scholarly journals Elaboration of diagnostic means for detection antibodies to Newcastle disease virus

2017 ◽  
Vol 9 (1) ◽  
Author(s):  
Oksana Blotska
Keyword(s):  
Newcastle Disease Virus ◽  
Diagnostic Test ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Live Virus ◽  
Post Vaccination ◽  
Test Kit ◽  
Control Serum ◽  
Field Samples ◽  
The World

ObjectiveA test kit for the detection of antibodies to Newcastle disease virus(NDV) based on haemagglutination inhibition (HI) assay has beendeveloped and introduced into practice for the first time in Ukraine.IntroductionNewcastle disease (ND) is the most important infectious viraldisease of poultry. The world-wide economic loss from it is 2-3billion USD per year. ND is reportable to the World Organizationfor Animal Health (OIE). ND is caused by virulent strains of avianParamyxoviruses belonging to type 1. Industrial poultry farmingis rapidly developing in Ukraine. Ornithological fauna of Ukraineincludes about four hundred species of birds, 207 of which nestwithin its borders. The territory of Ukraine transits 3 out of 14transcontinental global migration flows. The wild birds are themain natural reservoir of ND agents. It is necessary to control theintensity of post-vaccination immunity in poultry and the timing ofrevaccinations. OIE recommends enzyme linked immunosorbentassays (ELISA) and HI test for these purposes [1]. However, it shouldbe noted that HI test, possessing high specificity and sensitivity, ismuch cheaper. Therefore, it is the excellent means for ND timelysurveillance.MethodsDuring the development of a new diagnostic kit, we used thereference strain “La-Sota”, which was obtained from the NationalCenter of Microorganism Strains of Ukraine. We have producedhaemagglutinating antigen using embryonated SPF fowl eggs and10-11 day incubation. A dilution of the virus was inoculated in0.1 ml volumes into the allantoic cavity and incubated at 35-37° Cfor 80-96 hours. For the purpose of NDV inactivation, we usedaminoethyleneimine at the final concentration of 0.1%. Positive serumwas prepared by immunizing 60-day-old chickens with live virus onceand by inactivated virus twice with an interval of 2 weeks. Negativeserum was obtained from healthy birds that did not contain antibodiesto NDV. The investigated blood sera were inactivated by heating(56 C/30 minutes). Samples of 1% suspension of chicken erythrocytesin phosphate buffered saline (pH 7.0-7.1) were used in HI tests.ResultsThe specific haemagglutination activity of the obtained antigenamounted to 10-11 log2.The test was performed using the 4HA unitsof the antigen. Positive control serum activity was in the range of 7-9log2. Negative control serum did not give results of more than 2 log2.The estimation of the quality indexes of the components of thediagnostic test-kit was performed using harmonized methods. In orderto examine sensitivity and specificity of HI test kits, antigens and serafrom commercial diagnostic kits were used. Also, certified negativecontrol serum and samples of International Standard sera were used,which were obtained from reference laboratories, namely againstthe following pathogens: Avian Influenza A (H5), Avian InfluenzaA (H7), Egg Drop Syndrome'76 Virus, Paramyxoviruses of 2 and3 serotypes, Reovirus, Avian Infectious Laryngotracheitis, AvianInfectious Bronchitis Virus, Mycoplasma gallisepticum, and NDV.In order to ensure a high degree of specificity for the antigen,special attention was given to the selection of a stabilizer for freeze-drying (the subject of a patent).Comparison between the national diagnostic test kit for HI andcommercial ELISA kit (IDEXX) in the evaluation of humoralimmune response to ND in vaccinated chickens was investigatedby examining of serum samples (n=152). Statistical analysis of datashowed that the correlation coefficient for the results of both tests was0.92. The relative sensitivity of HI test kit was 93.5% and the relativespecificity - 91.5%.The developed test kit was successfully used for the examinationof field samples. We developed regulatory documents, completed theprocedure of validation and registration in Ukraine of the commercialHI test kit for the detection of antibodies to NDV.ConclusionsThe use of the national standardized diagnostic test kit based onHI for detection of antibodies to NDV allows assessing the post-vaccination antibodies level that helps to maintain the disease-freestatus of the Ukrainian poultry industry with regard to ND.

