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Yasunobu Matsumoto
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Kloning, Sikuensing dan Analisis Filogenetik Gen Nukleokapsid Protein Virus Tetelo Isolat Bali-1/07 Anak Agung Ayu Mirah Adi; I Made Kardena; Nyoman Mantik Astawa; Ketut Santhia Adhy Putra; Yoshihiro Hayashi; Yasunobu Matsumoto
Jurnal Veteriner Vol 12, No 3 (2011)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

A study was carried out on the molecular characteristics of gene encoding for nucleocapsid protein(NP) of Newcastle disease virus (NDV) Bali-1/07. A portion of the fragment gene was amplified by reversetranscriptase-polymerase chain reaction (RT-PCR). The RT-PCR product purified from the gel was treatedwith Taq polymerase and cloned into plasmid pT7blueT vector. The recombinant plasmid was tranfectedinto Nova blue singles strain of Escherichia coli-competent cells and selected using white and blue colorselection method. Good white colonies were subcultured and tested for presence of expected gene fragmentby polymerase chain reaction (PCR). The correct size PCR product was recloned and sequenced. The PCRproduct of 540 base pairs were sequenced and aligned with several cognates genes of world NDVisolates.using Cluster IW. The phylogenetic analysis was then performed using MEGA 4. Phylogeneticanalysis showed that the NP gene of NDV Bali-1/07 is closely related with virulent NDVs such asGuangxii-11/03, KBNP-4152 and Ast2755/01 with nucleotide sequence homology of 92%, 91.4% and91%respectively, whereas the nucleotide sequence homology with avirulent NDVs such as LaSota and B1were 77%.
Pelacakan Secara Imunohistokimiawi Antigen Virus pada Ayam yang Diinfeksi dengan Virus Penyakit Tetelo (IMMUNOHISTOCHEMICAL DETECTION OF VIRAL ANTIGEN IN TISSUE OF CHICKENS EXPERIMENTALLY INFECTED WITH NEWCASTLE DISEASE VIRUS) Anak Agung Ayu Mirah Adi; I Made Kardena; Nyoman Mantik Astawa; Yasunobu Matsumoto
Jurnal Veteriner Vol 13 No 3 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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In order to study the distribution of Newcastle disease virus (NDV) following infection, chickenswere experimentally infected with visceretropic velogenic NDV isolate. Monoclonal antibodies (mAbs)against the NDV LaSota vaccine strain were then produced to detect viral antigen in the infectedorgans. The mAbs were firstly tested for their specificity by enzyme linked immunosorbent assay(ELISA) using NDV and normal allantoic fluids as antigens. Eight mAbs specific against NDVwere isolated and two mAbs were used for immunodetection of NDV antigen in chicken’s tissues.By immunohistochemistry labeled streptavidin-biotin (LSAB) staining NDV–antigen was detectedin paraffin embedded tissues of NDV-infected chickens. NDV antigen was not detected in noninfected chickens. In the infected chickens, high intensity of NDV antigen was detected in thelymphoid tissues, lung and intestine. The NDV antigen with a lesser intensity was detected in thebrain, trachea, liver and myocardium. This study shows that although viscerotropic velogenicNDV isolate can infect almost all organs, the main target of infection are lung, intestine andlymphoids tissues
DETECTION OF NEWCASTLE DISEASE VIRUS BY NESTED REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION Anak Agung Ayu Mirah Adi; Nyoman Mantik Astawa; Ketut Santhia Adhy Putra; Yasunobu Matsumoto
Jurnal Veteriner Vol 9 No 3 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

A study to utilize nested reverse trancriptase-polymerase chain reaction (RT-PCR) for the detection ofNewcastle disease virus (NDV) infection was carried out. NDV isolated from an outbreak in KarangasemDistrict of Bali, Indonesia was propagated in chicken embryos and its pathogenicity was determined byinoculation in 3 week-old chickens. Organ samples were collected from infected chickens for nested-RTPCR.Out of several different pairs of primers designed for study, 3 pairs of primers (F4s-F6r/F5s-F5r,F8s-F10r/F8s-F8r and F12s-F14s/F13s-F13r), each of which for first-round/nested PCR, was able to amplifyspecific regions of NDV genome. In the fist-round PCR, the PCR product of 1500 bps was not clearly visiblein the agarose gel following electrophoresis. In nested PCR a PCR product of 500 bp was clearly visible onagarose gel following electrophoresis. The 3 pairs of primers appeared to be potential for an accurate andrapid detection NDV in the infected host.
Co-Authors Anak Agung Ayu Mirah Adi DWI SURYANTO I Made Kardena I NYOMAN MANTIK ASTAWA Ketut Santhia Adhy Putra Yoshihiro Hayashi