1 1735-5680 Iranian Society for Virology 83 General Seroprevalence of Cytomegalovirus Infection in Blood Donors in Khorramabad Delfan-Beiranvand M b Pournia Y c Fazeli M d Tarvand AH e Hosseini J f Mirzaei M g b Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Khorramabad, Iran c Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran d Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran e Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Khorramabad, Iran f Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Khorramabad, Iran g Department of Laboratory Sciences, Borujerd Branch, Islamic Azad University, Borujerd, Iran 1 2 2012 6 1 1 5 07 11 2014 07 11 2014 Background and Aims: Cytomegalovirus (CMV) has worldwide distribution, and its prevalence rate depends on factors such as economic and geographical conditions. An important way of the virus transmission is via blood. Due to high prevalence of anti-CMV antibodies in blood donors and lack of data concerning its seroprevalence in the region, this study was carried out to determine the prevalence rate of anti-CMV antibodies in the blood donors of Khorramabad Blood Transfusion Center. Materials and Methods: This descriptive study was conducted on 270 healthy donors referring to Khorramabad Blood Transfusion Center. The demographic data were recorded in the questionnaires, and following the routine screening tests, the tests of anti-CMV antibodies (IgG and IgM) were performed using the Iranian Pishtaz-teb kit through the ELISA technique. The data were analyzed by the t-test and x2 test using the SPSS software. Results: Out of 270 samples, 90% were males, and 10% were females. Anti-CMV IgG antibody was positive in 148 samples (55%), and negative in 122 ones (45%). Moreover, anti-CMV IgM antibody was negative in 269 cases (99.6%), and positive in 1 case (0.4%). Conclusion: Considering the high seroprevalence of anti-CMV antibody in Khorramabad, latency of the virus inside the blood cells, and its possible transmission via blood and blood products to blood receivers particularly in immunodeficient patients including those with malignant diseases receiving chemotherapy and recipients of allograft transplants, performing screening tests on donated blood samples for CMV infection particularly in high risk cases is recommended.
84 General Determination of Hepatitis Delta Virus Genotype among HBV Carriers in Southwest of Iran Khalafkhany D h Makvandi M i Javanmard D j Hamidi-Fard M k Samarbaf-Zadeh AR l h Department of Virology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran i Department of Virology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran j Department of Virology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran k Department of Virology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran l Department of Virology, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 1 2 2012 6 1 6 12 07 11 2014 07 11 2014 Background and Aims: HDV is a defective satellite virus and classified in genus Deltavirus. Its disease is related and limited to HBV-infected patients. Acute infection of delta agent occurs in two different patterns simultaneous infection with both HBV & HDV or super infection of chronically HBV infected patients that lead to more sever type of hepatitis. According to genetic diversity of genomic RNA of HDV, 8 clades have been classified. The aim of this study was to determine the delta virus genotype among the HBV & HDV infected patients in Ahvaz city. Materials and Methods: Sera sample collected from 31 seropositive HDV patients including 21 male and 9 female mean age 46±13.5 suffering from chronic hepatitis and 1 male patient with acute hepatitis. The encoding region for C terminal half of the Delta anti gene of the HDV’s genomic RNA reverse transcribed and then amplified by nested PCR. The HDV PCR positive samples were sequenced, and the sequences were compared with reference sequences on GenBank. All samples were tested for HBV DNA, HCV RNA and HCV anti body. Results: Only 15(48%) out of 31 anti HDV seropositive patients were positive for HDV RNA by nested RT-PCR. Alignment and phylogenic analysis of the present HDV sequences revealed that all sequences belong to clade 1. Only 1 HDV RNA positive patient was positive for HBV DNA by nested PCR. Conclusion: According to previous studies the clade 1 (genotype 1) is the predominant clade of HDV in our country. However some (2-HDV-R, 4-HDV-R &11-HDV-R) of our isolates show extensive differences from the two previously isolated HDVs from Iran. Most of the our isolates were closely related to the Iranian HDVs, but Egyptian HDV still remains the most relevant foreign isolate. Suppression of HBV replication by delta virus is common but mutual suppression of HDV and HCV remains unclear. 