Analysis of viral protein-2 encoding gene of avian encephalomyelitis virus from field specimens in Central Java region, Indonesia

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.

In vitro expression of the recombinant fusion protein of Newcastle disease virus from local Indonesian isolates by using a cell-free protein expression system

“…Office International des Epizooties (OIE) categorizes ND in the A list of animal diseases, which includes infectious diseases with rapid and serious spreading and affecting public health and socio economic communities ( [OIE] Office Internationalof Epi zootics 2015). As other infectious viral disease outbreak which had been reported in Indonesia poultry industry, such as avian influenza (Njoto et al 2018), infectious bur sal disease (Parede et al 2003), and avian encephalomyeli tis (Haryanto et al 2016). An outbreak of ND can cause a devastating effect as it causes almost 100% mortality in susceptible poultry (Alexander et al 2012).…”

“…The expression vector used in this work had been prepared by Haryanto et al (2016) and Wulanjati et al (2018) by subcloning the F protein encoding gene of NDV from lo cal isolate in Kulon Progo, Indonesia (0663/04/2013) into pBT7NHis expression vector. The design of the recom binant plasmid vector is shown in Figure 1.…”

Detection by Rt-Pcr and Molecular Characterization of Tremovirus A Obtained from Clinical Cases of Avian Encephalomyelitis (AE) Outbreaks in Brazil

“…Avian encephalomyelitis is a disease distributed worldwide (Tannock & Shafren, 1994;Boros et al, 2014;Haryanto et al, 2016;Welchman et al, 2009) and it has been described in Brazil in the state of Rio de Janeiro in 1692 (Chagas et al, 1962) and in the state of São Paulo in 1964 (Bueno et al, 1964). In 2015, an increase in cases of Avian Encephalomyelitis was reported; they were diagnosed through histopathology and serological analysis in the southern region of Brazil between 2012 and 2013 (Freitas & Back, 2015).…”

Detection of avian encephalomyelitis virus in chickens in Japan using RT-PCR

“…Clinically, AEV infection reveals similar symptoms as several other diseases, such as Newcastle disease, Marek’s disease, rickettsial diseases, vitamin B1 or B2 deficiency, aspergillosis, salmonellosis, coccidiosis, omphalitis, and mycoplasmosis [1, 9]. For the differential diagnosis, isolation and identification of the causal agent are extremely important.…”