Development and validation of a monoclonal antibody blocking elisa for the detection of antibodies against both equine herpesvirus type 1 (EHV1) and equine herpesvirus type 4 (EHV4)

Maanen, C. van; Boer-Luijtze, E de; Terpstra, C.

doi:10.1016/s0378-1135(99)00162-5

Cited by 16 publications

(7 citation statements)

References 19 publications

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“…The necessity to examine undiluted as well as diluted samples in a similar blocking assay was recently described for horse sera containing antibodies to equine arteritis virus (Cho et al., 2000). Using flow cytometry or immunofluoresence microscopy for such an assay might sound inconvenient when compared with an ELISA format as has been described for the serological discrimination of a number of pathogens including viruses, bacteria, parasites, fungi or antitoxins (Hall et al., 1995; Kolbe and Clough, 1999; Garringer et al., 2000; van Maanen et al., 2000; Nugent et al., 2000; Saliki and Lehenbauer, 2001). This is especially true because ELISA readers are considered to be part of any diagnostic laboratory in the world where the former two equipments are not that ubiquitously available.…”

Section: Discussionmentioning

confidence: 99%

Detection of Antibodies to Alphaviruses and Discrimination between Antibodies to Eastern and Western Equine Encephalitis Viruses in Rabbit Sera using a Recombinant Antigen and Virus‐specific Monoclonal Antibodies

Pässler

Pfeffer²

2003

Journal of Veterinary Medicine, Series B

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Three arthropod-borne alphaviruses, western equine encephalitis viruses (WEEV), eastern equine encephalitis viruses (EEEV) and Venezuelan equine encephalitis viruses are the aetiological agents of a sometimes severe encephalomyelitis in equines and humans in the New World. With regard to the different ecology and epidemiology of these viruses, a method applied in serological screening should be able to distinguish between them as well as other related members of the genus Alphavirus in the American continent. However, this has been hampered in the past by (a) the close antigenic relationship between alphaviruses in traditional serological assays, especially in the routinely used haemagglutination-inhibition, and (b) the need of biosafety level 3 facilities to grow the viral antigens. An epitope blocking assay using an EEEV glycoprotein E1-expressing recombinant Sindbis virus and virus-specific monoclonal antibodies (mAbs) binding to the E1 of EEEV (strain NJ/60) and the E1 of Sindbis virus was established using automated flow cytometry. The test was evaluated using sera of infected and vaccinated rabbits. A cut-off value of 30% inhibition for antigenic complex-specific seroconversion was found to be sufficient for the detection of the respective infection. By using three different mAbs in parallel, we were able to detect alphavirus genus-, EEEV- and WEEV-complex-specific serum antibodies. As this test is based on the inhibition of binding of virus-specific mAbs, sera of every origin other than mouse can be tested. Thus, this assay may prove useful in the serological screening of a variety of animal species during an outbreak investigation.

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Section: Discussionmentioning

confidence: 99%

Detection of Antibodies to Alphaviruses and Discrimination between Antibodies to Eastern and Western Equine Encephalitis Viruses in Rabbit Sera using a Recombinant Antigen and Virus‐specific Monoclonal Antibodies

Pässler

Pfeffer²

2003

Journal of Veterinary Medicine, Series B

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“…To improve serological diagnosis, we developed a rapid, reproducible and sensitive monoclonal antibody blocking ELISA for the detection of antibodies against EHV1 and EHV4. This ELISA is easy to perform, economic in use of reagents and does not require the use of both EHV1 and EHV4 strains for serological diagnosis of an EHV1 or EHV4 infection (171). Moreover, this ELISA appeared to be more sensitive than a homologous VN test to detect responses after EHV1/4 infections early in life and after experimental infections (171).…”

