scholarly journals Development and Validation of A Competitive ELISA Based on Virus-Like Particles of Serotype Senecavirus A to Detect Serum Antibodies

2020 ◽  
Author(s):  
Manyuan Bai ◽  
Rui Wang ◽  
Shiqi Sun ◽  
Yun Zhang ◽  
Hu Dong ◽  
...  
Keyword(s):  
Expression System ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Virus Like Particles ◽  
Specificity And Sensitivity ◽  
Indirect Immunofluorescent Assay ◽  
Viral Particles ◽  
Test Kit ◽  
Development And Validation

Abstract Virus-like particles (VLPs) are high-priority antigens with highly ordered repetitive structures, which are similar to natural viral particles. We have developed a competitive enzyme-linked immunosorbent assay (cELISA) for detecting antibodies directed against Senecavirus A (SVA). Our assay utilizes SVA VLPs that were expressed and assembled in an Escherichia coli expression system as the coating antigens. VLPs have better safety and immunogenicity than intact viral particles or peptides. The VLP-based cELISA was used to test 342 serum samples collected from different pig farms, and the results showed that its specificity and sensitivity were 100% and 94%, respectively. The consistency rates of cELISA with the BIOSTONE AsurDx™ Senecavirus A (SVA) Antibody Test Kit and an indirect immunofluorescent assay were 90.0% and 94.2%, respectively. Therefore, this VLP-based cELISA can be effectively and reliably used for the detection and discrimination of SVA infection in serum samples.

scholarly journals Development and validation of a competitive ELISA based on virus-like particles of serotype Senecavirus A to detect serum antibodies

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Manyuan Bai ◽  
Rui Wang ◽  
Shiqi Sun ◽  
Yun Zhang ◽  
Hu Dong ◽  
...  
Keyword(s):  
Expression System ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Virus Like Particles ◽  
Specificity And Sensitivity ◽  
Indirect Immunofluorescent Assay ◽  
Viral Particles ◽  
Test Kit ◽  
Development And Validation

AbstractVirus-like particles (VLPs) are high-priority antigens with highly ordered repetitive structures, which are similar to natural viral particles. We have developed a competitive enzyme-linked immunosorbent assay (cELISA) for detecting antibodies directed against Senecavirus A (SVA). Our assay utilizes SVA VLPs that were expressed and assembled in an E. coli expression system as the coating antigens. VLPs have better safety and immunogenicity than intact viral particles or peptides. The VLPs-based cELISA was used to test 342 serum samples collected from different pig farms, and the results showed that its specificity and sensitivity were 100% and 94%, respectively. The consistency rates of cELISA with the BIOSTONE AsurDx™ Senecavirus A (SVA) Antibody Test Kit and an indirect immunofluorescent assay were 90.0% and 94.2%, respectively. Therefore, this VLPs-based cELISA can be effectively and reliably used for the detection and discrimination of SVA infection in serum samples.

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 Lipopolysaccharide-Based Enzyme-Linked Immunosorbent Assay for Experimental Use in Detection of Antibodies to Lawsonia intracellularis in Pigs

2005 ◽  
Vol 12 (6) ◽  
pp. 693-699 ◽  
Author(s):  
J. J. Kroll ◽  
M. A. Eichmeyer ◽  
M. L. Schaeffer ◽  
S. McOrist ◽  
D. L. Harris ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Experimental Studies ◽  
Antibody Test ◽  
Controlled Study ◽  
Future Research ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lawsonia Intracellularis ◽  
Serum Samples ◽  
Specificity And Sensitivity

ABSTRACT An enzyme-linked immunosorbent assay (ELISA) for Lawsonia intracellularis was developed and compared with a whole-cell antigen-based immunofluorescence antibody test (IFAT). The antigen-containing lipopolysaccharide (LPS) was derived from Percoll gradient purified cultures of L. intracellularis by using a modification of the Westphal hot phenol procedure. The antigen was bound directly to polystyrene 96-well microtiter plates, and the assay was performed in an indirect ELISA format. Specificity and sensitivity values based on 80 known positive and 80 known negative serum samples from controlled experimental trials were 93.7% and 88.7%, respectively. Serological results from a controlled L. intracellularis challenge exposure study confirmed the high specificity and sensitivity of this assay (100% and 99.5%, respectively). Comparisons between the LPS ELISA and the IFAT in detecting anti-Lawsonia antibodies in this controlled study revealed significantly more LPS ELISA-positive pigs than IFAT-positive pigs on days 21, 28, 35, and 42 (P = 0.003, 0.030, 0.002, and 0.006, respectively). This indirect ELISA (LPS ELISA) test is an improved method of detecting antibodies in pigs soon after exposure to L. intracellularis, regardless of isolate type (vaccine or wild type) in experimental studies. The LPS ELISA may be used as a tool to support future research trials on vaccine efficacy and to further understand the immune response induced by L. intracellularis.

scholarly journals Antibodies anti-trypanosomatides in domestic cats in Paraná: who is at highest risk of infection?

