@article{Blotska_2017, title={Elaboration of diagnostic means for detection antibodies to Newcastle disease virus}, volume={9}, url={https://ojphi.org/ojs/index.php/ojphi/article/view/7681}, DOI={10.5210/ojphi.v9i1.7681}, abstractNote={<div style="left: 78.2727px; top: 292.008px; font-size: 12.3207px; font-family: sans-serif; transform: scaleX(1.0823);" data-canvas-width="55.467823232323234">Objective</div><div style="left: 91.3182px; top: 305.011px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.990052);" data-canvas-width="328.8951148989899">A test kit for the detection of antibodies to Newcastle disease virus</div><div style="left: 78.2727px; top: 319.506px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.960167);" data-canvas-width="343.2918611111111">(NDV) based on haemagglutination inhibition (HI) assay has been</div><div style="left: 78.2727px; top: 334.001px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00332);" data-canvas-width="333.62010606060585">developed and introduced into practice for the first time in Ukraine.</div><div style="left: 78.2727px; top: 361.584px; font-size: 12.3207px; font-family: sans-serif; transform: scaleX(1.11884);" data-canvas-width="71.86668434343434">Introduction</div><div style="left: 91.3182px; top: 374.587px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.989998);" data-canvas-width="330.9563691919192">Newcastle disease (ND) is the most important infectious viral</div><div style="left: 78.2727px; top: 389.081px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.984364);" data-canvas-width="344.1173484848485">disease of poultry. The world-wide economic loss from it is 2-3</div><div style="left: 78.2727px; top: 403.576px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.957138);" data-canvas-width="343.3731777777776">billion USD per year. ND is reportable to the World Organization</div><div style="left: 78.2727px; top: 418.071px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.03374);" data-canvas-width="343.0454469696971">for Animal Health (OIE). ND is caused by virulent strains of avian</div><div style="left: 78.2727px; top: 432.566px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.998756);" data-canvas-width="344.4376868686868">Paramyxoviruses belonging to type 1. Industrial poultry farming</div><div style="left: 78.2727px; top: 447.061px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.969093);" data-canvas-width="343.4951527777778">is rapidly developing in Ukraine. Ornithological fauna of Ukraine</div><div style="left: 78.2727px; top: 461.556px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.986665);" data-canvas-width="344.3625305555555">includes about four hundred species of birds, 207 of which nest</div><div style="left: 78.2727px; top: 476.051px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.994373);" data-canvas-width="344.35144191919215">within its borders. The territory of Ukraine transits 3 out of 14</div><div style="left: 78.2727px; top: 490.546px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.02013);" data-canvas-width="344.8565909090904">transcontinental global migration flows. The wild birds are the</div><div style="left: 78.2727px; top: 505.041px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.949073);" data-canvas-width="343.5801656565656">main natural reservoir of ND agents. It is necessary to control the</div><div style="left: 78.2727px; top: 519.536px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.03066);" data-canvas-width="343.072552525252">intensity of post-vaccination immunity in poultry and the timing of</div><div style="left: 78.2727px; top: 534.031px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.0105);" data-canvas-width="344.3120156565659">revaccinations. OIE recommends enzyme linked immunosorbent</div><div style="left: 78.2727px; top: 548.526px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.984061);" data-canvas-width="341.862659090909">assays (ELISA) and HI test for these purposes [1]. However, it should</div><div style="left: 78.2727px; top: 563.021px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.04511);" data-canvas-width="343.10828257575776">be noted that HI test, possessing high specificity and sensitivity, is</div><div style="left: 78.2727px; top: 577.516px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.963239);" data-canvas-width="343.74772727272733">much cheaper. Therefore, it is the excellent means for ND timely</div><div style="left: 78.2727px; top: 592.011px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00155);" data-canvas-width="62.613833333333325">surveillance.