Molecular Characterization of Velogenic Newcastle Disease Virus (Sub-Genotype VII.1.1) from Wild Birds, with Assessment of Its Pathogenicity in Susceptible Chickens

Newcastle disease (ND) is considered to be one of the most economically significant avian viral diseases. It has a worldwide distribution and a continuous diversity of genotypes. Despite its limited zoonotic potential, Newcastle disease virus (NDV) outbreaks in Egypt occur frequently and result in serious economic losses in the poultry industry. In this study, we investigated and characterized NDV in wild cattle egrets and house sparrows. Fifty cattle egrets and fifty house sparrows were collected from the vicinity of chicken farms in Kafrelsheikh Governorate, Egypt, which has a history of NDV infection. Lung, spleen, and brain tissue samples were pooled from each bird and screened for NDV by real-time reverse transcriptase polymerase chain reaction (RRT-PCR) and reverse transcriptase polymerase chain reaction (RT-PCR) to amplify the 370 bp NDV F gene fragment. NDV was detected by RRT-PCR in 22 of 50 (44%) cattle egrets and 13 of 50 (26%) house sparrows, while the conventional RT-PCR detected NDV in 18 of 50 (36%) cattle egrets and 10 of 50 (20%) of house sparrows. Phylogenic analysis revealed that the NDV strains identified in the present study are closely related to other Egyptian class II, sub-genotype VII.1.1 NDV strains from GenBank, having 99.7–98.5% identity. The pathogenicity of the wild-bird-origin NDV sub-genotype VII.1.1 NDV strains were assessed by experimental inoculation of identified strains (KFS-Motobas-2, KFS-Elhamoul-1, and KFS-Elhamoul-3) in 28-day-old specific-pathogen-free (SPF) Cobb chickens. The clinical signs and post-mortem changes of velogenic NDV genotype VII (GVII) were observed in inoculated chickens 3 to 7 days post-inoculation, with 67.5–70% mortality rates. NDV was detected in all NDV-inoculated chickens by RRT-PCR and RT-PCR at 3, 7, and 10 days post-inoculation. The histopathological findings of the experimentally infected chickens showed marked pulmonary congestion and pneumonia associated with complete bronchial stenosis. The spleen showed histocytic cell proliferation with marked lymphoid depletion, while the brain had malacia and diffuse gliosis. These findings provide interesting data about the characterization of NDV in wild birds from Egypt and add to our understanding of their possible role in the transmission dynamics of the disease in Egypt. Further research is needed to explore the role of other species of wild birds in the epidemiology of this disease and to compare the strains circulating in wild birds with those found in poultry.

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A244 CHARACTERIZATION OF THE EARLY INFLAMMATORY RESPONSE IN A BABOON MODEL OF ACUTE RESPIRATORY FAILURE (ARF) BY REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION (RT-PCR)

Anesthesiology ◽

10.1097/00000542-199709001-00244 ◽

1997 ◽

Vol 87 (Supplement) ◽

pp. 244A

Author(s):

P. Germann ◽

G. Roder ◽

G. Urak ◽

G. Schlag ◽

M. Zimpfer ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Respiratory Failure ◽

Inflammatory Response ◽

Acute Respiratory Failure ◽

Rt Pcr ◽

Chain Reaction ◽

Baboon Model ◽

Early Inflammatory Response ◽

Polymerase Chain

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Detection of Newcastle disease virus of poultry by real time reverse transcription-polymerase chain reaction

Bangladesh Veterinarian ◽

10.3329/bvet.v33i1.33309 ◽

2017 ◽

Vol 33 (1) ◽

pp. 16-22

Author(s):

MM Rahman ◽

LR Barman ◽

EH Chowdhury ◽

MR Islam

Keyword(s):

Polymerase Chain Reaction ◽

Newcastle Disease Virus ◽

Disease Virus ◽

Reverse Transcription ◽

Newcastle Disease ◽

Allantoic Fluid ◽

Rt Pcr ◽

M Gene ◽

Chain Reaction ◽

Polymerase Chain

A real-time reverse transcription - polymerase chain reaction (rRT-PCR) was used for the detection of Newcastle disease virus (NDV) of poultry. A panel of seven known isolates of NDV in the form of allantoic fluid, obtained from a laboratory repository, was used for the development of the test. RNA was extracted from the allantoic fluid with a magnetic processor based automated RNA extraction system. The identity of the reference virus was first reconfirmed by a conventional RT-PCR specific for the Fusion (F) protein gene. Using these RNA, the rRT-PCR protocol was optimized with regard to the reaction mix and thermal profile using published primers and probes specific for M gene. The sensitivity of standardized rRT-PCR was compared to that of the conventional RT-PCR using serial 10-fold dilutions of the RNA of a selected sample. The thermal profile was modified from the published one; the annealing and extension steps were combined to a single step performed at 60ºC. The adopted rRT-PCR successfully amplified M gene from all the seven reference samples with a CT value ranging from 15.28 to 32.68. The rRT-PCR for M gene was 100-fold more sensitive than the conventional RT-PCR for F gene. This is the first report of the use of rRT-PCR for the detection of NDV in Bangladesh. This test will be useful for virological surveillance, particularly for screening NDV in respiratory infections.Bangl. vet. 2016. Vol. 33, No. 1, 16-22