Prevalence of Newcastle Disease Virus in Pakistan, its present status and future challenges

2021 ◽  
Vol 7 (2) ◽  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Current Status ◽  
Poultry Products ◽  
Cost Impact ◽  
The World ◽  
Future Challenges ◽  
The Mean ◽  
Major Industry

Newcastle disease is caused by Newcastle Disease Virus (NDV) leads to severe morbidity and mortality in poultry throughout the world and considered as lentogenic, mesogenic or velogenic based on the mean death of the chicken embryo. The NDV velogenic strain is deadly endemic in Pakistan. Poultry is considered as the second major industry in Pakistan having annual growth of 8-10%. Unfortunately, the increase of NDV cases leads to severe cost impact, loss of production and livelihood. This review highlights the current status and epidemiology of NDV in Pakistan. Various genotypes and sub-genotypes have been identified in Pakistan. Various ND cases have been reported in Pakistan which has very bad consequences on the economy and dealing of poultry products.

Ubiquitination on Lysine 247 of Newcastle Disease Virus Matrix Protein Enhances Viral Replication and Virulence by Driving Nuclear-Cytoplasmic Trafficking

2021 ◽  
Author(s):  
Tingyu Peng ◽  
Xusheng Qiu ◽  
Lei Tan ◽  
Shengqing Yu ◽  
Binghuan Yang ◽  
...  
Keyword(s):  
Viral Replication ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Nuclear Export ◽  
Viral Assembly ◽  
M Protein ◽  
Primary Function ◽  
The World ◽  
Export Signal

The Newcastle disease virus (NDV) matrix (M) protein is the pivotal element for viral assembly, budding and proliferation. It traffics through the cellular nucleus but performs its primary function in the cytoplasm. To investigate the biological importance of M’s nuclear–cytoplasmic trafficking and the mechanism involved, the regulatory motif nuclear export signal (NES) and nuclear localization signal (NLS) were deeply analyzed. Here, two types of combined NLS and NES signals were identified within NDV-M. The Herts/33-type M was found to mediate efficient nuclear export and stable virus-like particle (VLP) release, while the LaSota-type M was mostly retained in the nuclei and showed retarded VLP production. Two critical residues, 247 and 263, within the motif were identified and associated with nuclear export efficiency. We identified, for the first time, residue 247 as an important monoubiquitination site, the modification of which regulates the nuclear–cytoplasmic trafficking of NDV-M. Subsequently, mutant LaSota strains were rescued via reverse genetics, which contained either single or double amino acid substitutions that were similar to the M of Herts/33. The rescued rLaSota strains rLaSota-R247K, -S263R, and -DM (double mutation) showed about twofold higher HA titers and 10-fold higher EID 50 titers than wild-type (wt) rLaSota. Further, the MDT and ICPI values of those recombinant viruses were slightly higher than that of wt rLaSota probably due to their higher proliferation rates. Our findings contribute to a better understanding of the molecular mechanism of the replication and pathogenicity of NDV, and even those of all other paramyxoviruses. It is beneficial for the development of vaccines and therapies for paramyxoviruses. Importance Newcastle disease virus (NDV) is a pathogen that is lethal to birds and causes heavy losses in the poultry industry worldwide. The World Organization for Animal Health (OIE) ranked ND as the third most significant poultry disease and the eighth most important wildlife disease in the World Livestock Disease Atlas in 2011. The matrix (M) protein of NDV is very important for viral assembly and maturation. It is interesting that M proteins enter the cellular nucleus before performing their primary function in the cytoplasm. We found that NDV-M has a combined nuclear import and export signal. The ubiquitin modification of a lysine residue within this signal is critical for quick, efficient nuclear export and subsequent viral production. Our findings shed new light on viral replication and opens up new possibilities for therapeutics against NDV and other paramyxoviruses; furthermore, we demonstrate a novel approach to improving paramyxovirus vaccines.

scholarly journals Evaluation of Rapid Detection Kit against Avian Influenza A Virus and H5 Subtype for Field Sample

2017 ◽  
Vol 21 (1) ◽  
pp. 48 ◽  
Author(s):  
Michael Haryadi Wibowo ◽  
Tri Untari ◽  
Sidna Artanto ◽  
Krisdiana Putri ◽  
Surya Amanu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Avian Influenza ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Sensitivity And Specificity ◽  
Newcastle Disease ◽  
Rapid Test ◽  
Specificity Test ◽  
Test Kit ◽  
Reaction Test

