scholarly journals Comparison of enzyme-linked immunosorbent assay and RT-PCR for the detection of porcine epidemic diarrhoea virus

2010 ◽  
Vol 88 (1) ◽  
pp. 166-168 ◽  
Author(s):  
Enrica Sozzi ◽  
Andrea Luppi ◽  
Davide Lelli ◽  
Ana Moreno Martin ◽  
Elena Canelli ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Porcine Epidemic Diarrhoea Virus ◽  
Diarrhoea Virus

scholarly journals Influence of category, herd size, grazing and management on epidemiology of bovine viral diarrhoea in dairy herds

2013 ◽  
Vol 82 (2) ◽  
pp. 125-130 ◽  
Author(s):  
Tomislav Bedeković ◽  
Nina Lemo ◽  
Ljubo Barbić ◽  
Željko Cvetnić ◽  
Ivana Lojkić ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Production Management ◽  
Herd Size ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dairy Herds ◽  
Bovine Viral Diarrhoea ◽  
Tissue Samples ◽  
Persistently Infected ◽  
Diarrhoea Virus ◽  
High Prevalence

The aim of this study was to estimate the influence of category, herd size, common grazing and management as risk factors in maintaining bovine viral diarrhoea infection in dairy herds. A total of 987 sera samples obtained from 202 heifers, 653 cows and 132 calves from 103 herds in Croatia were examined by enzyme-linked immunosorbent assay. In order to establish the prevalence of persistently infected cattle, 35 herds were selected. Ear notch tissue samples from all animals in selected herds (n = 2284) were collected and analyzed by antigen enzyme-linked immunosorbent assay. The true prevalence of specific antibodies was 61.61% and the estimated prevalence of exposure to bovine viral diarrhoea virus at the herd level was 100%. The prevalence of persistently infected animals was 0.53% and the prevalence of persistently infected herds was 20%. The antibodies prevalence was higher in cows, in herds that use common pasture and in larger herds (P < 0.001). The prevalence of persistently infected animals was not connected with the herd size but production management on big farms contributed to maintaining the virus. The obtained results suggest that production management was an important risk factor in bovine viral diarrohea epidemiology. High prevalence of antibodies and high prevalence of persistently infected herds requires implementation of control and eradication programs at a national or even regional level. The presented data complete the BVD epidemiological investigations from this part of Europe.

Evaluation of a Sensitive Reverse Transcription PCR–Enzyme-Linked Immunosorbent Assay for Detection of Hepatitis A Virus in Oysters (Saccostrea glomerata) on the East Coast of the Gulf of Thailand

2014 ◽  
Vol 77 (5) ◽  
pp. 859-863 ◽  
Author(s):  
URAIWAN INTAMASO ◽  
SITTHISAK KETKHUNTHOD
Keyword(s):  
Immunosorbent Assay ◽  
Reverse Transcription ◽  
Hepatitis A ◽  
East Coast ◽  
Hepatitis A Virus ◽  
Detection Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Gulf Of Thailand ◽  
Rt Pcr ◽  
The Gulf Of Thailand

Hepatitis A virus (HAV) contamination in food can lead to major health problems. We developed a combination reverse transcription (RT) PCR method plus enzyme-linked immunosorbent assay (ELISA) to detect HAV in fresh oysters harvested along the east coast of the Gulf of Thailand. Viral nucleic acid was extracted via the glycine–arginine–polyethylene glycol method followed by RT-PCR amplification with specifically designed primers against HAV and an ELISA to detect the digoxigenin-labeled RT-PCR products. The ELISA in concert with the RT-PCR protocol further increased the detection sensitivity by 100-fold for the HAV genome and 10-fold in artificially contaminated oysters. The overall sensitivity of the RT-PCR in combination with the ELISA was 31.88 pg and 16 PFU/g, respectively. The ELISA increases the specificity of the RT-PCR assay for detecting naturally occurring HAV in oysters. This combined RT-PCR-ELISA approach is a practical and sensitive method for HAV detection and can be utilized in routine screening for HAV in shellfish.

scholarly journals Development of rapid immunochromatographic strip test for the detection of porcine epidemic diarrhoea virus