</div><div style="left: 78.2727px; top: 619.594px; font-size: 12.3207px; font-family: sans-serif; transform: scaleX(1.07424);" data-canvas-width="50.65042676767676">Methods</div><div style="left: 91.3182px; top: 632.597px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.985845);" data-canvas-width="331.2767075757576">During the development of a new diagnostic kit, we used the</div><div style="left: 78.2727px; top: 647.092px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.969406);" data-canvas-width="343.60603914141416">reference strain “La-Sota”, which was obtained from the National</div><div style="left: 78.2727px; top: 661.586px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.993066);" data-canvas-width="344.0311035353532">Center of Microorganism Strains of Ukraine. We have produced</div><div style="left: 78.2727px; top: 676.081px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.977405);" data-canvas-width="343.4273888888888">haemagglutinating antigen using embryonated SPF fowl eggs and</div><div style="left: 78.2727px; top: 690.576px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.996321);" data-canvas-width="344.56089393939425">10-11 day incubation. A dilution of the virus was inoculated in</div><div style="left: 78.2727px; top: 705.071px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.03944);" data-canvas-width="342.9468813131315">0.1 ml volumes into the allantoic cavity and incubated at 35-37° C</div><div style="left: 78.2727px; top: 719.566px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.991698);" data-canvas-width="344.4167416666665">for 80-96 hours. For the purpose of NDV inactivation, we used</div><div style="left: 78.2727px; top: 734.061px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.977812);" data-canvas-width="341.8269290404039">aminoethyleneimine at the final concentration of 0.1%. Positive serum</div><div style="left: 78.2727px; top: 748.556px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.976464);" data-canvas-width="341.7135785353534">was prepared by immunizing 60-day-old chickens with live virus once</div><div style="left: 78.2727px; top: 763.051px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.01376);" data-canvas-width="342.6524164141413">and by inactivated virus twice with an interval of 2 weeks. Negative</div><div style="left: 78.2727px; top: 777.546px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.985398);" data-canvas-width="341.81830454545417">serum was obtained from healthy birds that did not contain antibodies</div><div style="left: 78.2727px; top: 792.041px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.976421);" data-canvas-width="343.8955757575757">to NDV. The investigated blood sera were inactivated by heating</div><div style="left: 78.2727px; top: 806.536px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.978858);" data-canvas-width="341.5398565656565">(56 C/30 minutes). Samples of 1% suspension of chicken erythrocytes</div><div style="left: 78.2727px; top: 821.031px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00197);" data-canvas-width="313.0814873737372">in phosphate buffered saline (pH 7.0-7.1) were used in HI tests.</div><div style="left: 78.2727px; top: 848.614px; font-size: 12.3207px; font-family: sans-serif; transform: scaleX(1.08542);" data-canvas-width="44.50239393939395">Results</div><div style="left: 91.3182px; top: 861.617px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.977258);" data-canvas-width="330.6877777777779">The specific haemagglutination activity of the obtained antigen</div><div style="left: 78.2727px; top: 876.112px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00708);" data-canvas-width="112.08763257575757">amounted to 10-11 log</div><div style="left: 190.36px; top: 884.569px; font-size: 7.39242px; font-family: serif; transform: scaleX(0.998976);" data-canvas-width="5.5443181818181815">2.</div><div style="left: 195.904px; top: 876.112px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00369);" data-canvas-width="224.69273484848483">The test was performed using the 4HA units</div><div style="left: 78.2727px; top: 890.607px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.995112);" data-canvas-width="342.13617878787875">of the antigen. Positive control serum activity was in the range of 7-9</div><div style="left: 78.2727px; top: 905.102px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00186);" data-canvas-width="15.745863636363634">log</div><div style="left: 94.0166px; top: 913.559px; font-size: 7.39242px; font-family: serif;">2</div><div style="left: 97.7128px; top: 905.102px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00081);" data-canvas-width="312.9706010101009">. Negative control serum did not give results of more than 2 log</div><div style="left: 410.678px; top: 913.559px; font-size: 7.39242px; font-family: serif;">2</div><div style="left: 414.374px; top: 905.102px; font-size: 12.3207px; font-family: serif;">.</div><div style="left: 91.3182px; top: 919.597px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.957454);" data-canvas-width="330.5768914141412">The estimation of the quality indexes of the components of the</div><div style="left: 78.2727px; top: 934.092px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.979954);" data-canvas-width="341.57928282828277">diagnostic test-kit was performed using harmonized methods. In order</div><div style="left: 78.2727px; top: 948.586px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.984919);" data-canvas-width="341.6519750000001">to examine sensitivity and specificity of HI test kits, antigens and sera</div><div style="left: 78.2727px; top: 963.081px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.02082);" data-canvas-width="342.9111512626263">from commercial diagnostic kits were used. Also, certified negative</div><div style="left: 78.2727px; top: 977.576px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00947);" data-canvas-width="342.56493939393937">control