scholarly journals Pathotyping of Newcastle disease virus: A novel single BsaHI digestion method of detection and differentiation of avirulent strains (lentogenic and mesogenic vaccine strains) from virulent virus

2021 ◽  
Author(s):  
Perumal Arumugam Desingu ◽  
Shambhu Dayal Singh ◽  
Kuldeep Dhama ◽  
Obli Rajendran Vinodhkumar ◽  
K Nagarajan ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Rt Pcr ◽  
Specific Primer ◽  
Digestion Method ◽  
F Gene ◽  
Virulent Strains ◽  
Virulent Virus ◽  
Short Space

We provide a novel single restriction enzyme (RE) (BsaHI) digestion approach for detecting distinct pathotypes of the Newcastle disease virus (NDV). After scanning 4000 F gene nucleotide sequences in the NCBI database, a single RE (BsaHI) digesting site was discovered in the cleavage site. APMV-I "F gene" Class II specific primer-based reverse transcriptase PCR (RT-PCR) was utilized to amplify a 535 bp fragment, which was then digested with a single RE (BsaHI) for pathotyping avian NDV field isolates and pigeon paramyxovirus-1 isolates. The avirulent (lentogenic and mesogenic strains) produce 189 and 346 bp fragments, respectively, but the result in velogenic strains remains undigested with 535 bp fragments. In addition, 45 field NDV isolates and 8 vaccine strains were used to confirm the approach. The sequence-based analysis also agrees with the data obtained utilizing the single RE (BsaHI) digestion approach. The proposed technique had the potential to distinguish between avirulent and virulent strains in a short space of time, making it valuable in NDV surveillance and monitoring research.

scholarly journals Pathotyping of Newcastle Disease Virus: a Novel Single BsaHI Digestion Method of Detection and Differentiation of Avirulent Strains (Lentogenic and Mesogenic Vaccine Strains) from Virulent Virus

2021 ◽  
Author(s):  
Perumal Arumugam Desingu ◽  
Shambhu Dayal Singh ◽  
Kuldeep Dhama ◽  
Obli Rajendran Vinodhkumar ◽  
K. Nagarajan ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Vaccine Strain ◽  
Newcastle Disease ◽  
Wild Birds ◽  
Digestion Method ◽  
Virulent Virus

The extensive use of the NDV vaccine strain and the existence of avirulent NDV strains in wild birds makes it difficult to diagnose Newcastle Disease virus (NDV). The intracerebral pathogenicity index (ICPI) and/or sequencing-based identification, which are required to determine virulent NDV, are time-consuming, costly, difficult, and cruel techniques.

scholarly journals Newcastle Disease Virus Detection from Chicken Organ Samples Using Reverse Transcriptase Polymerase Chain Reaction

2017 ◽  
Vol 35 (1) ◽  
pp. 127
Author(s):  
Lehgarubini Shanmuganathan ◽  
Dito Anggoro ◽  
Michael Haryadi Wibowo
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Rna Extraction ◽  
Rt Pcr ◽  
Chain Reaction ◽  
F Gene ◽  
Polymerase Chain

Newcastle disease (ND) is a systemic, viral respiratory disease that is acute and easily transmitted which affects various types of poultry, especially chickens. Diagnosis of ND which generally involves virus isolation and subsequent identification with serological assays has limitations that needs more time. This research was aimed to detect Newcastle Disease virus (NDV) in chickens suspected with ND using the Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) technique. Nine chicken organ samples such as lien, trachea, and lungs were collected from chicken farms diagnosed with ND. The organ samples were processed and the targeted viral RNA was extracted using the RNA extraction kit. Genome amplification was performed with RT-PCR using specificprimers to target the F gene. Amplification results produced an amplicon product of 565 base pairs (bp). PCR product samples were then visualised using agar gel electrophoresis and viewed using the unified gel documentation system. Amplification results show nine samples positive for the DNA bands corresponding to the targeted band of the NDV F gene fragment. The results of this research confirm that the RT-PCR method is applicable for NDV detection from chicken organ samples.

