scholarly journals Evaluation of a Commercial Competitive ELISA Test Kit for the Detection of Group-Specific Antibodies to Bluetongue Virus

1993 ◽  
Vol 5 (3) ◽  
pp. 336-340 ◽  
Author(s):  
A. Afshar ◽  
H. C. Trotter ◽  
G. C. Dulac ◽  
J. J. Reddington
Keyword(s):  
Bluetongue Virus ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Test Kit ◽  
Field Samples ◽  
The Usa ◽  
Haemorrhagic Disease

The performance of a competitive (c) enzyme-linked immunosorbent assay (ELISA) test kit, Blueplate Special@ (BPS), commercially produced by DiagXotics (Wilton, CT) for detection of group-specific antibodies to bluetongue virus (BTV) was compared with that of an internationally endorsed cELISA-I. A total of 1,026 serum samples were tested in this study: 133 samples from 23 calves and 3 sheep experimentally infected with South African isolates of 19 BTV serotypes and US isolates of 5 BTV serotypes and 7 calves infected with US isolates of 2 epizootic haemorrhagic disease of deer virus (EHDV); 102 paired sera from cattle, sheep, and goats experimentally infected with the Australian isolates of BTV, EHDV, and Palyam virus; 229 bovine and ovine samples of Canadian origin (BTV free); and 562 bovine and ovine field samples from the USA and Barbados (BTV endemic). Seroconversion was demonstrable by the BPS cELISA 10 days postinfection in all experimental animals inoculated with BTV, with the exception of 4 calves in which there was a delay of 10–20 days. Similar to the cELISA-I, none of the sera from calves inoculated with US and Australian isolates of EHDV and Palyam viruses cross-reacted with the BTV antigen in the BPS cELISA. The total agreement between the two assays for all the total bovine and ovine field sera was 98.1%. The overall results substantiate the usefulness of the BPS cELISA test kit for monitoring animal sera for group-specific antibodies to BTV. The slightly lower analytical sensitivity associated with the detection of antibody during early phase of infection in some animals would not be significant in the context of herd testing or any regulatory program.

scholarly journals PEDV Infection Generates Conformation-Specific Antibodies That Can Be Effectively Detected by a Cell-Based ELISA

Viruses ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 303
Author(s):  
Wei-Ting Hsu ◽  
Chia-Yu Chang ◽  
Chih-Hsuan Tsai ◽  
Sung-Chan Wei ◽  
Huei-Ru Lo ◽  
...  
Keyword(s):  
Recombinant Baculovirus ◽  
Immunocytochemical Staining ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Perfect Agreement ◽  
Specific Antibodies ◽  
Diarrhea Virus ◽  
Serological Studies ◽  
A Cell ◽  
Infected Insect

Porcine epidemic diarrhea virus (PEDV) is a coronavirus that causes serious and highly contagious enteric disease in swine worldwide. In this study, we constructed a recombinant baculovirus (S-Bac) expressing full-length spike protein of the virulent epidemic genotype 2b (G2b) PEDV strain for serological studies of infected pigs. We found that most spike-specific antibodies produced upon PEDV infection in pigs are conformation-specific and they could be detected on S-Bac-infected insect cells by immunofluorescent assay, but they were insensitive to Western blot analysis, the typical method for antiserum analysis. These results indicated that spike conformation is crucial for serum recognition. Since it is difficult to purify trimeric spike membrane protein for conventional enzyme-linked immunosorbent assay (ELISA), we used S-Bac to generate a novel cell-based ELISA for convenient PEDV detection. We analyzed 100 pig serum samples, and our cell-based ELISA exhibited a sensitivity of 100%, a specificity of 97%, and almost perfect agreement [Cohen’s kappa coefficient value (κ) = 0.98] with immunocytochemical staining results. Our cell-based ELISA rapidly presented antigen for proper detection of conformation-specific antibodies, making PEDV detection more convenient, and it will be useful for detecting many viral diseases in the future.

scholarly journals Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparision of the assay with ELISA

1992 ◽  
Vol 34 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Eide Dias Camargo ◽  
Paulo Mutuko Nakamura ◽  
Adelaide José Vaz ◽  
Marcos Vinícius da Silva ◽  
Pedro Paulo Chieffi ◽  
...  
Keyword(s):  
Low Cost ◽  
Clinical Signs ◽  
Toxocara Canis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Normal Individuals ◽  
Before And After ◽  
Dot Elisa ◽  
Stability Time