Avian influenza virus is poultry viral disease, which causes high economic losses. Various efforts have been made to control the disease. One effort is required screening fast and precise diagnostic test. This study was aimed to determine the potential of rapid test kit of AIV/H5 Anigen Rapid Test for the detection of AI virus types A and subtype H5 in the field. Some tests were carried out, e.g.: the potential test, cross-reaction test, sensitivity and specificity test. Potency test was done to evaluate potential of detection limits of the kit, by having the test of serial dilution of AI virus positive control. Cross-reaction test was done to detect antigens other than AI virus H5N1, e.g.:  IB virus of Massachuset strain, IBV strain 4-91, Newcastle Disease virus, and Escherichia coli. Sensitivity and specificity test were applied to the filed samples which clinically and laboratory were confirmed as H5N1 positive. To confirm the result of rapid test was then being done by reverse transcriptase polymerase chain reaction. Based on these results it can be concluded that, Anigen Kit AIV/H5 Ag Rapid Test can detect antigen-containing samples having AI virus HA titer up to 26of type A virus, and up to 25 for subtype H5 virus. Anigen Kit AIV/H5 Ag Rapid Test showed no cross-reactions with Infectious Bronchitis virus, Newcastle Disease virus, and Escherichia coli. Anigen A Rapid Test Kit AIV Ag showed a sensitivity of 50% and specificity of 100%, while Anigen Ag Rapid Test Kit AIV/H5 showed a sensitivity of 25% and specificity is 100%.

scholarly journals Newcastle disease virus (NDV) expressing the spike protein of SARS-CoV-2 as a live virus vaccine candidate

EBioMedicine ◽  
2020 ◽  
Vol 62 ◽  
pp. 103132
Author(s):  
Weina Sun ◽  
Sarah R. Leist ◽  
Stephen McCroskery ◽  
Yonghong Liu ◽  
Stefan Slamanig ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Virus Vaccine ◽  
Newcastle Disease ◽  
Vaccine Candidate ◽  
Spike Protein ◽  
Live Virus ◽  
Live Virus Vaccine

Different Regimes for Challenge Instigation against Newcastle Disease Virus in Layer Birds

2018 ◽  
Vol 52 (1-4) ◽  
pp. 10-21
Author(s):  
MR Rahman ◽  
NA Rumi ◽  
MK Hossain ◽  
MS Rahman ◽  
MG Rahman ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Serological Test ◽  
Tap Water ◽  
Serum Antibody ◽  
Age Groups ◽  
Post Vaccination ◽  
Vaccination Programs ◽  
Antibody Titers

Newcastle disease is a highly contagious disease of poultry and causes huge economic loss in Bangladesh. In the study, the efficacy of different vaccination programs against Newcastle disease virus was performed to examine duration and level of antibody in two layer farms with different diluents. The experiment was carried out in Dinajpur district and serological test was performed in the Department of Microbiology, Hajee Mohammad Danesh Science and Technology University, Dinajpur during the period of July, 2017 to December, 2017. A total of 200 blood samples were randomly collected from different age groups of layer birds which were vaccinated with commercially available two live and one killed vaccine namely Avi ND LaSota, CEVAC NEW L and ITA-New (ND). Pre-vaccination serum antibody titers (Mean ± SD) of group A birds were 5.97±0.75. Firstly, birds were vaccinated with Avi ND LASOTA® vaccine, and CEVAC® NEW L vaccine. MASTERBLUE powder and ADVANCE NON-FAT were used as diluents in vaccination. After 40 days of post-vaccination, sera samples were procured and showed serum antibody titer (Mean ± SD) 7.0 ± 0.93. In case of farm-B, before vaccination antibody titers levels against NDV were Mean ± SD 5.85±0.75. Birds of farm-B were vaccinated with Avi ND LaSota® vaccine. After 40 days of post-vaccination, titer was Mean ± SD 6.7 ± 0.93. Then birds were vaccinated with ITA-New (ND) ® which were killed vaccine. After 40 days of post-vaccination, the titer was Mean ± SD 8.15± 0.93. MASTERBLUE powder and ADVANCE NON-FAT are suitable diluents to be used in vaccination against Newcastle disease because tap water may inactivate the vaccine due to its impurity in addition to containing higher level of chlorine and even the type of pipes or vessels used to distribute the drinking water. The results showed that the level of protection of vaccinated birds was satisfactory.

scholarly journals Sensitivity and Specificity of an In-house Sandwich ELISA Kit for Newcastle Disease Virus Antigen Detection

2013 ◽  
Vol 1 (2) ◽  
pp. 65-71
Author(s):  
Md Mostofa Kamal ◽  
Md Mortuza Ali ◽  
Md Ehsanul Haque ◽  
Mohammad Muradul Islam Chowdhury ◽  
Mohammad Aynul Haque ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Sensitivity And Specificity ◽  
Newcastle Disease ◽  
Sandwich Elisa ◽  
Antigen Detection ◽  
Test Method ◽  
Elisa Kit ◽  
Field Samples ◽  
Micro Organisms