serum and samples of International Standard sera were used,</div><div style="left: 78.2727px; top: 992.071px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.989056);" data-canvas-width="343.79577803030327">which were obtained from reference laboratories, namely against</div><div style="left: 78.2727px; top: 1006.57px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.04319);" data-canvas-width="343.2080803030303">the following pathogens: Avian Influenza A (H5), Avian Influenza</div><div style="left: 78.2727px; top: 1021.06px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.966444);" data-canvas-width="343.66764267676757">A (H7), Egg Drop Syndrome’76 Virus, Paramyxoviruses of 2 and</div><div style="left: 78.2727px; top: 1035.56px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.001);" data-canvas-width="344.0619053030303">3 serotypes, Reovirus, Avian Infectious Laryngotracheitis, Avian</div><div style="left: 78.2727px; top: 1050.05px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00855);" data-canvas-width="330.2319116161615">Infectious Bronchitis Virus, Mycoplasma gallisepticum, and NDV.</div><div style="left: 456.591px; top: 290.516px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.969237);" data-canvas-width="331.01058030303017">In order to ensure a high degree of specificity for the antigen,</div><div style="left: 443.545px; top: 305.011px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.01173);" data-canvas-width="339.1976901515151">special attention was given to the selection of a stabilizer for freeze-</div><div style="left: 443.545px; top: 319.506px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00126);" data-canvas-width="154.6618358585859">drying (the subject of a patent).</div><div style="left: 456.591px; top: 334.001px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.963417);" data-canvas-width="330.6446553030302">Comparison between the national diagnostic test kit for HI and</div><div style="left: 443.545px; top: 348.496px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.03081);" data-canvas-width="345.30383257575755">commercial ELISA kit (IDEXX) in the evaluation of humoral</div><div style="left: 443.545px; top: 362.991px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.982653);" data-canvas-width="343.94485858585887">immune response to ND in vaccinated chickens was investigated</div><div style="left: 443.545px; top: 377.486px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.01815);" data-canvas-width="342.82737045454553">by examining of serum samples (n=152). Statistical analysis of data</div><div style="left: 443.545px; top: 391.98px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.982288);" data-canvas-width="341.75423686868675">showed that the correlation coefficient for the results of both tests was</div><div style="left: 443.545px; top: 406.475px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.990892);" data-canvas-width="342.02282828282824">0.92. The relative sensitivity of HI test kit was 93.5% and the relative</div><div style="left: 443.545px; top: 420.97px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00867);" data-canvas-width="96.49577777777777">specificity - 91.5%.</div><div style="left: 456.591px; top: 435.465px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.02263);" data-canvas-width="329.91773358585846">The developed test kit was successfully used for the examination</div><div style="left: 443.545px; top: 449.96px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.998681);" data-canvas-width="341.9981868686869">of field samples. We developed regulatory documents, completed the</div><div style="left: 443.545px; top: 464.455px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.988346);" data-canvas-width="341.8195366161617">procedure of validation and registration in Ukraine of the commercial</div><div style="left: 443.545px; top: 478.95px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00779);" data-canvas-width="245.6995404040405">HI test kit for the detection of antibodies to NDV.</div><div style="left: 443.545px; top: 506.533px; font-size: 12.3207px; font-family: sans-serif; transform: scaleX(1.10408);" data-canvas-width="73.93656313131314">Conclusions</div><div style="left: 456.591px; top: 519.536px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.03977);" data-canvas-width="330.1284176767676">The use of the national standardized diagnostic test kit based on</div><div style="left: 443.545px; top: 534.031px; font-size: 12.3207px; font-family: serif; transform: scaleX(0.97303);" data-canvas-width="339.2629898989899">HI for detection of antibodies to NDV allows assessing the post-</div><div style="left: 443.545px; top: 548.526px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.04125);" data-canvas-width="343.38796262626255">vaccination antibodies level that helps to maintain the disease-free</div><div style="left: 443.545px; top: 563.021px; font-size: 12.3207px; font-family: serif; transform: scaleX(1.00134);" data-canvas-width="290.18961363636356">status of the Ukrainian poultry industry with regard to ND.</div>}, number={1}, journal={Online Journal of Public Health Informatics}, author={Blotska, Oksana}, year={2017}, month={May} }