The dot-enzyme-linked immunosorbent assay (dot-ELISA) was standardized using somatic (S) and excretory-secretory (ES) antigens of Toxocara-canis for the detection of specific antibodies in 22 serum samples from children aged 1 to 15 years, with clinical signs of toxocariasis. Fourteen serum samples from apparently normal individuals and 28 sera from patients with other pathologies were used as controls. All samples were used before and after absorption with Ascaris suum extract. When the results were evaluated in comparison with ELISA, the two tests were found to have similar sensitivity, but dot-ELISA was found to be more specific in the presence of the two antigens studied. Dot-ELISA proved to be effective for the diagnosis of human toxocariasis, presenting advantages in terms of yield, stability, time and ease of execution and low cost.

scholarly journals Evaluation of Vaccination Strategy Against Rabies in Hong Kong Macaques

2021 ◽  
Author(s):  
Omid Nekouei ◽  
Paolo Martelli ◽  
Sophie St-Hilaire ◽  
Hui Suk Wai ◽  
Karthiyani Krishnasamy ◽  
...  
Keyword(s):  
Hong Kong ◽  
Vaccination Strategy ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Response ◽  
Test Kit ◽  
In The Wild ◽  
The Government ◽  
Specific Association ◽  
Antibody Levels

Rabies is a fatal zoonotic disease that can affect all mammals. Following the directives of the rabies ordinance of the Government of Hong Kong, all wild macaques captured under an ongoing sterilization program (since 2000) were vaccinated against rabies. The main objective of this study was to assess the serological response to rabies vaccination in the population of Hong Kong macaques. An inactivated rabies vaccine was subcutaneously administered to captured macaques under anesthesia. In a 2015 field survey, blood samples from the animals were collected and stored in -80℃ freezer. In July 2021, all frozen sera from vaccinated animals were prepared and tested for antibodies against rabies virus using a commercial enzyme-linked immunosorbent assay (ELISA) test. The test results were dichotomized at the recommended cut-off point of the test kit. Sixty-five samples from the vaccinated macaques were available for this study. All of these animals had received at least one dose of vaccine (1 vaccination) between 2008 and 2015. The interval between the 1 vaccination and blood sampling dates ranged from 21 to 2,779 days. Only five of the 65 macaques had a second vaccination record at the time of sampling; all five had high antibody levels. Among the remaining macaques, 77% (46/60) were positive for rabies antibodies. No specific association was observed between the post-vaccination period and the antibody titer of these macaques and no adverse reactions to vaccination were reported. The current vaccination strategy in Hong Kong macaques appears to effectively elicit rabies antibodies in a high proportion of macaque populations in the wild (78-87%). However, reaching the precise level of protection against a potential challenge with the virus should further be investigated.

scholarly journals Investigation ofAnaplasma marginaleSeroprevalence in a Traditionally Managed Large California Beef Herd

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Thomas R. Tucker ◽  
Sharif S. Aly ◽  
John Maas ◽  
Josh S. Davy ◽  
Janet E. Foley
Keyword(s):  
Anaplasma Marginale ◽  
Incidence Density ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Cattle Disease ◽  
Angus Cattle ◽  
Increased Risk ◽  
Test Kit ◽  
Incidence Density Rate ◽  
Density Rate

Recent observations by stakeholders suggested that ecosystem changes may be driving an increased incidence of bovine erythrocytic anaplasmosis, resulting in a reemerging cattle disease in California. The objective of this prospective cohort study was to estimate the incidence ofAnaplasma marginaleinfection using seroconversion in a northern California beef cattle herd. A total of 143 Black Angus cattle (106 prebreeding heifers and 37 cows) were enrolled in the study. Serum samples were collected to determineAnaplasma marginaleseroprevalence using a commercially available competitive enzyme-linked immunosorbent assay test kit. Repeat sampling was performed in seronegative animals to determine the incidence density rate from March through September (2013). Seroprevalence of heifers was significantly lower than that of cows at the beginning of the study (P<0.001) but not at study completion (P=0.075). Incidence density rate ofAnaplasma marginaleinfection was 8.17 (95% confidence interval: 6.04, 10.81) cases per 1000 cow-days during the study period. Study cattle becameAnaplasma marginaleseropositive and likely carriers protected from severe clinical disease that might have occurred had they been first infected as mature adults. No evidence was found within this herd to suggest increased risk for clinical bovine erythrocytic anaplasmosis.

