scholarly journals Evaluation of a commercial Erns-capture ELISA for detection of BVDV in routine diagnostic cattle serum samples

2007 ◽  
Vol 49 (1) ◽  
Author(s):  
Jaruwan Kampa ◽  
Karl Ståhl ◽  
Lena HM Renström ◽  
Stefan Alenius
Keyword(s):  
Serum Samples ◽  
Capture Elisa ◽  
Cattle Serum

scholarly journals Development of Recombinant Nucleoprotein-Based Diagnostic Systems for Lassa Fever

2007 ◽  
Vol 14 (9) ◽  
pp. 1182-1189 ◽  
Author(s):  
Masayuki Saijo ◽  
Marie-Claude Georges-Courbot ◽  
Philippe Marianneau ◽  
Victor Romanowski ◽  
Shuetsu Fukushi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Hela Cells ◽  
Recombinant Baculovirus ◽  
Lassa Fever ◽  
Rabbit Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lassa Virus ◽  
Diagnostic Systems ◽  
Serum Samples ◽  
Capture Elisa

ABSTRACT Diagnostic systems for Lassa fever (LF), a viral hemorrhagic fever caused by Lassa virus (LASV), such as enzyme immunoassays for the detection of LASV antibodies and LASV antigens, were developed using the recombinant nucleoprotein (rNP) of LASV (LASV-rNP). The LASV-rNP was expressed in a recombinant baculovirus system. LASV-rNP was used as an antigen in the detection of LASV-antibodies and as an immunogen for the production of monoclonal antibodies. The LASV-rNP was also expressed in HeLa cells by transfection with the expression vector encoding cDNA of the LASV-NP gene. An immunoglobulin G enzyme-linked immunosorbent assay (ELISA) using LASV-rNP and an indirect immunofluorescence assay using LASV-rNP-expressing HeLa cells were confirmed to have high sensitivity and specificity in the detection of LASV-antibodies. A novel monoclonal antibody to LASV-rNP, monoclonal antibody 4A5, was established. A sandwich antigen capture (Ag-capture) ELISA using the monoclonal antibody and an anti-LASV-rNP rabbit serum as capture and detection antibodies, respectively, was then developed. Authentic LASV nucleoprotein in serum samples collected from hamsters experimentally infected with LASV was detected by the Ag-capture ELISA. The Ag-capture ELISA specifically detected LASV-rNP but not the rNPs of lymphocytic choriomeningitis virus or Junin virus. The sensitivity of the Ag-capture ELISA in detecting LASV antigens was comparable to that of reverse transcription-PCR in detecting LASV RNA. These LASV rNP-based diagnostics were confirmed to be useful in the diagnosis of LF even in institutes without a high containment laboratory, since the antigens can be prepared without manipulation of the infectious viruses.

scholarly journals Evaluation of an Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Orientia tsutsugamushi Infection

2003 ◽  
Vol 10 (3) ◽  
pp. 394-398 ◽  
Author(s):  
Won-Jong Jang ◽  
Myung-Suk Huh ◽  
Kyung-Hee Park ◽  
Myung-Sik Choi ◽  
Ik-Sang Kim
Keyword(s):  
Immunosorbent Assay ◽  
Scrub Typhus ◽  
Microtiter Plate ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Orientia Tsutsugamushi ◽  
Serum Samples ◽  
Capture Elisa ◽  
Igm Antibodies ◽  
Immunodominant Antigen

ABSTRACT To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We developed an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) for diagnosis of recent Orientia tsutsugamushi infections in humans. The 56-kDa major outer membrane protein of O. tsutsugamushi is well known as the most immunodominant antigen in scrub typhus. The test is based on the use of the biotinylated recombinant 56-kDa protein of O. tsutsugamushi Boryong, Bor56, which was expressed as a fusion protein with a maltose-binding protein in Escherichia coli. In the test, the serum IgM antibodies were captured by anti-human IgM antibodies coated onto a microtiter plate. The captured IgM antibodies were revealed through sequential addition of biotinylated Bor56 antigen and peroxidase-conjugated streptavidin to the plate. The IgM capture ELISA was compared with the immunofluorescence antibody assay (IFA) by testing 176 serum samples from patients with diagnosed cases of rickettsial disease and patients with other acute febrile diseases. Of the 81 IgG IFA-positive samples, 78 tested positive (sensitivity, 96.3%) and all 31 IgM IFA-positive samples tested positive (sensitivity, 100%) by the IgM capture ELISA. The specificity of the IgM capture ELISA was 99%, and 1 of the 95 IFA-negative samples was positive in the assay. These results strongly suggest that IgM capture ELISA using the recombinant Bor56 antigen is a reliable and detailed method for the detection of early O. tsutsugamushi infection.

scholarly journals A μ-capture ELISA for detecting Mycoplasma pneumoniae IgM: comparison with indirect immunofluorescence and indirect ELISA

1985 ◽  
Vol 94 (2) ◽  
pp. 217-227 ◽  
Author(s):  
T. G. Wreghitt ◽  
Margaret Sillis
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Indirect Immunofluorescence ◽  
Indirect Elisa ◽  
Test Results ◽  
Serum Samples ◽  
Gradual Decline ◽  
Capture Elisa ◽  
Diagnostic Use ◽  
Antibody Levels ◽  
Indirect Immunofluorescent