scholarly journals Complete Genome Sequences of Two Newcastle Disease Virus Strains Isolated from a Wild Duck and a Pigeon in Russia

2016 ◽  
Vol 4 (6) ◽  
Author(s):  
Marsel R. Kabilov ◽  
Tatyana Y. Alikina ◽  
Kseniya S. Yurchenko ◽  
Alexandra V. Glushchenko ◽  
Konstantin V. Gunbin ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Complete Genome ◽  
Comparative Sequence Analysis ◽  
Genome Sequences ◽  
Wild Duck ◽  
F Gene ◽  
Virus Strains

Here, we report the complete genome sequences of two Newcastle disease virus (NDV) isolates, Adygea/duck/12/2008, from a wild duck in Russia, and Altai/pigeon/777/2010, from a pigeon in Russia. Based on comparative sequence analysis of the F gene, these strains were classified as NDV class II, genotypes VIId and VIb/2, respectively.

scholarly journals Newcastle Disease Viruses Causing Recent Outbreaks Worldwide Show Unexpectedly High Genetic Similarity to Historical Virulent Isolates from the 1940s

2016 ◽  
Vol 54 (5) ◽  
pp. 1228-1235 ◽  
Author(s):  
Kiril M. Dimitrov ◽  
Dong-Hun Lee ◽  
Dawn Williams-Coplin ◽  
Timothy L. Olivier ◽  
Patti J. Miller ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Rna Virus ◽  
Phylogenetic Analyses ◽  
Genetic Distances ◽  
Wild Birds ◽  
Rates Of Change ◽  
Virulent Strains ◽  
Experimental Challenge

Virulent strains of Newcastle disease virus (NDV) cause Newcastle disease (ND), a devastating disease of poultry and wild birds. Phylogenetic analyses clearly distinguish historical isolates (obtained prior to 1960) from currently circulating viruses of class II genotypes V, VI, VII, and XII through XVIII. Here, partial and complete genomic sequences of recent virulent isolates of genotypes II and IX from China, Egypt, and India were found to be nearly identical to those of historical viruses isolated in the 1940s. Phylogenetic analysis, nucleotide distances, and rates of change demonstrate that these recent isolates have not evolved significantly from the most closely related ancestors from the 1940s. The low rates of change for these virulent viruses (7.05 × 10−5and 2.05 × 10−5per year, respectively) and the minimal genetic distances existing between these and historical viruses (0.3 to 1.2%) of the same genotypes indicate an unnatural origin. As with any other RNA virus, Newcastle disease virus is expected to evolve naturally; thus, these findings suggest that some recent field isolates should be excluded from evolutionary studies. Furthermore, phylogenetic analyses show that these recent virulent isolates are more closely related to virulent strains isolated during the 1940s, which have been and continue to be used in laboratory and experimental challenge studies. Since the preservation of viable viruses in the environment for over 6 decades is highly unlikely, it is possible that the source of some of the recent virulent viruses isolated from poultry and wild birds might be laboratory viruses.

Isolation and Molecular Characterization of Newcastle Disease Virus in Layers

2021 ◽  
Author(s):  
Smita Bordoloi ◽  
Anju Nayak ◽  
A.P. Singh ◽  
R.V. Singh ◽  
Kajal Jadav ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Virulent Strain ◽  
Indian Subcontinent ◽  
Economic Losses ◽  
Tissue Samples ◽  
F Gene ◽  
Virulent Newcastle Disease Virus

