Newcastle disease (ND) is a highly contagious and economically important disease in the poultry industry caused by avian avulavirus-1, historically known as Newcastle disease virus (NDV). Control of ND primarily relies on prophylactic vaccination of flocks, and many vaccines are available on the market, both conventional and more recently introduced new generation recombinant types. To assess the protection level achieved by vaccination ELISA tests are typically used, they also are to track an infection with field strains in non-vaccinated flocks. Special modifications of ELISA can be used as a screening tool to detect infection in flocks vaccinated with new generation vaccines. In this study, we have developed an ELISA test for the detection of antibodies against the nucleoprotein (NP) of NDV and for differentiation of chickens vaccinated with commercial and prototype in-house recombinant vector vaccines from those infected with field NDV strains. The NP gene of LaSota NDV strain expressed in a baculovirus vector was used as a coating antigen in the ELISA. The developed test was optimized, validated and compared to other serological tests. The sensitivity, specificity and accuracy of recombinant NP protein-based ELISA were respectively 96.1%, 96.3%, and 96.2%. Inter-rater (kappa) agreement between the NP-ELISA and the gold standard HI test was calculated to be 0.995. In our comparisons, commercially available ELISA tests revealed different specificities ranging from 95.5–100% and sensitivities at variance, ranging from 90.1 to 99.0%. A high level of maternally derived antibodies was measured in the serum of 1-day-old broilers in the NP-ELISA assay. These antibodies had disappeared and were undetected at 3, 5 and 6 weeks post-vaccination but birds became positive again at 2 weeks after control infection with a velogenic NDV strain. In SPF chickens, antibodies against NP protein were detected only after a challenge. The recombinant NP protein-based ELISA test is sensitive, specific and accurate when compared to the gold standard HI test and commercially available kits. Moreover, the method could be also used for the differentiation between vaccinated and infected birds.
Field surveys were undertaken in 1997–1999 across five ecological zones in Nigeria to collect isolates of Maize streak virus (MSV), genus Mastrevirus. Apart from maize (Zea mays L.), 15 other grass species were found with MSV symptoms in Nigeria. These hosts showed two types of symptoms viz: mild (with or without mottle) or severe (typical symptoms in maize). When Cicadulina storeyi China was used to attempt transmission of these isolates of MSV to seedlings a susceptible maize hybrid CML 254 X CML 247, six isolates were not transmissible to maize. Seven isolates that were transmissible to maize produced mild symptoms. The viral agents causing typical or severe streak symptoms in Axonopus compressus (Sw.) P. Beauv., Brachiaria distichophylla (Trn.) Stapf, Dactyloctenium aegyptium (Linn.) P. Beauv. and Setaria barbata (Lam.) Kunth produced symptoms that were typical of MSV in farmers fields, when transmitted to maize. Out of 33 plant species that seedlings were challenged with MSV, only eight proved susceptible. Four of them showed mild symptoms while the other four showed severe symptoms of MSV. Only three isolates collected during the surveys did not react with a MSV polyclonal antiserum produced in mice in Double Antibody Sandwich-Enzyme-Linked Immunosorbent Assay (DAS-ELISA). These isolates were found in Andropogon gayanus Kunth (from Kaduna), Thelepogon elegans Roth ex Toem & Schult (from Kadawa) and Rottboellia cochinchinensis (Lour.) Clayton (from Jos) exhibited mild streak/mottle symptoms. Specific monoclonal antibodies, raised against MSV, reacted with 12 out of 25 samples tested. The DAS-ELISA data also showed significant variation in concentration of the virus in the different plant hosts. The relationship dendogram through SDS-PAGE among eight purified virus isolates show 55–90% variation. At 0.55 coefficient of similarity, the dendogram divided the samples into two groups while at 0.9 coefficient of similarity, the 8 isolates were identified as distinct genetic entities.
In the spring of 2019, many plants, mainly winter wheat, were observed to have dwarfism and leaf yellowing symptoms. These plants from several regions of Poland were collected and sent to the Plant Disease Clinic of the Institute of Plant Protection – National Research Institute in Poznań to test for the presence of viral diseases. Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) results showed numerous cases of Wheat dwarf virus (WDV) and a few cases of plant infections caused by Barley yellow dwarf viruses (BYDVs). WDV was detected in 163 out of 236 tested winter wheat plants (69.1%), in 10 out of 27 tested winter barley plants (37%) and in 6 out of 7 triticale plants (85.7%) while BYDVs were found, respectively, in 9.7% (23 out of 236) and in 18.5% (5 out of 27) of tested winter forms of wheat and barley plants. Infected plants came mainly from the regions of Lower Silesia and Greater Poland. Furthermore, individual cases of infections were also confirmed in the following districts: Lubusz, Opole, Silesia, Kuyavia-Pomerania and Warmia-Masuria. Results of Duplex-immunocapture-polymerase chain reaction (Duplex-IC-PCR) indicated the dominance of WDV-W form in wheat and WDV-B form in barley plants. Moreover, results of reverse transcription – polymerase chain reaction (RT-PCR) connected with restriction fragment length polymorphism (RFLP) analysis, performed for 17 BYDVs samples, revealed 8 BYDV-PAS, 4 BYDV-MAV and 2 BYDVPAV as well as the presence of two mixed infections of BYDV-MAV/-PAS and one case of BYDV-MAV/-PAV. Next, RT-PCR reactions confirmed single BYDV-GAV infection and the common presence of BYDV-SGV. To the best of our knowledge, in 2020 the viruses were not a big threat to cereal crops in Poland.