Utilization of RT-PCR and Restriction Enzyme Analysis in Detection and Differentiation of Pigeon Paramyxovirus-1 and Newcastle Disease Virus in Pigeons
Hamouda, E., Mohamed, N., Eid, A., Ismail, A., El-Sisi, M., Hassanin, O., Abou-Hashem, N. (2017). Utilization of RT-PCR and Restriction Enzyme Analysis in Detection and Differentiation of Pigeon Paramyxovirus-1 and Newcastle Disease Virus in Pigeons. EKB Journal Management System, 45(Supplementary 1), 37-47. doi: 10.21608/zvjz.2017.29239
Esraa E. Hamouda; Nagi A.A. Mohamed; Amal A M Eid; Abdelshakour N.A. Ismail; Mohamed A. El-Sisi; Ola A.A. Hassanin; Naser A. Abou-Hashem. "Utilization of RT-PCR and Restriction Enzyme Analysis in Detection and Differentiation of Pigeon Paramyxovirus-1 and Newcastle Disease Virus in Pigeons". EKB Journal Management System, 45, Supplementary 1, 2017, 37-47. doi: 10.21608/zvjz.2017.29239
Hamouda, E., Mohamed, N., Eid, A., Ismail, A., El-Sisi, M., Hassanin, O., Abou-Hashem, N. (2017). 'Utilization of RT-PCR and Restriction Enzyme Analysis in Detection and Differentiation of Pigeon Paramyxovirus-1 and Newcastle Disease Virus in Pigeons', EKB Journal Management System, 45(Supplementary 1), pp. 37-47. doi: 10.21608/zvjz.2017.29239
Hamouda, E., Mohamed, N., Eid, A., Ismail, A., El-Sisi, M., Hassanin, O., Abou-Hashem, N. Utilization of RT-PCR and Restriction Enzyme Analysis in Detection and Differentiation of Pigeon Paramyxovirus-1 and Newcastle Disease Virus in Pigeons. EKB Journal Management System, 2017; 45(Supplementary 1): 37-47. doi: 10.21608/zvjz.2017.29239

Utilization of RT-PCR and Restriction Enzyme Analysis in Detection and Differentiation of Pigeon Paramyxovirus-1 and Newcastle Disease Virus in Pigeons

Zagazig Veterinary Journal
Article 4, Volume 45, Supplementary 1, October 2017, Page 37-47  XML PDF (698.87 K)
Document Type: Original Article
DOI: 10.21608/zvjz.2017.29239
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Authors
Esraa E. Hamouda1; Nagi A.A. Mohamed2; Amal A M Eid email orcid 3; Abdelshakour N.A. Ismail1; Mohamed A. El-Sisi1; Ola A.A. Hassanin1; Naser A. Abou-Hashem1
1Avian and Rabbit Medicine Department, Faculty of Veterinary Medicine, Zagazig University, 45511, Egypt
2Veterinary Hospital, Faculty of Veterinary Medicine, Zagazig University, 45511, Egypt
3Avian and Rabbit Diseases, Faculty of Veterinary Medicine, Zagazig University, Sharkia, Egypt
Abstract
Fifty-one pigeon houses from both commercial lofts and backyard of different ages and breeds were investigated for incrimination of pigeon paramyxovirus-1 (PPMV-1) and/ or Newcastle Disease virus (NDV) during 2013-2015. The results of clinical examination revealed nervous signs (wing paralysis, head tremors, torticollis, opisthotonos and leg paralysis), greenish diarrhea, and respiratory signs, with variable mortalities. The gross lesions included congestion and hemorrhage in brain; nephrosis and/ or nephritis in kidneys, ulcer in intestine and mucoid enteritis. Virus isolation via chicken embryo inoculation revealed embryo congestion and hemagglutination (HA) activity of harvested allantoic fluids were detected 41 samples (80.39%) with HA titers ranging from 26- 210. The hemagglutination inhibition (HI) results of the same isolates against PPMV-1 and NDV hyper immune sera which prepared in rabbits (1:5dilution) varied from 23 – 210   in 38 virus isolates. The allantoic fluids of 41 haemaglutinating virus isolates were subjected to Reverse transcriptase polymeraze chain reaction (RT-PCR), Thirty-eight isolates evidenced successful products for partial amplification of fusion protein gene of NDV and PPMV-1 at 356 bp using specific primers.  Restriction fragment length polymorphism (RFLP) using   a set of three enzymes; HhaI, MspI and MboI. revealed that 9 strains (mesogenic / lentogenic PPMV-1), 27(mesogenic / lentogenic NDV), one velogenic strain for PPMV-1 and the other was NDV. MDT value in ECE revealed 72 ± 0.0 and 62.4±5.89 indicating that both isolates could be mesogenic. It could be concluded that both NDV and PPMV-1 are co- circulating among pigeons and causing economical losses. Rt-PCR followed by RFLP can be useful rapid tools for detection and identification of both viruses. Beside, in the view of the failure of adopted   prophylactic vaccination, both LaSota and Inactivated PPMV-1 vaccines must be included to overcome the impact of aforementioned disease problems in pigeons
Keywords
PPMV-1; NDV; Pigeons; PCR; differentiation; RFLP
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