Standardization of an indirect enzyme linked immuno sorbent assay for measuring antibodies of infectious bursal disease virus (original) (raw)
Related papers
Journal of Virological Methods, 2010
The routine technique for detecting antibodies specific to infectious bursal disease virus is a serological evaluation by enzyme linked immunosorbent assay (ELISA) with preparations of whole virions as antigens. To avoid the use of complete virus in the standard technique, in-house VP2 and VP3 based ELISAs were developed. Accordingly, four types of indirect ELISAs viz., a commercial IDEXX-ELISA kit, VP2 and or VP3 antigen based ELISAs and a whole virus ELISA were compared with the virus neutralization test. It was concluded that the sensitivity and specificity at receiver-operating characteristics (ROC) optimized cut-off of four ELISAs viz., IDEXX-ELISA, VP2-ELISA and VP3-ELISA indicated similar performance whereas whole virus antigen based ELISA showed poor performance in comparison to other ELISAs. Similarly the positive and negative likelihood ratio of four ELISAs at an optimized cut-off indicated IDEXX-ELISA to be the best among all the four ELISAs while the performance of rVP3-ELISA and rVP2-ELISA is good as compared to the whole virus ELISA. Finally, the area under the ROC curve (AUC) of four ELISAs which represented a summary statistics of the overall diagnostic performance of the test also indicated that the IDEXX-ELISA, VP3-ELISA and VP2-ELISA had similar and relatively better performance when compared to whole virus antigen-ELISA.
Journal of Agricultural Science and Technology A, 2019
A study on infectious bursal disease virus (IBDV) on chickens of Cobb-500 strain broiler breed at Thakurgaon district of Bangladesh was performed. The protective antibody was measured on one day old chicks (DOC) and post-vaccinated (PV) flocks up to 75 weeks by indirect enzyme linked-immunosorbent assay (I-ELISA). The assays have included five flocks with vaccination historic against IBDV contained 45,000 birds with age ranging from DOC to 75 weeks, just before culling. Maternally derived antibody (MDA) mean titer (MT) ranged from 3,395 to 5,184. The antibody from serum samples (N = 92 per flock) were titer tested by I-ELISA at age 14 d, 5, 8, 23, 50 and 75 weeks of each vaccinated flock. Antibody titer level gradually decreased before vaccination. Vaccination done by intermediate plus vaccine resulting titers level was increased and stayed at the same level. The antibody MT at the 14th day was 500, which supported Deventer method. The protective antibody MT was declined at growing, laying, mid laying and last stage of laying groups. So MDA titer was enough in offspring that could protect birds easily.
This study was designated to investigate the antibody response of broiler chickens against eight commercial IBD live vaccines. A total of 460 one-day Ross broiler chicks were divided to 9 groups, eight groups were vaccinated with IBD live vaccines and the last one was served as control. Four groups were vaccinated with intermediated vaccines, whereas the other fourth were given intermediated plus vaccines. All vaccinated groups were administrated at 14 th day of age via drinking water route. Maternal derived antibody (MDA) and post-vaccination antibody response were tested by ELISA. Blood samples were collected at one day old and at 21 st , 28 th and 35 th day of age post-vaccination. Indirect ELISA test revealed that the mean of maternal derived antibody was 4852±745. Significant differences (P<0.05) among means of antibody titers of all vaccinated groups were found at 21 st ,28 th and 35 th day of age compared with that of control group. The results also showed that groups which were vaccinated with intermediated plus vaccines(E and H vaccines) exhibited high level of antibody especially groups 5 and 8 than those which vaccinated with intermediate vaccines. In conclusion, Intermediate plus vaccines induced higher antibody titers than other vaccines, although some intermediate vaccines induced similar titers of antibody .E and H vaccines which were administered to groups 5 and 8 respectively induced better antibody titers.
2018
Infectious Bursal Disease (IBD) is one of the oldest and widely known poultry diseases all over the world. It is caused by IBD virus of Avibirnavirus genus of family Birnaviridae family. IBD has great economical impact on backyard poultry world as it causes high weakness and mortality. The objective of present study was to evaluate the antibody response in village chickens in India after vaccinating them with IBD live vaccine. Serum was collected at regular intervals from chickens up to 112 days after vaccination. Antibodies against IBD virus were measured using ELISA method. It was observed that the vaccines with both intermediate and intermediate invasive strain caused good immune response in the birds. Serum antibody level was found significantly high in 28 days blood collection, which decreased gradually up to 112 days. It was also observed that Intermediate invasive strain produced higher amount of antibodies than intermediate strain of the vaccine. Furthermore, it is also sugg...
Indian Journal of Animal Research, 2021
Background: In recent years, Infectious bursal disease is continuously occurring even after vaccination in India and requires an inclusive diagnosis. Therefore, the present study was undertaken to diagnose IBD through molecular and culture methods. Methods: One pooled sample, from each of 54 flocks having birds with IBD like symptoms, was collected. History of bird type, age and vaccination was recorded. Samples were subjected to RT-PCR, egg embryo culture and chicken fibroblast cells culture. Result: A total of 49669 out of 517900 (9.59 %) of birds, aging 3-6 weeks, were displaying the signs similar to IBD.In RT-PCR, 21 (38.88%) samples were found positive which belonged to11 (52.38%) vaccinated and 10 (47.62%) unvaccinated flocks.The RT-PCR positive samples were successfully cultivated for the virus through egg embryo and cell culture. The CEF culture was found least sensitive compared to egg embryo culture and RT-PCR.
