyzed samples were positive. There were no significant differences in the proportion of seropositive samples depending on the geographic region in which the samples were taken. Our results suggest that IDV infection is endemic in La Pampa; the clinical importance of IDV in Argentina remains to be investigated.


Subject(s)
Cattle Diseases/epidemiology , Orthomyxoviridae Infections/veterinary , Thogotovirus/isolation & purification , Animals , Antibodies, Viral/blood , Argentina/epidemiology , Cattle , Cattle Diseases/virology , Enzyme-Linked Immunosorbent Assay/veterinary , Male , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Prevalence , Seroepidemiologic Studies
6.
J Immunol Methods ; 483: 112808, 2020 08.
Article in English | MEDLINE | ID: covidwho-602160

ABSTRACT

Three different porcine enteric coronaviruses (PECs), i.e., porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV) and porcine Deltacoronavirus (PDCoV) are currently circulating in U.S. commercial swine herds. Differential diagnosis of PECs relies on laboratory methods. This study describes the development of an ELISA-like multiplex planar immunoassay based on virus-specific recombinant S1 proteins printed in an array of spots at the bottom of a 96-well microplate for simultaneous detection differential serodiagnosis of PEDV, TGEV, PDCoV in a single sample. The technology overall format and working principle is similar to the solid-phase standard ELISA. After the three typical incubation steps, the reaction was visualized as blue spots which intensity correlated with antibody levels to specific viral antigen target in the array. The diagnostic performance of the assay was evaluated on known status serum samples (n = 480) collected over time (day post-inoculation -7, 0, 7, 14, 21, 28, 35, and 42) from pigs inoculated with PEDV, TGEV Purdue, TGEV Miller, PDCoV (USA/IL/2014), or mock inoculated with culture media under experimental conditions. Antigen-specific cut-offs were selected to ensure 100% diagnostic and analytical specificity for each given antigen target. The overall diagnostic sensitivity was 92% (44/48 positives, 95% confidence interval (CI) 98,100) for PEDV S1, 100% (95/95 positives, 95% CI 98, 100) for TGEV S1, and 98% (47/48 positives, 95% CI 97, 100) for PDCoV S1. The results of this study demonstrate that the AgroDiag PEC multiplex immunoassay is an efficient and reliable test for differential detection and serodiagnosis of PEDV, TGEV and PDCoV.


Subject(s)
Alphacoronavirus/immunology , Antibodies, Viral/blood , Coronavirus Infections/veterinary , Enzyme-Linked Immunosorbent Assay/veterinary , Immunoglobulin G/blood , Serologic Tests/veterinary , Animals , Biomarkers/blood , Coronavirus Infections/diagnosis , Coronavirus Infections/immunology , Coronavirus Infections/virology , Deltacoronavirus/immunology , Diagnosis, Differential , Gastroenteritis, Transmissible, of Swine/diagnosis , Gastroenteritis, Transmissible, of Swine/immunology , Gastroenteritis, Transmissible, of Swine/virology , Porcine epidemic diarrhea virus/immunology , Predictive Value of Tests , Reproducibility of Results , Swine , Transmissible gastroenteritis virus/immunology
7.
Transbound Emerg Dis ; 67(4): 1745-1749, 2020 Jul.
Article in English | MEDLINE | ID: covidwho-71843

ABSTRACT

The pandemic SARS-CoV-2 has been reported in 123 countries with more than 5,000 patients died from it. However, the original and intermediate hosts of the virus remain unknown. In this study, 1,914 serum samples from 35 animal species were used for detection of SARS-CoV-2-specific antibodies using double-antigen sandwich ELISA after validating its specificity and sensitivity. The results showed that no SARS-CoV-2-specific antibodies were detected in above samples which excluded the possibility of 35 animal species as intermediate host for SARS-CoV-2. More importantly, companion animals including pet dogs (including one dog the SARS-CoV-2 patient kept and two dogs which had close contact with it) and cats, street dogs and cats also showed serological negative to SARS-CoV-2, which relieved the public concerns for the pets as SARS-CoV-2 carriers.


Subject(s)
Clinical Laboratory Techniques/veterinary , Coronavirus Infections/epidemiology , Pandemics , Pneumonia, Viral/epidemiology , Animals , Animals, Wild , Antibodies, Viral/blood , Betacoronavirus/immunology , COVID-19 , COVID-19 Testing , Cats , Coronavirus Infections/diagnosis , Coronavirus Infections/veterinary , Dogs , Enzyme-Linked Immunosorbent Assay/veterinary , Pets , SARS-CoV-2
8.
N Z Vet J ; 68(1): 54-59, 2020 Jan.
Article in English | MEDLINE | ID: covidwho-3840

ABSTRACT

Aims: To determine the seroprevalence of canine respiratory coronavirus (CRCoV) in New Zealand dogs, and to explore associations with age, sex, breed, month, and geographical region of sampling and reported presence of clinical signs suggestive of respiratory disease.Methods: A total of 1,015 canine serum samples were randomly selected from submissions to a diagnostic laboratory between March and December 2014, and were analysed for CRCoV antibodies using a competitive ELISA. Logistic regression analysis was used to determine associations between seroprevalence of CRCoV and breed category, age, sex, sampling month, region, and reported health status of dogs.Results: Overall, 538/1,015 (53.0%) samples were seropositive for CRCoV, with 492/921 (53.4%) positive dogs in the North Island and 46/94 (49%) in the South Island. Age of dog, sampling month, region, and presence of abnormal respiratory signs were included in the initial logistic regression model. Seroprevalence was higher in dogs aged ≥3 compared with ≤2 years (p < 0.01). The lowest seroprevalence was observed in July (30/105; 28.5%) and August (32/100; 32%), and the highest in June (74/100; 74%). Seroprevalence in dogs from Auckland was higher than in dogs from the Hawkes Bay, Manawatu, Marlborough, and Waikato regions (p < 0.05). Abnormal respiratory signs (coughing, nasal discharge, or sneezing) were reported for 28/1,015 (2.8%) dogs sampled. Seroprevalence for CRCoV tended to be higher among dogs with respiratory signs (67.9 (95% CI = 47.6-83.4)%) than dogs with no reported respiratory signs (52.6 (95% CI = 49.5-55.7)%).Conclusions: Serological evidence of infection with CRCoV was present in more than half of the dogs tested from throughout New Zealand. Differences in CRCoV seroprevalence between regions and lack of seasonal pattern indicate that factors other than external temperatures may be important in the epidemiology of CRCoV in New Zealand.Clinical relevance: Our data suggest that CRCoV should be included in investigations of cases of infectious canine tracheobronchitis, particularly if these occur among dogs vaccinated with current vaccines, which do not include CRCoV antigens.


Subject(s)
Antibodies, Viral/blood , Coronavirus Infections/veterinary , Coronavirus, Canine/immunology , Dog Diseases/epidemiology , Animals , Coronavirus Infections/blood , Coronavirus Infections/epidemiology , Coronavirus Infections/virology , Coronavirus, Canine/isolation & purification , Dog Diseases/blood , Dog Diseases/virology , Dogs , Enzyme-Linked Immunosorbent Assay/veterinary , Logistic Models , New Zealand/epidemiology , Seroepidemiologic Studies
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