Concanavalin A- Sandwich-ELISA for the detection of antibodies against Newcastle disease virus (NDV) in chicken

2019 ◽  
Author(s):  
T.R. Kannaki ◽  
E. Priyanka ◽  
Santosh Haunshi
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Concanavalin A ◽  
Newcastle Disease ◽  
Indirect Elisa ◽  
Sandwich Elisa ◽  
Haemagglutination Inhibition ◽  
Con A ◽  
Serum Samples ◽  
Serum Dilution

Concanavalin A (Con A), a lectin interacts with carbohydrate moieties of viruses and provide stable and sensitive detection when used as a capture agent. Indirect ELISA methods need purified Newcastle disease virus (NDV) or recombinant antigens for adsorption, whereas use of Con A as capture agent will enable the use of non-purified and non-concentrated virus as antigen replacing costly and time-consuming virus purification step. Con A based sandwich ELISA with non-purified NDV whole virus antigen with single serum dilution format has been developed in this study. The optimum concentrations of the capture agent, Con A and non-purified antigen preparations were determined by checker-board titration. Briefly, microplates were coated with predetermined optimum concentration of ConA (0.5 mg/ml; 50µg per well) and incubated for 18h at 4°C. After washing, allantoic fluid with Newcastle disease virus (NDV) LaSota (HA titre, 210) at a constant predetermined dilution (1:1; 50µl) was coated and incubated for 45 min at 37°C, followed by blocking with 2 % bovine serum albumin for 45 min at 37°C. The antigen coated plates were used in the detection of antibody titre against NDV in serum samples at single serum dilution of 1: 500. Then, wells were added with goat anti-chicken IgG horseradish peroxidase conjugate and incubated for 1h at 37°C, followed by addition of TMB substrate and the plates were read spectrophotometrically at 450 nm. ELISA antibody titres were determined by standard serial dilution of positive sera and endpoints were calculated by a subtraction method. By using positive negative threshold curve (PNT), intercept and slope of the standard curve were calculated. Total of 271 random chicken serum samples were analyzed for antibodies against NDV by Haemagglutination inhibition assay (HI), indirect ELISA and compared with the Con A- S- ELISA developed in this study. The Con A-S-ELISA showed a high coefficient of correlation (r=0.85, n=271, P less than 0.01) and an agreement of ê=0.99 with the commercially available Indirect-ELISA. The relative sensitivity and specificity were 98% and 85% respectively in comparison to HI test. Hence, the Con A-S-ELISA is a simple, easy and effective for monitoring serum antibody levels against NDV.

scholarly journals Determination of serum antibody titres and immune status of layer flocks against Newcastle Disease virus at Chittagong district of Bangladesh

1970 ◽  
Vol 1 (2) ◽  
pp. 35-38 ◽  
Author(s):  
MA Kashem ◽  
M Parvej ◽  
MA Hashem ◽  
MM Moula ◽  
ASMG Kibria
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Immune Status ◽  
Haemagglutination Inhibition ◽  
Serum Antibody ◽  
Age Groups ◽  
Serum Samples ◽  
Specific Immunity ◽  
Antibody Titres

A study was conducted to assess the level of serum antibody titres and immune status of layer birds against Newcastle Disease virus by Haemagglutination Inhibition (HI) test in different areas of Chittagong district during November to December, 2010. Sixteen layer flocks were selected based on different ages of birds. A total of 235 serum samples were collected and tested at Microbiology laboratory of CVASU. HI test was performed using commercial Newcastle Disease vaccine (Avinew®) as a source of 4HAU virus antigen. The antibody titre (GMT) levels in 18-26 weeks age group were found to be 70.198, followed by 47.551, 34.776, 17.281 and 18.855 in 27-40, 41-57, 58-73 and >73 weeks age groups, respectively. Moreover, 100% specific immunity against ND was found in 18-26, 27-40 and 41-57 weeks age groups of birds, whereas 93.33 and 94.73% specific immunity was found in 58-73 and >73 weeks age groups, respectively. On an average, 97.87% layer birds showed specific immunity and 2.13% showed nonspecific immunity against NDV. We considered HI titre of 1:8 or above as specific immunity and less than 1:8 as non specific immunity. Highest HI titre was found at the age of 18-26 weeks and lowest titre was at 58-73 weeks of age. The lower level of HI titre seemed to be directly related to some important factors relating to vaccination which have been highlighted in this paper. Key words: Antibody titers; Immune status; HI test; Newcastle disease virus; Layer birds. DOI: http://dx.doi.org/10.3329/ijns.v1i2.8818 International Journal of Natural Sciences (2011), 1(2):35-38

scholarly journals Comparative Performance of In-House and Commercially Available ELISA Kits for the Detection of Antibody Against Newcastle Disease Virus Vaccine