2018 ◽  
Vol 27 (2) ◽  
pp. 232-236 ◽  
Author(s):  
Andressa Maria Rorato Nascimento de Matos ◽  
Eloiza Teles Caldart ◽  
Fernanda Pinto Ferreira ◽  
Keila Clarine Monteiro ◽  
Marielen de Souza ◽  
...  
Keyword(s):  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Domestic Cats ◽  
Serum Samples ◽  
Animal Group ◽  
Animal Protection ◽  
Leishmania Spp ◽  
In Series ◽  
Different Populations ◽  
Simple Logistic

Abstract The aim of this study were to detect antibodies anti-Leishmania spp. and anti-Trypanosoma cruzi in two different populations of domestic cats (Felis catus domesticus) from North Paraná referred for surgical castration and to determine which characteristics of the animals studied may be associated with seropositivity. Serum samples from 679 cats were analyzed using enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence antibody test (IFAT) in series. Associations between age, sex, race, year of care and animal group were verified using the simple logistic regression. Percentage of 8.5% (58/679) of cats were positive for Leishmania spp. and 7.6% (51/673) for T. cruzi by the tests ELISA and IFAT. Animals collected by non-governmental animal protection organizations presented more seropositivity for Leishmania spp. (p<0.0001). Results shown that Leishmania spp. and T. cruzi are present in domestic cats in the northern part of the state of Paraná, as well as, owners of non-governmental animal protection organizations may be more exposed to leishmaniasis when compared to other animal owners evaluated in the present study.

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 An enzyme-linked immunosorbent assay for the detection of IgG antibodies against Babesia equi in horses

2004 ◽  
Vol 34 (5) ◽  
pp. 1525-1529 ◽  
Author(s):  
Cristiane Divan Baldani ◽  
Rosangela Zacarias Machado ◽  
Paulo de Tarso Landgraf Botteon ◽  
Felipe Santoro Takakura ◽  
Carlos Luiz Massard
Keyword(s):  
Immunosorbent Assay ◽  
Serological Test ◽  
Epidemiological Studies ◽  
Sao Paulo ◽  
Enzyme Linked Immunosorbent Assay ◽  
São Paulo ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Babesia Equi ◽  
Specificity And Sensitivity

A crude antigenic preparation of Babesia equi was used to develop and establish the suitability of an enzyme-linked immunosorbent assay (ELISA) for the detection of parasite carriers. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkboard titrations. The specificity and sensitivity of the ELISA were 100 %. A total of 90 serum samples were taken from horses from the Northeast region of São Paulo State and examined for diagnosis of equine B. equi infection by ELISA. Approximately 75% (n=67) of all the horses tested were found serologically positive for B. equi. These results suggest that the ELISA described may prove to be an appropriate serological test for epidemiological studies on B. equi infections in the field and that equine piroplasmosis is a cause for serious concern in the State of São Paulo, Brazil.

Characterization of recombinant norovirus virus-like particles and evaluation of their applicability to the investigation of norovirus removal performance in membrane filtration processes

2015 ◽  
Vol 16 (3) ◽  
pp. 737-745
Author(s):  
N. Shirasaki ◽  
T. Matsushita ◽  
Y. Matsui ◽  
K. Ohno
Keyword(s):  
Membrane Filtration ◽  
Expression System ◽  
Small Animal ◽  
Microscopic Analysis ◽  
Peptide Mass Fingerprinting ◽  
Enzyme Linked Immunosorbent Assay ◽  
Electron Microscopic ◽  
Virus Removal ◽  
Virus Like Particles ◽  
Peptide Mass

Noroviruses (NVs) are one of the leading causes of epidemic gastroenteritis around the world. Water treatment technologies using membrane filtration for virus removal are becoming increasingly important. However, experiments to test removal of NVs from water have been hampered because NVs do not grow in cell culture or in small-animal models and therefore cannot be easily artificially propagated. Expression of the NV genome in a baculovirus-silkworm expression system has produced recombinant NV virus-like particles (rNV-VLPs) that are morphologically and antigenically similar to native NV. Here, we characterized these rNV-VLPs and evaluated their potential use in assessing NV removal. Electron microscopic analysis and peptide mass fingerprinting showed that the rNV-VLPs were morphologically identical to native NV. In addition, surface charge and particle size distribution, which are important factors for explaining virus particle behavior during membrane filtration, were successfully evaluated by using rNV-VLPs. The rNV-VLPs were easy to quantify with a commercially available enzyme-linked immunosorbent assay kit, they remained stable for several days at 4 °C after dilution in river water, and they were easy to concentrate with the ultrafiltration entrapment method used. Thus, rNV-VLPs can be used to facilitate our understanding of the behavior of NVs during membrane filtration processes.

scholarly journals Development of Rapid Immunochromatographic Test with Recombinant NcSAG1 for Detection of Antibodies to Neospora caninum in Cattle