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Evidence for airborne transmission of Norwalk-like virus (NLV) in a hotel restaurant

Epidemiology and Infection ◽

10.1017/s0950268899003805 ◽

2000 ◽

Vol 124 (3) ◽

pp. 481-487 ◽

Cited By ~ 227

Author(s):

P. J. MARKS ◽

I. B. VIPOND ◽

D. CARLISLE ◽

D. DEAKIN ◽

R. E. FEY ◽

...

Keyword(s):

Electron Microscopy ◽

Polymerase Chain Reaction ◽

Inverse Relationship ◽

Nucleotide Sequence Analysis ◽

Rt Pcr ◽

Chain Reaction ◽

Virus Particles ◽

Stool Samples ◽

Polymerase Chain

An outbreak of gastroenteritis followed a meal in a large hotel during which one of the diners vomited. The clinical features of the illness suggested Norwalk-like virus (NLV, small round structured virus) infection, and this was confirmed by electron microscopy and reverse transcriptase polymerase chain reaction (RT–PCR) of stool samples. Further characterization of the virus by nucleotide sequence analysis of the PCR amplicons revealed identical strains in all the affected individuals. The foods served at the meal could not be demonstrated to be the cause of the outbreak. Analysis of attack rates by dining table showed an inverse relationship with the distance from the person who vomited. No one eating in a separate restaurant reported illness. Transmission from person-to-person or direct contamination of food seems unlikely in this outbreak. However, the findings are consistent with airborne spread of NLV with infection by inhalation with subsequent ingestion of virus particles.

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Characterization of myxomycete communities in soil by reverse transcription polymerase chain reaction (RT-PCR)-based method

Soil Biology and Biochemistry ◽

10.1016/j.soilbio.2009.04.001 ◽

2009 ◽

Vol 41 (6) ◽

pp. 1324-1330 ◽

Cited By ~ 9

Author(s):

Akiko Kamono ◽

Jun Matsumoto ◽

Hisaya Kojima ◽

Manabu Fukui

Keyword(s):

Polymerase Chain Reaction ◽

Reverse Transcription ◽

Rt Pcr ◽

Chain Reaction ◽

Polymerase Chain

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

Jurnal Sain Veteriner ◽

10.22146/jsv.29300 ◽

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.

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Production of Newcastle Disease Polyclonal Antibody as the Alternative of Immunohistochemistry Primary Antibody against Newcastle Disease in Poultry

World s Veterinary Journal ◽

10.54203/scil.2021.wvj24 ◽

2021 ◽

Vol 11 (2) ◽

pp. 187-192

Author(s):

Muhammad Kholish Naf’an ◽

Kurniasih Kurniasih ◽

Tri Untari ◽

Yos Adi Prakoso

Keyword(s):

Polymerase Chain Reaction ◽

New Zealand ◽

Polyclonal Antibody ◽

Newcastle Disease ◽

Primary Antibody ◽

Rt Pcr ◽

Chain Reaction ◽

New Zealand White Rabbits ◽

Polymerase Chain ◽

Similar Accuracy

Newcastle disease (ND) is the most pathogenic viral infection in poultry. Furthermore, the availability of laboratories that support the molecular diagnosis of ND is still limited in Indonesia. The present study aimed to produce ND polyclonal antibody as the alternative of immunohistochemistry primary antibody against ND in poultry. Two adult male New Zealand White rabbits weighed 2.5 kg were vaccinated seven days after the adaptation using intraperitoneal injection of the ND live vaccine at multilevel doses weekly. The serum was collected inactivated, and purified in the sixth week. A total number of 31 chicken samples were collected and their samples of brain, lung, spleen, and intestine were tested using immunohistochemistry and Reverse Transcription Polymerase Chain reaction (RT-PCR). The result showed that 19/31 (61%) were positive against immunohistochemistry and RT-PCR and a total of 12/31 (39%) were negative. Based on the obtained results, immunohistochemistry using ND polyclonal antibody had a similar accuracy with RT-PCR. It can be concluded that ND polyclonal antibody produced by vaccination in the rabbit could be used as the alternative immunohistochemistry primary antibody for diagnosing ND in poultry.

