Open Access
R esearch
(Published
online: 12-01-2016)
5.
Analysis of viral protein-2 encoding gene
of avian encephalomyelitis virus from field specimens in Central
Java region, Indonesia -
Aris Haryanto, Ratna Ermawati, Vera Wati, Sri Handayani
Irianingsih and Nastiti Wijayanti
Veterinary World, 9(1): 25-31
doi:
10.14202/vetworld.2016.25-31
Aris Haryanto:
Department of Biochemistry, Faculty of Veterinary Medicine,
Universitas Gadjah Mada, Yogyakarta, Indonesia; arisharyanto@yahoo.com
Ratna Ermawati:
Department of Biochemistry, Faculty of Veterinary Medicine,
Universitas Gadjah Mada, Yogyakarta, Indonesia; ratna.ermawati@yahoo.com
Vera Wati:
Division of Biotechnology, Animal Disease Investigation Center
Wates, Daerah Istimewa Yogyakarta Province,
Indonesia; verawatisutopo@gmail.com
Sri Handayani Irianingsih:
Division of Virology, Animal Disease Investigation Center Wates,
Daerah Istimewa Yogyakarta Province,
Indonesia; handa_nie@yahoo.com
Nastiti Wijayanti:
Department of Animal Physiology, Faculty of Biology, Universitas
Gadjah Mada, Yogyakarta, Indonesia; nastitiw@yahoo.com
Received: 05-08-2015, Revised: 25-11-2015, Accepted: 02-12-2015,
Published online: 12-01-2016
Corresponding author:
Aris Haryanto, e-mail: arisharyanto@yahoo.com,
arisharyanto@ugm.ac.id
Citation:
Haryanto A, Ermawati R, Wati V, Irianingsih SH, Wijayanti N
(2016) Analysis of viral protein-2 encoding gene of avian
encephalomyelitis virus from field specimens in Central Java
region, Indonesia,
Veterinary World 9(1):
25-31.
Abstract
Aim:
Avian encephalomyelitis (AE) is a viral disease which can infect
various types of poultry, especially chicken. In Indonesia, the
incidence of AE infection in chicken has been reported since
2009, the AE incidence tends to increase from year to year. The
objective of this study was to analyze viral protein 2 (VP-2)
encoding gene of AE virus (AEV) from various species of birds in
field specimen by reverse transcription polymerase chain
reaction (RT-PCR) amplification using specific nucleotides
primer for confirmation of AE diagnosis.
Materials and Methods:
A total of 13 AEV samples are isolated from various species of
poultry which are serologically diagnosed infected by AEV from
some areas in central Java, Indonesia. Research stage consists
of virus samples collection from field specimens, extraction of
AEV RNA, amplification of VP-2 protein encoding gene by RT-PCR,
separation of RT-PCR product by agarose gel electrophoresis, DNA
sequencing and data analysis.
Results:
Amplification products of the VP-2 encoding gene of AEV by
RT-PCR methods of various types of poultry from field specimens
showed a positive results on sample code 499/4/12 which
generated DNA fragment in the size of 619 bp. Sensitivity test
of RT-PCR amplification showed that the minimum concentration of
RNA template is 127.75 ng/μl. The multiple alignments of DNA
sequencing product indicated that positive sample with code
499/4/12 has 92% nucleotide homology compared with AEV with
accession number AV1775/07 and 85% nucleotide homology with
accession number ZCHP2/0912695 from Genbank database. Analysis
of VP-2 gene sequence showed that it found 46 nucleotides
difference between isolate 499/4/12 compared with accession
number AV1775/07 and 93 nucleotides different with accession
number ZCHP2/0912695.
Conclusions:
Analyses of the VP-2 encoding gene of AEV with RT-PCR method
from 13 samples from field specimen generated the DNA fragment
in the size of 619 bp from one sample with sample code 499/4/12.
The sensitivity rate of RT-PCR is to amplify the VP-2 gene of
AEV until 127.75 ng/μl of RNA template. Compared to Genbank
databases, isolate 499/4/12 has 85% and 92% nucleotide homology.
Keywords:
avian encephalomyelitis, reverse transcription polymerase chain
reaction, viral protein 2 gene.
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