Belonging to genus Avibirnavirus and family Birnaviridae infectious bursal disease virus (IBDV) is a double stranded RNA virus and it causes an acute highly infectious disease in poultry resulting in watery diarrhoea, anorexia, high morbidity and mortality and hemorrhagic lesions on breast and leg muscles leading to down grading of poultry meat. The present study was designed to isolate and molecularly identify the causative agent (IBDV) from a clinically suspected flock of infectious bursal disease and to check the effects of isolated virus on cell mediated immune response and serum biochemical parameters in broilers along with reference strain (IBDV-2512). Bursae were collected and subjected to trituration and supernatent when inoculated in 9-day old embryonated chicken eggs resulted in the death of all the embryos during first three blind passages. Every triturate produced a clear and distinct line of precipitation with IBDV-known antisera in agar gel precipitation test (AGPT). Serum samples collected at the time of occurrence of disease presented a low anti-IBDV titer which was between 1:2 and 1:8 as elucidated by indirect haemagglutination inhibition (IHA) test while serum samples of same flock collected 14 days after first sampling presented a drastic increase in anti-IHA-IBDV antibodies that was between 1:64 and 1:512. Reverse transcriptase polymerase chain reaction (RT-PCR) revealed a product of approximately 743bp of VP2 gene of IBDV for all three suspected samples along with the reference strain. Broiler birds of 3 weeks of age when injected with field isolated resulted in decreased lymphoproliferative response as elucidated by tuberculin test and serum biochemical parameters were also altered in field isolated injected birds and these alterations were more or less similar to that of birds injected with reference strain (IBDV-2512) suggesting high pathogenicity of isolated virus.
The current study was conducted to evaluate the pathogenicity and immunosuppressive effects of GM-97 strain of infectious bursal disease virus in commercial broiler chickens. A total of 500 broiler chickens were vaccinated with the virus through oral route at 10 and 17 days of age (10 2-10 3 EID50/dose). Chickens were also vaccinated with Newcastle disease virus (Hitchner B1) orally at 14 and 21 days old. Chickens were euthanized (at 12, 14, 16, 20, 23, 26 days of age) after measuring body weight. Bursa of Fabricius was examined for any gross lesion, weighed and processed for histological investigations. Bursa to body weight ratio and bursal lesion scoring were made to evaluate pathogenicity of the virus. Blood samples were analyzed for antibody response to ND vaccine virus using HI test. Results showed that the GM-97 strain of IBDV induced mild to moderate depletion of lymphoid cells in the center of bursal follicles and non-significant difference in bursa to body weight ratio amongst vaccinated and unvaccinated chickens. Chickens responded well to ND vaccine by mounting high level of serum NDV specific HI antibody titers. It can be concluded from the present study that GM-97 strain of IBDV has mild pathogenicity but is not immunosuppressive.
IBDV Gm 11 (Simbios eleven-molecular group) has been detected since 1997 in many farms of commercial broilers and layers causing high mortality (2 to 15%) and severe macro and microscopic damage in cloacal bursae, spleen, thymus, kidney and liver. Five serial passages of 2050/ 97-Gm 11 IBDV sample by CAM route in SPF chickens embryonated eggs did not elicit increased embryo mortality. High mortality (100%) of 21 day-old SPF leghorn chickens and severe bursal and splenic lesions were seen from 24 up to 48 hours after eye-drop inoculation of 2050/ 97 strain (50 mL of 10-2 dilution of 10% bursae homogenate). Mortality was not detected when vaccinated SPF and broiler chickens were inoculated. One dead bird was found among ten challenged unvaccinated broilers. Variations in the intensity of cloacal bursae injury and spleen response were found between unvaccinated and vaccinated broiler chickens. IBDV antibodies were detected by ELISA test in almost all vaccinated SPF chickens before challenge while low number of commercial vaccinated and unvaccinated broilers were serologically positive (0 to 3 birds in 18). Increasing IBDV antibody titers were detected after challenge with 2050/97 strain and highest GMTs were found in broilers. It was concluded that 2050/97 strain is a highly virulent IBDV and SPF leghorn chickens immunized with BV8 intermediate vaccine strain were resistant to the challenge. Increasing susceptibility was found from experimental groups of unvaccinated broilers to vaccinated broilers and to unvaccinated SPF birds. It is discussed that passive immunity was involved in the rate of protection of challenged unvaccinated broiler and in the immune response impairment after vaccination of broilers chicks. The use of a constant virus suspension with known potency to challenge the experimental birds was suitable to evaluate vaccination efficacy. Evaluation of bursal and splenic responses at early and delayed time after challenge were useful to estimate vaccination efficacy and field interactions.
African Journal of Microbiology Research, 2012
Infectious bursal disease is one of the most important killer viral disease of poultry. This study was designed for potency testing of immunized yolk in infectious bursal disease (IBD) virus infected birds and to purify yolk immunoglobulin from egg yolk of immunized hens. Twenty commercial layers were raised in poultry shed of the University and were vaccinated with oil based killed Gumboro vaccine twice at an interval of 15 days at the age of 26 weeks interval to get immuned yolk. Eggs were collected at two weeks interval till two and half months after boosting. Immune yolks were purified by chemical means. Antibody titer against IBD in egg yolk and semi purified egg yolk IgY was measured by indirect ELISA kit method. Fifty commercial chicks of 15 days old were divided into eight groups and were reared in poultry shed in the University up to 36 days. The birds were challenged at 30th day and they were provided with passive therapy of immune yolk and semi purified IgY after 3 days of challenge. The birds which received semi purified immune yolk and antibody titre having more or less 3000 showed 20% mortality in each group. In conclusion semi purified IgY and immune yolk having titer more than 4000 were relatively effective in prevention of infectious bursal disease virus infected birds and protective antibody titer remain in yolk for two and half months after killed vaccination.