An Enzyme Linked Immunosorbent Assay (ELISA) for Detection of Antibodies to the Koi Herpesvirus (KHV) in the Serum of Koi Cyprinus carpio

Adkison, Mark A.; Gilad, Oren; Hedrick, Ronald P.

doi:10.3147/jsfp.40.53

Cited by 79 publications

(94 citation statements)

References 28 publications

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“…Pooled sera of 10 wild common carp diagnosed CyHV-3 positive by PCR were used as a positive control and pooled sera of 10 cultured common carp never exposed to CyHV-3 were used as a negative control. Before the analysis, ELISA was optimized using the control sera diluted to 1:2500 or higher, because the extent of cross-reaction with anti-cyprinid herpesvirus 1 antibodies is reduced at those dilutions (Adkison et al, 2005). The optimal ELISA reagent concentrations and serum dilution producing the highest positive-to-negative (P/N) ratio were determined using checkerboard titrations and were subsequently used for further experiments.…”

Section: Measurement Of Serum Anti-cyhv-3 Antibodiesmentioning

confidence: 99%

“…One-half each of a 96-well microplate (Costar 3596; Corning, NY, USA) was coated with 50 ml per well of 2 mg ml À1 purified CyHV-3 (Adkison et al, 2005) in bicarbonate buffer (pH 9.6) or 50 ml per well of bicarbonate buffer for 1.5 h at 37 1C. The plates were blocked with 300 ml per well of blocking buffer for 1 h. After three washes with Tris-buffered saline containing Tween-20, 50 ml per well of the serum samples diluted to 1:2500 was added in duplicate to the antigen-coated and -uncoated wells to correct nonspecific binding for each serum sample.…”

Section: Measurement Of Serum Anti-cyhv-3 Antibodiesmentioning

confidence: 99%

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Transmission dynamics of an emerging infectious disease in wildlife through host reproductive cycles

et al. 2010

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Emerging infectious diseases are major threats to wildlife populations. To enhance our understanding of the dynamics of these diseases, we investigated how host reproductive behavior and seasonal temperature variation drive transmission of infections among wild hosts, using the model system of cyprinid herpesvirus 3 (CyHV-3) disease in common carp. Our main findings were as follows: (1) a seroprevalence survey showed that CyHV-3 infection occurred mostly in adult hosts, (2) a quantitative assay for CyHV-3 in a host population demonstrated that CyHV-3 was most abundant in the spring when host reproduction occurred and water temperature increased simultaneously and (3) an analysis of the dynamics of CyHV-3 in water revealed that CyHV-3 concentration increased markedly in breeding habitats during host group mating. These results indicate that breeding habitats can become hot spots for transmission of infectious diseases if hosts aggregate for mating and the activation of pathogens occurs during the host breeding season.

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Section: Measurement Of Serum Anti-cyhv-3 Antibodiesmentioning

confidence: 99%

Section: Measurement Of Serum Anti-cyhv-3 Antibodiesmentioning

confidence: 99%

Transmission dynamics of an emerging infectious disease in wildlife through host reproductive cycles

et al. 2010

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“…A progressive increase in anti-KHV antibodies in naturally exposed koi is seen by ELISA [69]. These antibodies are detected in the serum at 3 weeks after experimental infection and in survivors after 1 year following a natural infection [1,39,78,81]. KHV infection induces a high prevalence (54 %) of KHV specific antibodies in surviving wild fish population [83]; while studies in farmed fish of England show almost 85-93 % of fish are seropositive by ELISA, even 1 year after disease outbreak [81].…”

Section: Detection Of Khv Antigen/antibodiesmentioning

confidence: 95%

“…However, cross-reactivity with related cyprinid herpes virus or non-viral protein is a potential problem with immunostaining. ELISA is also useful in detection of KHV antigen from infected tissues or fish droppings [1,16]. Detection of specific antibodies in serum by ELISA is an indirect method for diagnosis of KHV.…”

Section: Detection Of Khv Antigen/antibodiesmentioning

confidence: 99%

Koi Herpes Virus: A Review and Risk Assessment of Indian Aquaculture

et al. 2012

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Common carp (Cyprinus carpio) is a widely cultivated freshwater fish for human consumption, while koi carp, is a farmed colored sub species of common carp used for ornamental purposes. Since 1998, both common carp and koi carp are severely affected by a viral disease called as Koi herpes virus disease (KHVD). This disease is caused by Koi herpes virus (KHV), also known as cyprinid herpes virus-3. The virus causes interstitial nephritis and gill necrosis in carps, so it is also termed as carp interstitial nephritis and gill necrosis virus. KHV is a double stranded icosahedral DNA virus belonging to family Alloherpesviridae, with a genome size of 295 kbp, larger than any member of Herpesviridae. The viral genome encodes 156 potential protein coding open reading frames. Each virion consists of forty structural proteins, which are classified as capsid (3), envelope (13), tegument (2) and unclassified (22) structural proteins. Diagnosis of KHVD is mainly based on detection of viral DNA by polymerase chain reaction amplification using specific primers or loop mediated isothermal amplification. Temperature dependent latent infection is unique to KHV; and carrier fish are often not detected, thereby possibly resulting in spread of this pathogen to newer areas. The disease is now known to occur in, or has been recorded from at least 26 different countries of the world. Fortunately, KHVD has not been reported from India or from Indian major carps. To monitor the disease status of the country, a total of 254 fish samples collected from different parts of India were screened by PCR for the presence of KHV. None of the tested samples were found to be positive for KHV. These results demonstrate that tested samples from different parts of India were apparently free from KHV. Preliminary risk assessment of KHV suggest that in the event of unrestricted importation of koi carps into our country, there is a higher probability of risk to aquaculture as compared to natural waters. So there is strong need to develop diagnostic capabilities and launch surveillance programmes for KHV in India.

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“…The KHV antibody enzyme-linked immunosorbant assay (ELISA) was prepared in combination of the assays according to [34,35] with some modifications. Briefly, the Medisorp ELISA plate (Nunc) was used for coating the plate with 3 µg purified KHV/ml [36], blocked with Roti Block ® (Roth), and as secondary antibody a monoclonal anti-carp IgM (F 16, Aquatic Diagnostics) at a dilution 1:64 and the anti-trout IgM monoclonal antibody 4C10 at a dilution of 1:1000 were used.…”

Section: Indirect Methods Of Khv Detection In Serum (Plasma) Samplesmentioning

confidence: 99%

Is There Any Species Specificity in Infections with Aquatic Animal Herpesviruses?–The Koi Herpesvirus (KHV): An Alloherpesvirus Model

Bergmann¹,

Cieslak²

2016

Fish Aquac J

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An Enzyme Linked Immunosorbent Assay (ELISA) for Detection of Antibodies to the Koi Herpesvirus (KHV) in the Serum of Koi Cyprinus carpio

Cited by 79 publications

References 28 publications

Transmission dynamics of an emerging infectious disease in wildlife through host reproductive cycles

Transmission dynamics of an emerging infectious disease in wildlife through host reproductive cycles

Koi Herpes Virus: A Review and Risk Assessment of Indian Aquaculture

Is There Any Species Specificity in Infections with Aquatic Animal Herpesviruses?–The Koi Herpesvirus (KHV): An Alloherpesvirus Model

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