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Molecular cloning and pharmacological characterization of the rat sigma1 receptor11Abbreviations: DTG, N,N′-di(o-tolyl-guanidine); HA, hemagglutinin; RT-PCR, reverse transcriptase-polymerase chain reaction; RACE, rapid amplification of cDNA ends; and CHO, Chinese hamster ovary.

Biochemical Pharmacology ◽

10.1016/s0006-2952(01)00666-9 ◽

2001 ◽

Vol 62 (3) ◽

pp. 349-355 ◽

Cited By ~ 78

Author(s):

Jianfeng Mei ◽

Gavril W. Pasternak

Keyword(s):

Polymerase Chain Reaction ◽

Molecular Cloning ◽

Chinese Hamster Ovary ◽

Chinese Hamster ◽

Rt Pcr ◽

Chain Reaction ◽

Pharmacological Characterization ◽

Polymerase Chain ◽

Rapid Amplification

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Penentuan Patogenesitas Molekuler Virus Newcastle Disease yang Diisolasi dari Ayam Komersial Tahun 2013-2016

Acta Veterinaria Indonesiana ◽

10.29244/avi.5.2.105-119 ◽

2018 ◽

Vol 5 (2) ◽

pp. 105-119

Author(s):

Sarwo Edy Wibowo ◽

Michael Haryadi Wibowo ◽

Bambang Sutrisno

Keyword(s):

Polymerase Chain Reaction ◽

Avian Influenza ◽

Newcastle Disease ◽

Infectious Bursal Disease ◽

Rt Pcr ◽

Chain Reaction ◽

Infectious Bronchitis ◽

Bursal Disease ◽

Infectious Laryngotracheitis ◽

Polymerase Chain

Newcastle Disease (ND) atau yang dikenal dengan “Tetelo” masih menjadi masalah di peternakan unggas komersil, meskipun telah dilakukan vaksinasi ND secara rutin, namun wabah ND masih tetap terjadi. Penelitian ini bertujuan untuk mengetahui patogenesitas molekuler dan genotipe virus ND berdasarkan analisis sekuen fragmen gen F, serta melihat hubungan kekerabatan antara isolat dalam penelitian dengan isolat ND di Indonesia sebelumnya serta strain vaksin pada peternakan ayam yang menerapkan vaksinasi ND secara berkala. Penelitian ini menggunakan primer yang didesain dengan konsensus fragmen gen F dari GenBank dan desain dengan aplikasi amplifX pada posisi 91-800 nt dengan panjang 710bp. Urutan basa pada primer kemudian di cek dengan BLAST primer dan di uji spesifisitas dengan beberapa virus penyakit unggas yaitu vaksin infectious bronchitis (IB) 120, virus vaksin infectious laryngotracheitis (ILT), virus vaksin avian influenza (AI), dan virus vaksin infectious bursal disease (IBD). Delapan sampel paru diperoleh dari delapan peternakan ayam komersial di Yogyakarta, Semarang, Jakarta, Magelang dan Muntilan. Tiga sampel diisolasi pada telur ayam berembrio dan diidentifikasi menggunakan uji HA dan HI. Lima sampel lain dilakukan ekstraksi secara langsung dari gerusan organ paru. Sampel di identifikasi dengan metode reverse transciptase-polymerase chain reaction (RT-PCR) untuk mendeteksi gen F menggunakan primer forward 5’ TCT CTT GAT GGC AGG CCT CTT G ‘3 dan reverse 5’ CCG CTA CCG ATT AAT GAG CTG AGT’3 dengan panjang produk 710 bp. Hasil penelitian menunjukkan bahwa seluruh sampel positif virus ND. Analisis hasil sekuen dengan menggunakan perangkat lunak MEGA v.7 didapatkan susunan asam amino penyusun cleavage site 112RRQKR↓F117 dan 112RRRKR↓F117yang menunjukkan bahwa virus ND tersebut digolongkan strain velogenik. Berdasarkan analisis pohon filogenetik isolat ND-Layer/GK-SR1/2013, ND-Lay/Pullet-80/ 27/16 (N), ND-Bro/Lingga 2L/24/2 (N), dan ND-Lay/Smg-P/2015 merupakan genotip VIIi, sedangkan 3 isolat ND yaitu ND-Bro/Yog-P/2015, JKT/P1/2016 (Jakarta), dan JKT/P2/2016 (Jakarta) merupakan ND genotip VIIh. Jarak genetik isolat yang diteliti dengan virus ND Indonesia yang pernah dilaporkan sebelumnya pada fragmen gen F posisi 91-798 berkisar 0,4 – 9,6 % dengan tingkat homologi mencapai 90,4 – 99,5%.

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