2013 ◽  
Vol 22 (1-2) ◽  
pp. 55-64
Author(s):  
MMI Chowdhury ◽  
MT Islam ◽  
A Aktar ◽  
MKJ Bhuiyan ◽  
MM Kamal ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Sensitivity And Specificity ◽  
Newcastle Disease ◽  
Indirect Elisa ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Dilution ◽  
Positive Correlation ◽  
Elisa Kit

An In-house Indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of serum antibody titre against Newcastle disease virus (NDV) and was compared its sensitivity and specificity with the commercially available NDV antibody detection ELISA kit (Biocheck®, USA). The reference NDV was purified by centrifugation, ultracentrifugation and by sucrose density gradient ultracentrifugation. This purified NDV was used for coating of 96-well flat bottomed microtitre plate and to raise hyperimmune sera (known) in Fayoumi chickens. In the standardization test, the antigen dilution of 10-6 and the serum dilution of 10-3 were considered to be optimum for the present ELISA system. The correlation regression analysis was performed to construct a standard curve equation where a good positive correlation was observed (r = 0.912, n = 8, P<0.01). The equation was used to convert corrected absorbance readings of the single working dilution (1 : 1000) directly into predicted ELISA antibody activity titres. In the sensitivity and specificity test, the serum dilution of 10-5 appeared to be the highest dilution which had the maximum lowest capacity to bind with the coated antigen of the present ELISA kit and only anti-NDV serum was found to bind with the coated antigen instead of serum of IBDV in the plate which revealed the high specificity of the developed In-house Indirect ELISA kit. In a comparative study with the 80 chicken sera samples, a significant positive correlation (r = 0.901, n = 80, P<0.01) was found between In-house Indirect ELISA and commercial ELISA kit (Biocheck®, USA).DOI: http://dx.doi.org/10.3329/pa.v22i1-2.16467 Progress. Agric. 22(1 & 2): 55 - 64, 2011

scholarly journals The Pathology of Vaccination of Chickens with Varying Doses of Lentogenic LaSota Strain of Newcastle Disease Virus

2021 ◽  
Vol 41 (1) ◽  
pp. 62-72
Author(s):  
A.O. Igwe ◽  
M.E. Sanda ◽  
U.E.I. Nnsewo ◽  
C.J. Okonkwo ◽  
O. Onyebgula
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Broiler Chickens ◽  
Haemagglutination Inhibition ◽  
Clinical Signs ◽  
Lymphoid Hyperplasia ◽  
Post Inoculation ◽  
Serum Samples ◽  
Antibody Titres

Recently, it was demonstrated under laboratory conditions that increased doses of LaSota vaccine increased ND antibody response significantly in chickens. In this study, we have used the same model to investigate whether vaccination with increased doses of  lentogenic LaSota strain of Newcastle disease virus are associated with pathological changes in chickens. Four-week-old broiler chickens (n=100) were randomly assigned into four groups of 25 each: ZD, each drenched with phosphate-buffered saline, SD, DD and TD broilers were each drenched with single, double and triple dose of LaSota vaccine, respectively. The chickens were observed for clinical signs and lesions. Serum samples were collected from the chickens in all the groups at weekly intervals post inoculation (PV) and assayed for haemagglutination inhibition (HI) antibodies. The vaccinated broilers showed no morbidity and mortality. Only the bursa of all the vaccinated groups appeared slightly reduced in size on day 10 PV. The histopathological changes were lymphoid  hyperplasia and formation of germinal centres in the spleen and caecal tonsils from days 3 to 6 PV and mild depletion of bursal lymphocytes on day 10 PV. Generally, the integrity of the lymphoid organs was intact. Groups DD and TD antibody titres were  significantly (P < 0.05) higher than that of the SD on day 21 PV. This suggests that increased doses of LaSota vaccine does not cause  pathologic impairment and may be considered in improving the performance of the vaccine in the control of velogenic ND. Key words: Newcastle disease, LaSota vaccine, pathology, broiler chickens

scholarly journals Spatial Distribution and Seroprevalence of Newcastle Disease in Kaduna State, Nigeria