SUMMARYA μ-capture ELISA was developed for detecting Mycoplasma pneumoniae specific IgM, and compared with an indirect immunofluorescent antibody (IFA) technique and an indirect ELISA. μ-capture ELISA and IFA compared well and were found to be the most sensitive assays. The IFA test can be completed in 2 h whilst the results of the μ-capture ELISA can be available in 24 h. Both tests are amenable to routine diagnostic use and have similar sensitivity. Indirect ELISA was found to be less sensitive and less specific, giving high assay values with several sera having undetectable M. pneumoniae CF antibody or CF antibody in low titre. Serum samples obtained from 11 patients at various times after M. pneumoniae infection showed maximum antibody levels within the first month by all assays, with a gradual fall in amount of IgM with time when assayed by μ-capture ELISA, a more gradual decline by IFA and hardly any decline with indirect ELISA. It was concluded that the indirect ELISA is unsuitable for the investigation of possible M. pneumoniae infection because the sustained high assay values with serum samples taken many months after infection, make interpretation of the test results very difficult.

scholarly journals Performance of the Dot-blot test method for detecting antibodies to Sarcocystis spp. in cattle

2020 ◽  
Vol 40 (5) ◽  
pp. 385-388
Author(s):  
Maiara S.T. Ferreira ◽  
Fagner D. Fernandes ◽  
Marta E.M. Alves ◽  
Patricia Bräunig ◽  
Luis A. Sangioni ◽  
...  
Keyword(s):  
Neospora Caninum ◽  
Standard Technique ◽  
Good Alternative ◽  
Test Method ◽  
Dot Blot ◽  
Serum Samples ◽  
Sarcocystis Spp ◽  
Cattle Serum ◽  
Antibody Tests ◽  
Gold Standard Technique

ABSTRACT: Serological techniques can detect antibodies against Sarcocystis spp., Neospora caninum and Toxoplasma gondii antigens in single or mixed infections. Immunofluorescent antibody tests (IFAT) is considered the gold standard technique for Sarcocystosis diagnostic in cattle serum and a positive IFAT result reflects Sarcocystis spp. infection. Therefore, the aims of the present study were to compare IFAT and Dot-blot for sarcocystosis diagnostic in experimentally infected mice and to investigate serological cross-reactions with N. caninum and T. gondii in these methods. Mice (Mus musculus) were inoculated intraperitoneally with bradizoites of Sarcocystis spp. or tachyzoites of N. caninum or T. gondii. Serum samples were obtained and analyzed by IFAT and Dot-blot for the three protozoa. Serum from N. caninum and T. gondii experimentally infected mice were tested by IFAT and reacted only to N. caninum or T. gondii antigens, respectively. Specific antibodies against Sarcocystis spp. were present in all animals experimentally infected with this protozoan, with IFAT titers from 10 to 800. Serum samples from mice experimentally infected with Sarcocystis spp., N. caninum and T. gondii and tested by Dot-blot demonstrated no cross reaction between protozoa. A Dot-blot using Sarcocystis spp. antigen appears to be a good alternative to IFAT in the serological diagnosis of Sarcocystosis.

scholarly journals Detection of Vaccinia Virus in Dairy Cattle Serum Samples from 2009, Uruguay

2016 ◽  
Vol 22 (12) ◽  
pp. 2174-2177 ◽  
Author(s):  
Ana Paula Moreira Franco-Luiz ◽  
Danilo Bretas Oliveira ◽  
Alexandre Fagundes Pereira ◽  
Mirela Cristina Soares Gasparini ◽  
Cláudio Antônio Bonjardim ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Vaccinia Virus ◽  
Serum Samples ◽  
Cattle Serum

scholarly journals Patterns of Rift Valley fever activity in Zambia

1992 ◽  
Vol 108 (1) ◽  
pp. 185-191 ◽  
Author(s):  
F. G. Davies ◽  
E. Kilelu ◽  
K. J. Linthicum ◽  
R. G. Pegram
Keyword(s):  
Remote Sensing ◽  
Satellite Imagery ◽  
Rift Valley ◽  
Rift Valley Fever ◽  
Rift Valley Fever Virus ◽  
Serum Samples ◽  
Valley Fever ◽  
Low Level ◽  
Virus Activity ◽  
Cattle Serum

SUMMARYSummary An hypothesis that there was an annual emergence of Rift Valley fever virus in Zambia, during or after the seasonal rains, was examined with the aid of sentinel cattle. Serum samples taken during 1974 and 1978 showed evidence of epizootic Rift Valley fever in Zambia, with more than 80% positive. A sentinel herd exposed from 1982 to 1986 showed that some Rift Valley fever occurred each year. This was usually at a low level, with 3–8% of the susceptible cattle seroconverting. In 1985–6 more than 20% of the animals seroconverted, and this greater activity was associated with vegetational changes - which could be detected by remote-sensing satellite imagery-which have also been associated with greater virus activity in Kenya.