Of all serological tests enzyme-linked immunosorbent assay (ELISA) is still considered the gold standard for the detection of antigens and antibodies of either macro or micro-organisms worldwide. The ELISA kits for serum antibody detection against many viruses and other micro-organisms of both man and animals are available in the market. Whereas, antigen detection ELISA kits for Newcastle disease virus (NDV) is not yet available in Bangladesh. The Present study was designed for the development of an economically feasible In-house Sandwich ELISA and to test its sensitivity and specificity for the detection of NDV antigens from clinically suspected field samples. 96- well flat bottom polystyrene plates coated with hyperimmune polyclonal serum against NDV raised in rabbits was used to capture NDV antigens. The anti-rabbit IgG and DAB with 30% H2O2 were used as conjugate and substrate respectively for standardization of the test method. The plate coated with serum diluted 10-3 was found suitable for capturing maximum antigen of NDV by the In-house Sandwich ELISA. The cut-off value of the present ELISA test was calculated as 0.855 and was able to capture the viral antigen present in the 10-4 fold dilution of allantoic fluid (AF) which is equivalent to 1HA unit, indicating the highest degree of sensitivity of the newly developed ELISA. In case of field samples, the newly developed ELISA kit was able to detect 100% viral antigens of NDV present in the feces, 95.50% of the brain tissue and oro-nasal swab and 94.12% of colon swab samples of either naturally and experimentally infected birds in this study. The ND virus specific polyclonal antibody used in the kit bind only with ND virus without any cross reactive antigens of other viruses of chicken like Avian influenza virus (AIV) and Infectious bursal disease viruses (IBDV). Therefore, findings of the present study clearly indicates that the newly developed In-house Sandwich ELISA kit can be used for rapid confirmatory diagnosis of Newcastle disease (ND) with minimum cost, using any kind of field samples from either sick or dead birds. DOI: http://dx.doi.org/10.3329/mh.v1i2.14093 Microbes and Health, 2012 1(2): 65-71

scholarly journals Detection of Newcastle Disease Virus (NDV) in Laughing Doves and the Risk of Spread to Backyard Poultry

2020 ◽  
Vol 64 (3) ◽  
pp. 1-12
Author(s):  
J. U. Okpanachi ◽  
J. U. Umoh ◽  
G. S. N. Kia ◽  
A. A. Dzikwi
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Fusion Gene ◽  
Haemagglutination Inhibition ◽  
Viral Disease ◽  
Poultry Production ◽  
Rt Pcr ◽  
Backyard Poultry ◽  
Field Samples

AbstractNewcastle disease (ND) is a highly infectious viral disease of birds caused by the Newcastle disease virus (NDV) and doves have been incriminated in previous outbreaks of the disease that have discouraged backyard poultry productions. This survey was done to detect and characterize the NDV from 184 swabs from the cloacae and pharynxes of 67 trapped laughing doves and 25 backyard poultry birds. The study utilized haemagglutination assay (HA) followed by haemagglutination inhibition (HI) tests on HA positive samples to screen field samples. Conventional reverse transcriptase polymerase chain reaction (RT-PCR) was conducted on the HI positives to characterize the NDV. This study revealed that of 134 dove samples screened, 88 (65.7 %) were HA positive. Of these HA positives subjected to HI testing, 37 (42.1 %) were HI positive. Interestingly, 21 (56.8 %) of the HI positives were also RT-PCR positive: 8 lentogenic, 12 velogenic, while one had both lentogenic and velogenic NDV. Comparatively, of the 50 chicken samples screened, 23 (46 %) were HA positive; and of these, HA positives subjected to HI testing, 16 (69.6 %) were HI positive. Only 4 (25 %) of the HI positives were RTPCR positive: 3 lentogenic and a velogenic NDV. From this study it was concluded that laughing doves were demonstrated to be infected with either lentogenic or velogenic NDV or both. The use of red blood adsorption-de-adsorption concentration of NDV enhanced the RT-PCR detection using the fusion gene primers NDV-F 4829 and NDV-R 5031. The detection of not only lentogenic but velogenic NDV in laughing doves poses a great risk to backyard poultry production.

scholarly journals The efficacy of binary ethylenimine-inactivated vaccines of Gianyar-1/AK/2014 virulent strain in protecting chickens against Tabanan-1/ARP/2017 virulent Newcastle disease virus isolates

2019 ◽  
Vol 12 (6) ◽  
pp. 758-764 ◽  
Author(s):  
Anak Agung Ayu Mirah Adi ◽  
I Nyoman Mantik Astawa ◽  
I Gusti Agung Arta Putra
Keyword(s):  
Antibody Response ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Post Vaccination ◽  
Group Iii ◽  
Group I ◽  
Virulent Newcastle Disease Virus ◽  
Inactivated Vaccines ◽  
The Mean