2021 ◽  
Vol 65 (1) ◽  
pp. 37-44
Author(s):  
W. A. Adanu ◽  
J. U. Umoh ◽  
J. Kabir ◽  
J. K. P. Kwaga ◽  
G. R. Otolorin ◽  
...  
Keyword(s):  
Spatial Distribution ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Haemagglutination Inhibition ◽  
Disease Prevalence ◽  
Poultry Production ◽  
Serum Samples ◽  
System A ◽  
Entry Points

Abstract Newcastle disease is one of the greatest constraints to the development of poultry production in Nigeria. In this study, the spatial distribution of Newcastle disease antibodies was determined using the Geographic Information System. A total of 400 serum samples were collected from chickens in districts around Kaduna Metropolis and screened for Newcastle disease virus antibodies using the haemagglutination inhibition test done according to the procedure of OIE (2002). The spatial distribution demonstrated that the highest antibody titre level for Newcastle disease was closely associated with communities that were at entry points (Zaria— Kaduna road; Nnamdi Azikiwe bypass road; Television garage; Abuja—Kaduna expressway and Kachia road) to the metropolis and houses that are closely situated near live bird markets signifying the importance of bird movements in the spread of the disease. About 31 % (124 of 400) of chickens had antibodies to Newcastle disease virus, with exotics breeds (32.0 %) with more Newcastle disease virus antibodies than local breeds (29.8 %). The Newcastle disease prevalence was 33.9 % (39 out of 115), 31.8 % (41 out of 129) and 28.2 % (44 out of 156) for Kaduna North, Kaduna South and Chikun Local Government Areas (LGA), respectively. A Newcastle disease prevalence of 29.8 % (54 out of 181) was recorded in the local chickens, while 32.0 % (70 out of 219) was obtained from the exotic chickens.

scholarly journals Evaluation of Antibody Production against Newcastle Disease Virus after Immunization with Different Vaccines in Fayoumi Chicks

1970 ◽  
Vol 25 (1) ◽  
pp. 31-35
Author(s):  
Ahama Ranjan Barua ◽  
M Mansurul Amin ◽  
Shaiful Islam ◽  
Sukanta Chowdhury ◽  
M Shafiqul Islam Khan ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Antibody Production ◽  
Newcastle Disease ◽  
Research Work ◽  
Haemagglutination Inhibition ◽  
Serum Samples ◽  
Strain Group ◽  
Control Serum ◽  
Group B

The research work was performed in the experimental sheds and virology laboratory of the Department of Microbiology and Hygiene, Faculty of Veterinary Science, Bangladesh Agricultural University (BAU), Mymensingh during the period from January to November 2006 for the detection of persistence of maternallyderived antibody (MDA) against Newcastle disease virus (NDV) in Fayoumi chicks as well as evaluation of antibody production in chicks after vaccination. Five experimental groups of 110-day-old Fayoumi chicks were vaccinated for comparative study. Group A was vaccinated with baby chick ranikhet disease vaccine (BCRDV, F-strain), group B with Izovac B1 Hitchner® (B1 strain), group C with Cevac New L® (LaSota strain) and group D with Avinew® (VG/GA strain) through intraocular (i/o) route while group E was kept as unvaccinated control. Serum samples obtained from 10 randomly selected birds from each group on the occasion of 7, 13, 15, 17, 19 and 21 days post-vaccination (DPV) were subjected to haemagglutination inhibition (HI) test. It was found that persistence of MDA against Newcastle disease virus (NDV) retained until the age of day 20 in Fayoumi chicks and among the four commercial vaccines, Avinew® (VG/GA strain) appeared to be slightly superior compared to those of BCRDV (F-strain), Izovac B1 Hitchner® (B1 strain) and Cevac New L® (LaSota strain) in respect of HI antibody response. The F-values of HI titre among four vaccines were significantly different (p <0.01). Keywords: Newcastle disease virus (NDV); Fayoumi chicks; Maternally-derived antibody (MDA); Haemagglutination inhibition (HI); Newcastle disease (ND) vaccinesDOI: http://dx.doi.org/10.3329/bjm.v25i1.4852 Bangladesh J Microbiol, Volume 25, Number 1, June 2008, pp 31-35

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