scholarly journals DISTRIBUTION OF BOVINE HERPESVIRUS 1 IN CATTLE POPULATION AND BULLS FROM CENTERS FOR ARTIFICIAL INSEMINATION AND PRIVATE FARMS IN BULGARIA

2013 ◽  
Vol 6 (1) ◽  
pp. 3-17
Author(s):  
Raiko Peshev ◽  
Lilly Christova
Keyword(s):  
Antibody Detection ◽  
Gene Detection ◽  
Serum Samples ◽  
Bovine Herpesvirus 1 ◽  
Serological Examination ◽  
Elisa Testing ◽  
Vaginal Swabs ◽  
Cattle Serum ◽  
Herpesvirus 1

The aim of the study is determination of the BHV 1 incidence among the bovine population in Bulgaria by using various ELISA tests for antibody detection and diff erent methods for proving viral antigen. Commercial diagnostic ELISA kits were used for screening as well as for confi rmation of the antibodies. Serological examination encompassed 2973 serum samples from bovine population in Bulgaria (cattle, calves and bulls) originating from 21 country regions. Total 408 cattle and 150 bulls’ samples originating from artifi cial insemination centers (AIC), commune and private farms from 21 country regions were subjected to virology testing. Identifi cation of isolated viruses was performed using conventional and nested PCRs for BHV 1 gB gene detection. Th e percentage of positively reacting cattle sera were signifi cantly (p < 0.001) higher (38.3%) then that of bulls (29.3%) after ELISA testing of 2240 cattle serum samples and 733 bulls’ sera. Seven strains with BHV 1 characteristics were isolated from Dobrich, Lovech – 2 strains, Plovdiv, Targovishte, Pazardjik and Svishtov regions after cultivation 408 samples from cattle and calves (buff y coats, nasal, eyes, vaginal swabs, tissues and organs) on cell cultures. Th ree BHV 1 strains were isolated from Sliven, Shumen and Blagoevgrad regions aft er examining 150 bulls’ samples (nasal, eyes, preputial, semen samples and buff y coats). Confi rmation of virus isolation was accomplished using PCR gB gene based primers (amplified 478 bp) Th e amplicons from 3 bulls isolated strains with the same size and location as the referent Oxford strain were observed. After performance of nested PCR for gB gen, products with size of 385 bp were obtained in all samples as well as in the referent Oxford strain.

Seroprevalence of infectious bovine rhinotracheitis in organized dairy farms of India

2015 ◽  
Author(s):  
S. S. Patil ◽  
A. Prajapati ◽  
P. Krishnamoorthy ◽  
G. S. Desai ◽  
G. B.M. Reddy ◽  
...  
Keyword(s):  
Medical History ◽  
Central Region ◽  
Age Groups ◽  
Dairy Farms ◽  
Northern Region ◽  
Infectious Bovine Rhinotracheitis ◽  
Serum Samples ◽  
History Of ◽  
Cattle Serum

In this study, a systematic sero-surveillance of IBR was undertaken from 11 dairy farms located in 4 different regions of India. A total of 1000 cattle serum samples were tested for the presence of antibodies against IBR using Avidin Biotin ELISA. The results revealed that IBR antibodies were widely prevalent in all regions of the country ranging from 36.5% in Central region to 84.5% in Northern region with an overall prevalence of 61.6%. The prevalence of IBR antibodies was different between various age groups being 22.3%, 62.1%, 59.3%, 76.1% and 66.78 % in the age groups less than 1 year, 1-2 years, 2-3 years, 3-4 years and more than 4 years old respectively. Based upon the medical history of the herd, it was found that 83% abortion cases, 76% metritis cases, 83% repeat breeding cases and 65% retention of placenta cases were seropositive for IBR.

scholarly journals PREVALENCE OF COXIELLA BURNETII INFECTION IN CATTLE, BLACK BENGAL GOATS AND TICKS IN BANGLADESH

2016 ◽  
Vol 14 (1) ◽  
pp. 65-68 ◽  
Author(s):  
A. Chakrabartty ◽  
P. K. Bhattacharjee ◽  
R. R. Sarker ◽  
A. K. M. A. Rahman ◽  
K. Henning ◽  
...  
Keyword(s):  
Real Time ◽  
Coxiella Burnetii ◽  
Real Time Pcr ◽  
Q Fever ◽  
Elisa Test ◽  
Quantitative Real Time Pcr ◽  
Serum Samples ◽  
Test Kit ◽  
History Of ◽  
Cattle Serum

The objectives of this study were to determine the prevalence of Coxiella burnetii infection in domestic ruminants and to detect Coxiella burnetii DNA from ticks and serum samples. A total of 24 ticks, 91 goats and 81 cattle serum samples with the history of abortion and reproductive disorders were collected from the different areas in Bangladesh. The serum samples were tested by CHEKIT Q-Fever Antibody ELISA Test Kit and Coxiella burnetii DNA was detected by multiplex quantitative real- time PCR. The overall prevalence was 7.6% and 6.1% in goats and cattle, respectively. However, none of seropositive samples and tick samples was positive in quantitative real-time PCR.

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