2017 ◽  
Vol 181 (22) ◽  
pp. 596-601 ◽  
Author(s):  
Kwang-Soo Lyoo ◽  
Minjoo Yeom ◽  
Jungho Kim ◽  
Donghyuk Kim ◽  
Gunwoo Ha ◽  
...  
Keyword(s):  
Real Time ◽  
Rapid Test ◽  
Epidemiological Surveillance ◽  
Watery Diarrhoea ◽  
Rt Pcr ◽  
Immunochromatographic Strip ◽  
Test Kit ◽  
Porcine Epidemic Diarrhoea Virus ◽  
Diarrhoea Virus ◽  
Strip Test

Porcine epidemic diarrhoea virus (PEDV) causes acute and severe watery diarrhoea and dehydration, as well as 50–100 per cent mortality in piglets. For the PEDV diagnosis, a rapid test kit that is specific and sensitive to PEDV is critical to monitor this disease at pig farms. The present study aimed to develop an immunochromatographic assay (ICA) strip test for detecting PEDV in faecal swabs. The newly developed diagnostic test showed a detection limit of 104.0TCID50/ml of PEDV. Using faecal swab samples, the relative sensitivity and specificity of the ICA kit were 95.0 per cent and 98.6 per cent, respectively, compared with those of real-time RT-PCR. In samples from piglets experimentally infected with PEDV, the results showed 100 per cent agreement with those found by real-time RT-PCR. Our developed test strip will be useful for rapid diagnosis and can be used for epidemiological surveillance of PEDV infection.

scholarly journals Development and Validation of an Enzyme-Linked Immunosorbent Assay for Human Metapneumovirus Serology Based on a Recombinant Viral Protein

2005 ◽  
Vol 12 (2) ◽  
pp. 249-253 ◽  
Author(s):  
Marie-Ève Hamelin ◽  
Guy Boivin
Keyword(s):  
Immunosorbent Assay ◽  
Serological Test ◽  
Human Metapneumovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chronic Obstructive ◽  
Rt Pcr ◽  
Recent Infection ◽  
Hmpv Infection ◽  
Group A ◽  
Syncytial Virus

ABSTRACT The human metapneumovirus (hMPV) is a newly reported respiratory virus belonging to the Paramyxoviridae family that has been associated with bronchiolitis and pneumonia in young children. We developed a simple enzyme-linked immunosorbent assay (ELISA) for hMPV serological testing using the nucleoprotein (N) from group A or B (N-A or N-B) as the antigen, and we evaluated it in both children and adults. The N proteins were first used in a Western immunoblot assay to identify hMPV-negative sera, which were then used to determine the cutoff value of the ELISA test. Subsequent evaluation of the ELISA-N test revealed that the mean reciprocal antibody titer of 20 randomly selected seropositive children was 143, compared to 69 for 20 seropositive adults. In a prospective evaluation of 71 adults with acute exacerbations of chronic obstructive pulmonary disease, 58 (81.6%) had prior hMPV antibodies and 3 (4.2%) had evidence of recent hMPV infection. In testing paired sera from adults (n = 4) with recent hMPV group A infection confirmed by reverse transcriptase PCR (RT-PCR), ELISAs using the N-A or N-B proteins were able to detect hMPV seroconversion. Moreover, testing of paired sera from three adults with a recent infection by the human respiratory syncytial virus confirmed by RT-PCR and serology did not reveal any increase in hMPV antibodies over time. The ELISA-N is a simple, objective, and specific serological test useful for detecting anti-hMPV antibodies following group A or B viral infections, which should permit a better understanding of the epidemiology of this virus.

scholarly journals Development and application of a TaqMan-MGB real-time RT-PCR assay for the detection of porcine epidemic diarrhoea virus strains in China

2016 ◽  
Vol 60 (2) ◽  
pp. 127-133 ◽  
Author(s):  
Yi-Xuan Hou ◽  
Chun Xie ◽  
Kang Wang ◽  
Yu-Ting Zhao ◽  
Yang-Yang Xie ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
N Gene ◽  
Quantitative Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Conserved Sequence ◽  
Porcine Epidemic Diarrhoea Virus ◽  
Diarrhoea Virus ◽  
Quantitative Pcr Assay