scholarly journals Detection of avian reovirus antibodies in layer birds of small scale commercial farms in Dinajpur district of Bangladesh

2016 ◽  
Vol 1 (3) ◽  
pp. 612-621 ◽  
Author(s):  
Abdus Salam ◽  
Md Atiqul Haque ◽  
Md Mostafizer Rahman ◽  
Mir Rowshan Akter ◽  
Farzana Afroz
Keyword(s):  
Antibody Titer ◽  
Specific Antibody ◽  
Age Groups ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Small Scale ◽  
Avian Reovirus ◽  
Serum Samples ◽  
Aged Group ◽  
Test Kit

The present study was conducted on layer birds of different age groups to determine specific antibody titer level against avian reovirus (ARV) by indirect enzyme linked immunosorbent assay (iELISA) at Dinajpur district of Bangladesh. This study showed that ARV specific antibody positive cases were 84 out of 90 blood serum samples and the highest antibody titer was 26120 and lowest antibody titer was 288. The total 93.33% sera samples were showed positive result. The study showed that 100% sera sample were positive against ARV at 6 weeks of aged group and the highest, lowest and mean antibody titer were 13917, 4895 and 10269 respectively. On the other hand 88.88% sera sample were positive against ARV at 10 weeks of aged group and the highest, lowest and mean antibody titer were 9779, 288 and 5689.89 respectively. The sera sample collected from 14 weeks of aged group showed 88.88% positive and the highest, lowest and mean antibody titer were 11727, 871 and 5250 respectively. The sera sample collected from 18 weeks of aged group showed 88.88% positive against ARV and the highest, lowest and mean antibody titer were 24440, 1234 and 12648.89 respectively. The sera sample collected from 22 weeks of aged group were 100% positive against ARV and the highest, lowest and mean antibody titer were 26120, 1752 and 11373.89 respectively. The sera sample collected from 26 weeks of aged group showed 100% positive against ARV and the highest, lowest and mean antibody titer were 8566, 1630 and 4327.44 respectively. The sera sample collected from 30 weeks of aged group showed 100% positive against ARV and the highest, lowest and mean antibody titer were 13431, 1989 and 5890.56 respectively. The sera sample collected from 40 weeks of aged group showed 77.77% positive against ARV and the highest, lowest and mean antibody titer were 14618, 433 and 5103.22 respectively. The sera sample collected from 48 weeks of aged group showed 88.88% positive against ARV and the highest, lowest and mean antibody titer were 14553, 957 and 7436.5 respectively. In conclusion it is evident that avian reovirus-specific antibody was successfully detected through commercially available avian reovirus antibody test kit (ELISA kit) and the virus induced a significant antibody titer indicating the affecting virus was absolutely ARV.Asian J. Med. Biol. Res. December 2015, 1(3): 612-621

scholarly journals Detection of bluetongue virus antibodies in sheep from Paraná, Brazil

2020 ◽  
Vol 41 (3) ◽  
pp. 879
Author(s):  
Maria Carolina Ricciardi Sbizera ◽  
Luiz Fernando Coelho da Cunha Filho ◽  
Michele Lunardi ◽  
Simone Fernanda Nedel Pertile ◽  
Thais Helena Constantino Patelli ◽  
...  
Keyword(s):  
Bluetongue Virus ◽  
Animal Health ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Results ◽  
Serum Samples ◽  
Agar Gel ◽  
Serological Tests ◽  
High Occurrence ◽  
Predominant Factor ◽  
World Organization