Aim: This study aimed to prepare binary ethylenimine (BEI)-inactivated virulent Newcastle disease virus (NDV) vaccine and to examine their ability to induce a protective antibody response in commercial chickens. Materials and Methods: A virulent NDV field isolate Gianyar-1/AK/2014 was propagated in chicken-embryonated eggs and was then inactivated with BEI at a concentration of 4 mM. Three groups of chickens with low-level (2 log2 hemagglutination inhibition [HI] units) maternally derived antibodies against NDV were then immunized with the BEI-inactivated vaccine. A commercial live vaccine (LaSota strain) was used as positive control, and phosphate-buffered saline (PBS) was used as negative control. A challenge experiment with a virulent NDV of Tabanan-1/ARP/2017 was performed at 3 weeks post-vaccination. Results: At 2 weeks post-immunization, the mean titers of antibodies against NDV in serum samples of chickens immunized with 0.2 mL of BEI-inactivated NDV (Group I), with live commercial NDV vaccine (Group II) and with PBS (Group III) were 3±0.94 log2 HI units, 4.9±0.99 log2 HI unit, and 0.0±0.0 HI units, respectively. At week 3 post-immunization, the mean titers of the antibodies for the three groups were 5±1.09 log2 HI units, 6.9±0.32 log2 HI units, and 0.00 HI units, respectively. The antibody titer induced by inactivated NDV Gianyar-1/AK/2014 isolates examined at 2 and 3 weeks post-vaccination was still at a significantly (p<0.01) lower level as compared to those induced by commercial life vaccine. However, the challenge test with virulent NDV of Tabanan 1/ARP/2017 isolates showed that all immunized chickens (Group I and II) survived without exhibiting any clinical sign post-challenge with the protection rates of 100%, whereas all chickens injected with PBS (Group III) died with clinical signs of ND. Conclusion: This finding shows that the BEI-inactivated vaccines prepared using virulent NDV of Gianyar-1/AK/2014 strain was able to induce protective antibody response in chickens but still at a lower level than those induce by commercial live NDV vaccine.

scholarly journals Humoral response of broilers to live Newcastle Disease virus vaccines manufactured by different companies

2020 ◽  
Vol 10 (3) ◽  
pp. 179-184
Author(s):  
Muhammad Danish Mehmood ◽  
Huma Anwar ◽  
Aamra Hashim ◽  
Muhammad Ismail ◽  
Sajjad Hussain
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Booster Dose ◽  
Haemagglutination Inhibition ◽  
Humoral Response ◽  
Post Vaccination ◽  
Vaccine Schedule ◽  
Vaccination Scheme ◽  
Time Periods

Increased incidence and severity of fatal Newcastle Disease Virus (NDV) in commercial and domestic poultry has been reported from across Pakistan. The present study was carried out to evaluate the efficacy of various live NDV vaccines (Gallivac, Intervet and Ceva) by adapting different vaccination schemes in broilers. Antigenic count of each vaccine and its generated antibody were determined by Haeamagglutination and Haemagglutination inhibition tests respectively. Two different NDV vaccination schemes were tested in such a way that one group had received three vaccines in different time periods whereas, the other after priming at 0 day, was followed by a single booster dose. For this purpose, healthy broilers were divided into four groups A, B, C, and D. On zero day of vaccination, there was no detectable anti NDV-HI titer for all vaccines (Gallivac=2, Intervet=2 & Ceva=2). In first vaccination scheme, detectable anti NDV-HI titer was observed on 16th day of vaccination (Gallivac=3.8, Intervet=4.2 & Ceva=3.6). All vaccines showed protective anti NDV-HI titer on 32nd day, post vaccination (Gallivac=5.8, Intervet=6.2, Ceva=5.8). In second vaccination scheme, detectable anti NDV-HI titer was achieved on 16th day of post vaccination (Gallivac=3.8, Intervet=4.2 & Ceva=3.6). While on 32nd day vaccination, all vaccines showed protective anti NDV-HI titer (Gallivac=5.4, Intervet=5.6 & Ceva=5.2). It is concluded that the two-dose vaccination program, with interval of 12 days, is much effective than the 3-dose vaccination scheme. However, in broilers, anti-NDV antibody titer was induced by all three types of vaccines manufactured by various companies. Keywords: Newcastle disease virus, Humoral Response, Haemagglutination inhibition test, Vaccine schedule

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