AbstractIntroduction:A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed.Material and Methods:Based on the conserved sequence of the PEDV nucleocapsid (N) gene, a primer pair and probe were designed to establish a TaqMan-MGB real-time RT-PCR assay for quantitative detection of the virus. The sequence was cloned into the pMD18-T vector and a series of diluted recombinant plasmids were used to generate a standard curve with an R2 value of 0.999.Results:The developed quantitative PCR assay detected viral titres as low as 0.1 TCID50with high specificity and no cross-reaction with other porcine viruses (PoRV, TGEV, PRRSV, or CSFV). The intra-batch and inter-batch coefficients of variation were both less than 1%, which indicated good reproducibility. Thirty clinical diarrhoea samples obtained from pigs in Shanghai and Fujian were analysed using this quantitative PCR assay. Out of these samples, 93.3% were found to be PEDV positive.Conclusion:This approach is suitable for clinical sample identification and pathogenesis studies.

scholarly journals Allergene Lipide und ihre Bedeutung für die Allergiediagnostik

2017 ◽  
Vol 5 (4) ◽  
pp. 207-208
Author(s):  
Claudia Traidl-Hoffmann
Keyword(s):  
Immunosorbent Assay ◽  
Antigen Presenting Cells ◽  
Buffy Coat ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dendritische Zellen ◽  
Rt Pcr ◽  
Tnf Α ◽  
Allergische Sensibilisierung ◽  
Olea Europea ◽  
Antigen Presenting

Hintergrund: Die allergische Sensibilisierung wird möglicherweise durch Lipide beeinflusst, die in den Allergenen enthalten sind. Dies lässt sich an der Reaktion natürlicher Killer-T-Zellen (NKT-Zellen) mit Antigen-präsentierenden Zellen (antigen-presenting cells, APC) erkennen. Das Ziel dieser Studie war die Untersuchung der Auswirkungen von Olivenpollenlipiden auf humane APC einschließlich Monozyten sowie aus Monozyten hervorgegangene Makrophagen (Mϕ) und dendritische Zellen (DZ). Methoden: Aus Pollenstaub des Olivenbaums (Olea europea) wurden Lipide extrahiert. Invariante NKT-Zellen (iNKT-Zellen), Monozyten, Mϕ und DZ wurden aus dem Buffy Coat von Blutspenden gesunder Spender gewonnen, und der Zell-Phänotyp wurde mit Hilfe der Durchflusszytometrie bestimmt. Die Beurteilung der Zytotoxizität von iNKT-Zellen erfolgte durch ein Laktatdehydrogenase-Assay. Die Genexpression von CD1A und CD1D wurde mittels RT-PCR bestimmt. Die Produktion der Zytokine IL-6, IL-10, IL-12 und TNF-α durch Monozyten, Mϕ, und DZ wurde unter Verwendung des Enzyme-linked Immunosorbent Assay (ELISA) gemessen. Ergebnisse: Unsere Ergebnisse belegen, dass Monozyten und Mϕ nach Behandlung mit Olivenpollenlipiden in hohem Maße iNKT-Zellen aktivieren. Wir beobachteten mehrere Veränderungen im Phänotyp der APC nach Exposition gegenüber den Pollenlipiden. Sowohl bei den Mϕ als auch bei den Monozyten stieg nach der Behandlung mit den Olivenpollenlipiden die CD1D-Genexpression, während eine Hochregulierung des CD1d-Zelloberflächenproteins nur bei den Mϕ eintrat. Auch im Humanserum differenzierte DZ steigerten ihre CD1d-Oberflächenexpression nach Exposition gegenüber Olivenpollenlipiden. Außerdem vermochten Olivenpollenlipide die IL-6-Produktion zu stimulieren, die Produktion von Lipopolysaccharid-induziertem IL-10 durch Mϕ hingegen wurde herunterreguliert. Schlussfolgerungen: Olivenpollenlipide bewirken Veränderungen im Phänotyp von Monozyten, Mϕ und DZ, die zur Aktivierung von NKT-Zellen führen, was wiederum potenziell die allergische Immunreaktion beeinflussen kann.

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