Bluetongue (BT) is an infectious and non-contagious disease caused by bluetongue virus (BTV) belonging to the genus Orbivirus. It is transmitted by a hematophagous vector, Culicoides sp., to ruminants, particularly to sheep, which are most susceptible to this disease. The main serological tests are agar gel immunodiffusion (AGID), which is recommended by the World Organization for Animal Health (OIE), and the competitive enzyme-linked immunosorbent assay (cELISA), which has the advantage of no cross-reaction with other orbiviruses. The aim was to compare the results of these two tests by conducting them on sera collected from sheep in the state of Paraná, Brazil. From March to October 2017, serum samples were collected from 270 sheep from 10 farms in six mesoregions of Paraná. The samples were subjected to AGID and cELISA to detect antibodies against BTV. Based on the test results, we classified the sheep as low, moderate, and high occurrence. The results demonstrated that 64.81% (175/270) of the sheep were seropositive through the cELISA test, showing a high occurrence, and 41.11% (111/270) were seropositive through the AGID test, indicating a moderate occurrence. The concordance between the tests was moderate (0.51) as determined by the Kappa coefficient. Among the studied farms, 90% (9/10) presented at least one seropositive sheep, and the number of animals tested positive by the cELISA test was higher than those by the AGID test. Favorable climate, which favors the presence and multiplication of the culicoid vector and the occurrence of infection, was the biggest predominant factor responsible for the obtained results. The low occurrence in farms with milder climate suggest that the presence of antibodies also occurs due to the low pathogenicity of circulating serotypes in the different mesoregions studied. It is concluded that BTV infection is present in the sheep herds in Paraná, and the occurrence was moderate detected by AGID test and high detected by cELISA test.

DETECTION OF SPECIFIC ANTIBODIES TO THE NUCLEOCAPSID PROTEIN FRAGMENTS OF SEVERE ACUTE RESPIRATORY SYNDROME-CORONAVIRUS AND SCOTOPHILUS BAT CORONAVIRUS-512 IN THREE INSECTIVOROUS BAT SPECIES

2018 ◽  
Vol 44 (04) ◽  
pp. 179-188 ◽  
Author(s):  
Yi-Ning Chen ◽  
Bo-Gang Su ◽  
Hung-Chang Chen ◽  
Cheng-Han Chou ◽  
Hsi-Chi Cheng
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Nucleocapsid Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Protein Fragments ◽  
Specific Antibodies ◽  
History Of ◽  
Polymerase Chain ◽  
Carboxyl Terminal ◽  
Bat Coronavirus

Bats are the natural reservoirs of severe acute respiratory syndrome-coronavirus (SARS-CoV). Six Alphacoronavirus and five Betacoronavirus have been detected in many bat species, including SARS-related CoV and Middle East respiratory syndrome (MERS)-related CoV. In Taiwan, SARS-related CoV, belonging to Betacoronavirus, has been detected in Rhinolophus monoceros. Scotophilus bat CoV-512, belonging to Alphacoronavirus, has been detected in Scotophilus kuhlii, Miniopterus fuliginosus, and Rhinolophus monoceros by using reverse transcription polymerase chain reaction (RT-PCR). To understand the infection history of CoV in these three insectivorous bat populations, CoV-specific antibodies were surveyed by using western blot (WB) analysis and indirect enzyme-linked immunosorbent assay (ELISA). The carboxyl terminal fragment of nucleocapsid protein (N3) of SARS-CoV and Scotophilus bat CoV-512 were used as the antigen in the assays. Of the 52 serum samples obtained from Scotophilus kuhlii, 29 samples (56%) were tested positive for Scotophilus bat CoV-512-specific antibodies through ELISA. Of the 63 serum samples obtained from Rhinolophus monoceros, 9 samples were tested positive for only SARS-CoV-specific antibodies, 7 samples were tested positive for only Scotophilus bat CoV-512-specific antibodies, and 16 samples (25.4%) were tested positive for both antibodies through WB analysis. Only 1 of 18 Miniopterus bat serum samples tested positive for Scotophilus bat CoV-512-specific antibodies through ELISA. Lactating female bats had higher positive rates of CoV-specific antibodies than non-lactating female and male bats did. Our findings were crucial for understanding CoV infection history in three insectivorous bat species and important for the control of bat-borne zoonosis diseases.

scholarly journals Variations in the detection of anti-PEDV antibodies in serum samples using three diagnostic tests – short communication

2018 ◽  
Vol 66 (2) ◽  
pp. 337-342 ◽  
Author(s):  
Jan Plut ◽  
Ivan Toplak ◽  
Marina Štukelj
Keyword(s):  
Clinical Signs ◽  
Elisa Test ◽  
Diagnostic Methods ◽  
P Value ◽  
Serum Samples ◽  
Blocking Elisa ◽  
Test Kit ◽  
Two Samples ◽  
Antibody Tests ◽  
Higher Sensitivity