2005 ◽  
Vol 12 (7) ◽  
pp. 885-887 ◽  
Author(s):  
Min Liao ◽  
Shoufa Zhang ◽  
Xuenan Xuan ◽  
Guohong Zhang ◽  
Xiaohong Huang ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Rapid Detection ◽  
Reliable Method ◽  
Neospora Caninum ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunochromatographic Test ◽  
Serum Samples ◽  
Immunofluorescent Antibody Test ◽  
Good Agreement

ABSTRACT An immunochromatographic test (ICT) with recombinant surface antigen 1 of Neospora caninum (NcSAG1) was developed for the rapid detection of antibodies to N. caninum in cattle. The ICT was used to clearly discriminate between immunofluorescent-antibody test (IFAT)-positive bovine sera and IFAT-negative bovine sera. Serum samples collected from cattle in Yanbian, China, were examined by the ICT. Of the 96 serum samples, 23 (24.0%) were positive by the ICT, and 19 (19.8%) samples were positive by a previously developed enzyme-linked immunosorbent assay (ELISA). Eighteen of 19 ELISA-positive samples were positive according to the ICT. A good agreement was found between the results of the ICT and the ELISA. The results presented here suggest that the ICT with recombinant truncated NcSAG1 fused to glutathione S-transferase is a useful and reliable method for the detection of antibodies to N. caninum in cattle.

scholarly journals Evaluation of the performance of three serological tests for diagnosis of Leishmania infantum infection in dogs using latent class analysis

2020 ◽  
Vol 29 (4) ◽  
Author(s):  
Asier Basurco ◽  
Alda Natale ◽  
Katia Capello ◽  
Antonio Fernández ◽  
María Teresa Verde ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Leishmania Infantum ◽  
Latent Class ◽  
Rapid Test ◽  
Antibody Test ◽  
Latent Class Models ◽  
Canine Leishmaniasis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests

Abstract Canine leishmaniasis (CanL) is a disease caused by Leishmania infantum. Serological methods are the most common diagnostic techniques used for the diagnosis of the CanL. The objective of our study was to estimate the sensitivity and specificity of one in-house ELISA kit (ELISA UNIZAR) and three commercially available serological tests (MEGACOR Diagnostik GmbH) including an immunochromatographic rapid test (FASTest LEISH®), an immunofluorescent antibody test (MegaFLUO LEISH®) and an enzyme-linked immunosorbent assay (MegaELISA LEISH®), using latent class models in a Bayesian analysis. Two hundred fifteen serum samples were included. The highest sensitivity was achieved for FASTest LEISH® (99.38%), ELISA UNIZAR (99.37%), MegaFLUO LEISH® (99.36%) followed by MegaELISA LEISH® (98.49%). The best specificity was obtained by FASTest LEISH® (98.43%), followed by ELISA UNIZAR (97.50%), whilst MegaFLUO LEISH® and MegaELISA LEISH® obtained the lower specificity (91.94% and 91.93%, respectively). The results of present study indicate that the immunochromatographic rapid test evaluated FASTest LEISH® show similar levels of sensitivity and specificity to the quantitative commercial tests. Among quantitative serological tests, sensitivity and specificity were similar considering ELISA or IFAT techniques.

scholarly journals AVIAN INFLUENZA ANTIBODY RESPONSE IN PIGLETS THATAREGIVEN ESCHERICHIA COLI–AVIAN INFLUENZA VACCINES

2021 ◽  
Vol 10 (1) ◽  
pp. 61-70
Author(s):  
Audrey Febiannya Putri Bhaskara ◽  
I Gusti Ngurah Kade Mahardika ◽  
I Nyoman Suartha
Keyword(s):  
Escherichia Coli ◽  
Avian Influenza ◽  
Optical Density ◽  
Antibody Test ◽  
Influenza Vaccines ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Antibody ◽  
E Coli ◽  
Test Kit ◽  
Virus Influenza

Babi berperan penting dalam ekologi virus influenza, karena babi dapat berperan sebagai wahana untuk reasorsi virus influenza dari unggas dan mamalia. Vaksinasi dengan antigen influenza universal, yaitu nukleoprotein, dapat menurunkan peluang babi dalam memunculkan virus influenza baru. Penelitian ini bertujuan untuk mengentahui respons antibodi dari vaksinasi dengan nukleoprotein rekombinan - Escherichia coli. Sebanyak 12 anak babi landrace dari tiga induk yang berbeda dipilih secara acak. Enam ekor divaksinasi dengan vaksin nucleoprotein-E. coli pada umur tujuh hari dan diulang pada umur 21 hari. Enam ekor tidak divaksinasi. Serum diambil pada umur 35 hari. Nilai optical density (OD) antibodi terhadap nukleoprotein diuji dengan teknik Enzyme-Linked Immunosorbent Assay (ELISA) dengan menggunakan Kit ELISA komersial, Avian Influenza Virus Antibody Test Kit. Hasil penelitian menunjukkan bahwa nilai Optical Density rata-rata babi yang divaksinasi (0,367) secara statistika nyata lebih tinggi dibandingkan dengan yang tidak divaksinasi (0,054). Vaksin rekombinan nucleoprotein-E. coli yang dicobakan mampu meningkatan antibodi terhadap virus avian influenza pada anak babi.

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