1 1735-5680 Iranian Society for Virology 315 General Perspective on Possible Recombination Event in Fusion Protein Gene of Newcastle Disease Viruses Isolated in Iran Shahsavandi Shahla b Ebrahimi Mohammad Majid c Ghodsian N b Razi Vaccine & Serum Research Institute c Razi Vaccine & Serum Research Institute 1 6 2017 11 2 1 7 02 12 2017 15 04 2018 Background and Aims: Newcastle disease (ND), caused by the virulent Newcastle disease virus (NDV), is one of the most important viral diseases in birds. In recent years recombination occurring throughout the NDVs genome isolated in China and Indonesia has been reported. This study was focused to investigate the recombination events in the F gene of the Iranian NDVs to generate useful data that can be applied in controlling programs. Materials and Methods: Sequences of 1200 base pairs of the F gene of Iranian NDVs were analyzed compared with the isolates from other reference strains. Divergence between the gene sequences and the influence of natural selection were estimated. The neutralizing epitopes and potential N-glycosylation sites within F proteins were determined. Possible recombination within the sequences was analyzed using RDP3 software. Results: Alignment and phylogenetic analysis based on the F gene revealed that Iranian NDVs share a higher nucleotide identity with NDVs representing genotype VII and further clustered into two sub-genotypes. The calculated Ka/Ks and negative Tajima’s D test indicate purifying/stabilizing selection. No recombination events were detected in F gene of Iranian NDV sequences deposited in GenBank. Conclusions: While no recombination event was identified for the gene, constant molecular and pathological characterization of circulating NDVs are needed to detect an evolutionary feature of the viruses.
316 General Low Prevalence of Borna Disease Virus RNA in Patients with Bipolar Major Depression and Schizophrenia in North of Iran Mohammadi Manesh Maryam Mohebbi Alireza Yasaghi Mohammad Najafi Memar Zahra Javid Naeme Taziki Sadegh Ali Kalani Mohamad Reza Tabarraei Alijan Moradi Abdolvahab 1 6 2017 11 2 8 13 02 12 2017 15 04 2018 Background and Aims: Borna disease virus (BDV) is well known as a neurotropic virus, however, its role in human neurological diseases such as schizophrenia and bipolar disorder is still unclear. In this study, we aimed to investigate the BDV genome in such patients in Golestan province, North of Iran. Materials and Methods: RNA was extracted from peripheral blood mononuclear cells (PBMCs) from 250 people (125 patients with bipolar disorder (94.5%) and schizophrenia (3.1%) and 125 sex and age-matched healthy blood donors). Demographic information was collected through a questionnaire. RNA-driven cDNA was used for further BDV P40 genome tracking by Polymerase Chain Reaction (PCR). Results: Only one sample (1/125; 0.8%) of a female patient with bipolar disorder found to be positive for BDV P40. No significant family history of the disease found in both groups of patients. Seven patients with bipolar depression disorder (5.5%) had a history of animal contact. Conclusions: No BDV genome has been detected in the blood samples of the patients admitted to the hospital section of psychiatry in Gorgan city. Despite other genetic and environmental factors involved in psychiatric disorders, the serological study will give us a better insight of BDV prevalence in the region. 317 General Identification of RD and L20B Cell Lines Cross-Contamination by Multiplex SYBR-Green PCR in Poliovirus Laboratory Nejati A n Tabatabaeie H o Yousefi M p Mollaei-Kandelous Y Shahmahmoodi Sh n Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595 o Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595 p Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595; Department of Virology, International Campus, Tehran University of Medical Sciences, Tehran, Iran. Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595; Immunology Department, Iran University of Medical Science, Tehran 14716-13151, Iran. Polio National Laboratory, Department of Virology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, telephone number: +982188950595; Food Microbiology Research Center, Tehran University of Medical Sciences, Tehran, Iran. 1 6 2017 11 2 14 18 03 12 2017 15 04 2018 Background and Aims: World Health Organization Global Polio Laboratory Network (GPLN) plays a critical role in the Global Polio Eradication Initiative. Cell culture methods (mostly RD and L20B cell lines) have been used for Enteroviruses and polioviruses isolation, respectively. Cross-contamination among L20B and RD cell lines causes the problem in accuracy of poliovirus surveillance and decreases the poliovirus detection. Therefore, validation of identity of cell lines purity is a vital part of cell culture in polio laboratory. Materials and Methods: In this study, a multiplex SYBR-Green PCR based on Cytochrome b oxidase amplification was designed to L20B and RD cell lines cross-contamination. Results: The conventional multiplex PCR performed on DNA extracted from L20B cells deliberately cross-contaminated with RD cells clearly showed not only the identity of L20B cell line but also the presence of contaminant RD cells. Conclusions: The results indicated that the multiplex SYBR-Green PCR was reliable method to identity L20B and RD cell lines individually and also after deliberate cross-contamination. 