Over the last few years several porcine epidemic diarrhoea (PED) outbreaks have been discovered in Europe including the first PED case in Slovenia in January 2015. The aim of this study was to determine when PED virus (PEDV) infection started in Slovenia. Serum samples collected between 2012 and 2016 were tested. Three hundred and seventy-five serum samples were collected from 132 Slovenian small, one-site pig farms. Samples were tested for PEDV antibodies utilising three different serological methods: commercially-available indirect ELISA, in-house blocking ELISA test and Immunoperoxidase Monolayer Assay (IPMA) test. One hundred and seventy (45.33%) tested samples were found positive by the commercially-available ELISA test kit, and 10 (5.68%) of these 170 samples found positive were positive by the in-house blocking ELISA. Only these 10 samples were collected from a farm where clinical signs of PED infection had been observed and PEDV was confirmed by RT-PCR methodology; the other 160 samples were collected randomly. Thirty-two samples with the highest S/P value obtained with the commercial ELISA were all negative with IPMA. Reasons for the high variance in the results obtained remain unclear; more research is required to ensure higher sensitivity and specificity in terms of PEDV antibody tests and other PED diagnostic methods.

Evaluation of a saliva test kit for feline leukemia virus antigen

1996 ◽  
Vol 32 (5) ◽  
pp. 397-400
Author(s):  
SD Babyak ◽  
MG Groves ◽  
DS Dimski ◽  
J Taboada
Keyword(s):  
Leukemia Virus ◽  
Virus Antigen ◽  
Elisa Test ◽  
Feline Leukemia Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Kit ◽  
Test Kit ◽  
Blood Smears ◽  
Indirect Immunofluorescent ◽  
Saliva Test

An enzyme-linked immunosorbent assay (ELISA) test kit for the detection of feline leukemia virus (FeLV) antigen in saliva was evaluated in 150 cats. Saliva and blood samples from all cats were tested for FeLV using the saliva ELISA kit and a plasma ELISA kit, respectively. These results were compared with indirect immunofluorescent antibody (IFA) testing of blood smears also obtained from each cat. The proportion of cats that tested positive were 10%, 7%, and 8% for each test, respectively. Using the IFA test as the gold standard, the saliva FeLV test had a sensitivity of 91.7% and specificity of 97.1%, while the plasma ELISA test had a sensitivity of 91.7% and specificity of 100%.

scholarly journals Development of an enzyme-linked immunosorbent assay (ELISA) for the detection of specific antibodies against an H7N7 and an H3N8 equine influenza virus

1984 ◽  
Vol 93 (3) ◽  
pp. 609-620 ◽  
Author(s):  
M. S. Denyer ◽  
J. R. Crowther ◽  
R. C. Wardley ◽  
R. Burrows
Keyword(s):  
Influenza Virus ◽  
Immunosorbent Assay ◽  
Solid Phase ◽  
Allantoic Fluid ◽  
Influenza Viruses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Equine Influenza Virus ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Equine Influenza

SummaryThis paper describes a solid-phase microtitre plate enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to equine influenza viruses. Using egg-grown influenza viruses as the antigens attached to the solid phase, crossreactions were observed between an H7N7 equine virus (designated A1) and an H3N8 equine influenza virus (designated A2) when untreated antisera were tested. Absorption of antisera with egg-grown A/Porcine/Shope/1/33 influenza virus eliminated cross-reactive antibodies so that specific detection of anti-equine influenza A1 or A2 antibodies was possible.Examination of horse sera following vaccination with A1 and/or A2 isolates showed that antibodies were produced against antigen associated with egg allantoic fluid as well as against virus. Such antibodies were eliminated following the absorption of antisera with porcine influenza virus. Results using sera from horses with known vaccination histories confirmed that the ELISA preferentially detected antibodies homologous to the antigen attached to the solid phase and methods to evaluate the current serological state of individual horses by relating the titres of specific antibodies against equine influenza A1 and A2 isolates are shown. This ELISA provides a simple and rapid method of assessing specific antibodies from horse sera and offers advantages over the ‘routine’ HI and SRH assessments since it gives high precision, is economical of reagents and has the capacity to handle large numbers of serum samples.

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