Background: Newcastle disease (ND) in spite of the availability of vaccines remains a constant threat to poultry producers worldwide. It is prevalent in Indian subcontinent and leads to economic losses. The present study was aimed with isolate and identify virulent Newcastle disease virus (NDV) in layer poultry from field outbreaks.Methods: Total 47 samples consisting of nasal (05), oropharyngeal (13) and cloacal swabs (11) and tissue samples consisting of trachea (07), lungs (06), larynx (05) were collected from layer birds. For isolation of NDV swab and tissue samples were inoculated in 9-11 days old embryonated eggs via allantoic cavity route. After preparing the viral inoculum, 47 suspected samples (29 swab and 18 tissue samples) were inoculated in 141 embryonated eggs to isolate the virus.Result: Out of 47 samples 10 (21.27%) samples were positive for HA activity. All the 10 isolates showing HA activity subjected to Reverse-Transcriptase PCR of F gene and 6 were found positive in RT-PCR for F1 gene. The PCR amplified product showed amplicon at 356 bp and 254 bp positive for F1 and F2 gene, respectively. On basis of F gene, 06 (50%) isolates were considered as virulent Newcastle Disease Virus. One isolate sequence was submitted at NCBI with accession MT890653 On phylogenetic analysis MT890653 designated as Class II/ genotype II/ virulent strain and had the motif 112R-R-R-K-R-F117 at the cleavage site of the fusion protein.

Evaluation of RT-PCR for Rapid Detection of Sudanese Isolates and Vaccine Strains of Newcastle Disease Virus

2005 ◽  
Vol 8 (3) ◽  
pp. 418-420
Author(s):  
Mohamed A.M. Yousof . ◽  
I.E. Aradaib . ◽  
K.M.S. Khairalla . ◽  
M.A. Abdalla . ◽  
A.R.E. Karrar . ◽  
...  
Keyword(s):  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Rapid Detection ◽  
Rt Pcr

scholarly journals First Genome Sequence of Newcastle Disease Virus of Genotype VIIi from Jordan

2018 ◽  
Vol 7 (23) ◽  
Author(s):  
Mustafa Ababneh ◽  
Helena L. Ferreira ◽  
Mohammad Khalifeh ◽  
David L. Suarez ◽  
Claudio L. Afonso
Keyword(s):  
Reverse Transcriptase ◽  
Newcastle Disease Virus ◽  
Disease Virus ◽  
Genome Sequence ◽  
Illumina Sequencing ◽  
Newcastle Disease ◽  
Rt Pcr ◽  
Total Rna ◽  
Reverse Transcriptase Pcr ◽  
Chicken Spleen

Newcastle disease virus (NDV) was detected by reverse transcriptase PCR (RT-PCR) from total RNA isolated from a chicken spleen of a backyard flock in Jordan. The complete coding genome sequence of NDV/chicken/Jordan/J11-spleen/2018 was obtained with MiSeq (Illumina) sequencing.

scholarly journals Real-Time Reverse Transcription-Polymerase Chain Reaction Detection of Newcastle Disease Virus Using Light upon Extension Fluorogenic Primers

2007 ◽  
Vol 19 (4) ◽  
pp. 400-404 ◽  
Author(s):  
Márta Antal ◽  
Tibor Farkas ◽  
Péter Germán ◽  
Sándor Belák ◽  
István Kiss
Keyword(s):  
Newcastle Disease Virus ◽  
Real Time ◽  
Disease Virus ◽  
Newcastle Disease ◽  
Plasmid Copy Number ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Plasmid Copy ◽  
Infectious Dose ◽  
Efficient Detection

A real-time reverse transcriptase (RT)-PCR assay, applying light upon extension (LUX) fluorogenic primers, was developed for rapid and efficient detection of Newcastle disease virus (NDV). The method, which targets the fusion (F) protein gene of the viral genome, gave positive signal with all NDV isolates tested (32/32), while negative results were obtained with heterologous pathogens (35/35), including 13 avian influenza virus isolates. The detection limit of the assay was approximately 10+1.2 egg infectious dose (EID)50/0.2 ml and 10+2.2 EID50/0.2 ml for virus suspensions and spiked chicken fecal samples, respectively. As expressed in plasmid copy number, the procedure has a sensitivity of approximately 20 copies of the plasmid harboring the target gene. Due to its high specificity, sensitivity, and relative simplicity, the LUX RT-PCR assay provides a novel, rapid, and practical tool for the detection of NDV.

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