341 Special Sequence and Phylogenetic Analysis of Membrane (M) Gene of Infectious Bronchitis Viruses Isolated in Iran during 2014 - 2015 Kami E Ghalyanchilangeroudi A Madadgar O Nayyeri Fasaei B Hosseini H Najafi H Mousavi FS Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Poultry diseases Faculty of Veterinary Medicine, Azad University, Karaj, Iran Department of Pathobiology, School of Veterinary Medicine, Shiraz University Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran 1 6 2017 11 2 19 28 10 12 2018 10 12 2018 Background and Aims: Avian infectious bronchitis virus (IBV) has a worldwide distribution and mutations occurring in the large viral genome of IBV have led to extensive antigenic variations among IBVs. This is the first study conducted to determine the complete membrane (M) gene sequences of different Iranian IBV genotypes. Materials and Methods: The M gene of three 793/B (IBKG1,6,7), one Massachusetts (IBKG3), three Variant 2 (IBKG2,4,9), four QX(IBKG5,10,11,12) and one IR-1(IBKG8) Iranian IBV isolates were sequenced and analyzed. Also, post-translational modifications of the M glycoprotein were predicted. Results: Sequence analysis of the M gene indicated IBKG1, 6, 7, and 8 shared the highest homology to 793/B genotype. IBKG2, 4, 5, 9, 10, 11, and 12 showed the highest similarity to QX-like IBV. IBKG3 (Massachusetts) isolate shared 99/53% identities with H120 vaccine strain, respectively. Although M proteins are moderately well conserved within each coronavirus genus, the sequence differences of M gene reached to 12% among Iranian IBV isolates. Based on M gene analysis, IBKG-8 (IR-1 genotype) and all Variant2 IBVs, classified as a 793/B and QX IBV according to the M gene sequences, respectively. Conclusions: It is the first characterization of IR-1 (Iranian local genotype) based on M gene. Also, the data shows M gene of variant 2 IBV strains are derived from QX genotypes. The results complete the Iranian IBVs genetic puzzle. 342 Special The Antiviral Effects of Curcumin Nanomicelles on the Attachment and Entry of Hepatitis C Virus Naseri S Darroudi M Aryan E Gholoobi A Rahimi HR Ketabi K Movaqar A Abdoli M Gouklani H Teimourpour R Meshkat Z Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Nuclear Medicine Research Center (NMRC), Ghaem Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Department of Modern Sciences and Technologies, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Department of Modern Sciences and Technologies, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran Molecular Medicine Research Center, Hormozgan University of Medical Sciences, Bandar Abbas, Iran Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran 1 6 2017 11 2 29 35 10 12 2018 10 12 2018 Background and Aims: Hepatitis C virus (HCV) is a member of the Flaviviridae family, which causes approximately 500,000 deaths annually. HCV infection treatment is often associated with significant adverse effects. Curcumin is an active ingredient of turmeric which has therapeutic anti-inflammatory effects in many diseases including infectious ones. Although curcumin is not soluble in water, if it is synthesized in the form of nanomicelles, it will be water soluble and can be absorbed in the gastrointestinal tract (GI). In this study, the antiviral effects of curcumin nanomicelles were investigated on the attachment and entry of HCV particles. Materials and Methods: The cytotoxicity of curcumin nanomicelles was determined in Huh7.5 cells and their antiviral effects on the attachment and entry of HCV was investigated in a cell culture system. Results: Curcumin nanomicelles could decrease the viral load in the cell culture supernatants compared to virus control. Conclusions: According to the results of this research, we determined the antiviral effects of curcumin nanomicelles in the later stages of HCV replication. 322 General Polyclonal Antibody against Recombinant Nucleoprotein of the Influenza A Virus (H1N1); Production and Purification Alikhani Mehdi Behzadian Farida Mehrbod Parvaneh Khosravi Node Fatemeh Shokouhi Targhi Hadiseh Farahmand Behrokh 1 6 2017 11 2 36 42 13 02 2018 15 04 2018 Background and Aims: Influenza is an acute respiratory illness that is caused by a virus belonging to Orthomyxoviridae family. This virus spreads rapidly every year in cold season and leads to morbidities and mortalities especially in adults and children, which causes billions of dollars of economic losses. Accordingly, development of a rapid, sensitive and inexpensive laboratory diagnosis based on antigen detection to distinguish this infection from other respiratory tract viruses is important. In addition, specific anti-influenza antibody production against influenza virus antigen is essential for basic and applied research programs. Influenza A virus nucleoprotein (NP) is a structural protein and a major component of the ribonucleoprotein complex. It has a high expression level during infection. NP consists of 498 amino acids with molecular weight of 57 KD. The aim of this study was to produce and purify polyclonal antibody against recombinant nucleoprotein of the influenza A virus. Materials and Methods: Rabbit immunization was performed based on a specific program by NP purified recombinant antigen and Freund's adjuvant. Serum immunoglobulin was separated by ammonium sulfate and IgG purification was conducted by ion exchange chromatography (DEAE-cellulose). To evaluate the reaction between antigens and purified antibodies, SRID and ELISA serological tests were applied. Results: The results obtained from SDS-PAGE and Western blot showed a dense band of purified NP. The results of ELISA confirmed an increase in NP antibody titer after one month. Antibody levels detection by ELISA showed a sensitivity of 1 to 50. In SRID, sedimentary areola was observed due to the interaction of NP antigen and antiserum. Western blot results were also positive for the NP protein. Conclusions: The NP antigen purified in this study, as well as the produced and purified antibodies, had the ability to be used in serological tests to detect influenza A virus. It can also be used in basic research methods such as Western blot, immunohistochemistry and immunocytochemistry. Keywords: antigen purification, polyclonal antibody production, antibody purification,