Section: Additional Comments On Serodiagnosis Of Ehv1 and Ehv4 Infectmentioning

confidence: 96%

Equine herpesvirus 1 and 4 infections: An update

Maanen

2002

Veterinary Quarterly

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Summary Equine herpesvirus 1 (EHV1) and equine herpesvirus 4 (EHV4) are important ubiquitous equine pathogens, causing much damage to the viral horse industry. EHV1 strains are associated with respiratory disease, abortion, and paresis/paralysis, whereas EHV4 strains are predominantly associated with respiratory disease. In the past decades much research effort has been put into improving knowledge about these viruses. In this paper the current state of knowledge of these viruses and the most important aspects of these virus infections, e.g. epidemiology, clinical aspects, pathogenesis and pathology, immunity, diagnosis, preventive management and management in the course of an outbreak and vaccination, is reviewed. Because we performed some research ourselves in the areas of diagnosis, epidemiology and vaccinology these aspects are reviewed in more depth than the other aspects. Still many questions have remained and new questions have risen. Consequently, research priorities should be made in an attempt to answer these questions. Therefore, this review ends with some personal recommendations for important priorities for future research.

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“…Blocking ELISA, a specific method for antibody detection, has been used widely to diagnose human diseases [18], monitor animal infectious diseases [19], [20], [21], [22], [23], and detect viral antibodies in clinics or laboratories [24], [25], [26], [27], [28], [29]. The distinct advantages of blocking ELISA include high-volume sample testing, applicability across multiple species, and greater objectivity than some traditional techniques.…”

Section: Discussionmentioning

confidence: 99%

Development of a Blocking ELISA for Detection of Serum Neutralizing Antibodies against Newly Emerged Duck Tembusu Virus

Teng

et al. 2012

PLoS ONE

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BackgroundSince April 2010, domesticated ducks in China have been suffering from an emerging infectious disease characterized by retarded growth, high fever, loss of appetite, decline in egg production, and death. The causative agent was identified as a duck Tembusu virus (DTMUV), a member of the Ntaya virus (NTAV) group within the genus Flavivirus, family Flaviviridae. DTMUV is highly contagious and spreads rapidly in many species of ducks. More than 10 million shelducks have been infected and approximately 1 million died in 2010. The disease remains a constant threat to the duck industry; however, it is not known whether DTMUV can infect humans or other mammalians, despite the fact that the virus has spread widely in southeast China, one of the most densely populated areas in the world. The lack of reliable methods to detect the serum antibodies against DTMUV has limited our ability to conduct epidemiological investigations in various natural hosts and to evaluate the efficiency of vaccines to DTMUV.Methodology/Principal FindingsA neutralizing monoclonal antibody (mAb) 1F5 binding specifically to the E protein was developed. Based on the mAb, a blocking enzyme-linked immunosorbent assay (ELISA) was developed for the detection of neutralizing antibodies against DTMUV. The average value of percent inhibition (PI) of 350 duck serum samples obtained from DTMUV-free farms was 1.0% ±5.8% (mean ± SD). The selected cut-off PI values for negative and positive sera were 12.6% (mean +2SD) and 18.4% (mean +3SD), respectively. When compared with a serum neutralizing antibody test (SNT) using chicken embryonated eggs, the rate of coincidence was 70.6% between the blocking ELISA and SNT, based on the titration of 20 duck DTMUV-positive serum samples.Conclusions/SignificanceThe blocking ELISA based on a neutralizing mAb allowed rapid, sensitive, and specific detection of neutralization-related antibodies against DTMUV.

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Development and validation of a monoclonal antibody blocking elisa for the detection of antibodies against both equine herpesvirus type 1 (EHV1) and equine herpesvirus type 4 (EHV4)

Cited by 16 publications

References 19 publications

Detection of Antibodies to Alphaviruses and Discrimination between Antibodies to Eastern and Western Equine Encephalitis Viruses in Rabbit Sera using a Recombinant Antigen and Virus‐specific Monoclonal Antibodies

Detection of Antibodies to Alphaviruses and Discrimination between Antibodies to Eastern and Western Equine Encephalitis Viruses in Rabbit Sera using a Recombinant Antigen and Virus‐specific Monoclonal Antibodies

Equine herpesvirus 1 and 4 infections: An update

Development of a Blocking ELISA for Detection of Serum Neutralizing Antibodies against Newly Emerged Duck Tembusu Virus

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