85 General Expression of Influenza Heamagglutinin Globular Head in Different Eukaryotic Cells Tavakoli R m Fotouhi F n Tavassoti-Kheiri M o Farahmand B p Saleh M Tabatabaeiyan M m Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran n Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran o Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran p Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran 1 2 2012 6 1 13 17 07 11 2014 07 11 2014 Background and Aims: Influenza (flu) is a respiratory infection in mammals and birds. It is caused by an RNA virus in the family Orthomyxoviridae. The virus is divided into three main types. Influenza virus type A is found in a wide variety of bird and mammal species and can undergo major shifts in immunological properties. Hemagglutinin (HA) is an important influenza virus surface antigen that is highly topical in influenza research. In the present study, the gene encoding HA1 protein which includes Hemagglutinin globular head from influenza virus A/Tehran/18/2010 (H1N1) was cloned into a eukaryotic expression plasmid (pCDNA3) and its expression was evaluated in eukaryotic cells. Materials and Methods: HA1 gene was incised from pFastBacTHc-HA1 by digestion, purified and subcloned into eukaryotic expression vector (pCDNA3). After verification of the cloning fidelity, the recombinant plasmid was transfected into COS-7 and BHK-21 cells, and its expression was detected by RT-PCR. Results: Restriction endonuclease digestion analysis, colony PCR and DNA sequencing indicated that the recombinant plasmid pCDNA3-HA1 had been constructed successfully. After transfection into eukaryotic cells, the presence of mRNA transcripts was verified by reverse transcriptase-polymerase chain reaction (RT-PCR). Conclusion: This study is a demonstrated success in the construction of eukaryotic expression plasmid for HA1 thus providing a basis for further probing into mechanism of virus infection and exploring DNA vaccine. 86 General Phylogenetic Analysis of Hemagglutinin Gene of H9N2 Avian Influenza Viruses Isolated from Chicken in Iran in 2010-2011: Emerging of a New Subgroup Norouzian H Gholami SJ Vasfi-Marandi M Department of Clinical Sciences, Faculty of Veterinary Medicine, Lorestan University, Khorram Abad, Iran Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran 1 2 2012 6 1 18 26 07 11 2014 07 11 2014 Background and Aims: Hemagglutinin (HA) protein of Avian Influenza (AI) plays an essential role in the virus pathogenicity. AI H9N2 subtype causes significant economic loss in broiler and layer in poultry farms in Iran. AI viruses have a great involvement in evolutionary changes at nucleotide and amino acid levels and vaccines could induce faster rates of such changes. Up-dated understanding of the genetic changes of AI viruses circulating in Iran is necessary for controlling AI. Materials and Methods: Sequence analysis and phylogenetic study of the HA gene of three H9N2 subtype of AI isolates in Iran in 2010-2011 were studied. Results: Cleavage site of the Iranian 2010-2011 isolates possessed a different motif. Amino acid residue at position 226 at receptor binding site in these isolates was Leucine, which was similar to human viruses. The epitopes for HA showed a great variation related to the year of isolation. According to phylogenetic analysis, Iranian isolates were divided into two main subgroups. But, viruses isolated in this study formed a third minor subgroup. Degree of homology between the 2010-2011 isolates and former Iranian isolates was significantly low. Conclusion: The results revealed that HA of new Iranian AI H9N2 isolates have undergone extensive genetic changes. Definitely, continuous monitoring of genetic changes is a useful tool for updating control strategy for AI outbreak in Iran. 87 General The Effect of High Temperature Treatment on Wheat Streak Mosaic Virus Resistance and Certain Resistance-Related Chemicals in Bread Wheat Zinati Z Assad MT Masumi M Alemzadeh A Razi H Izadpanah K Department of Plant Production and Plant Breeding, College of Agriculture, Shiraz University, Shiraz, Iran Department of Plant Production and Plant Breeding, College of Agriculture, Shiraz University, Shiraz, Iran Agricultural Research Center of Fars and Plant Virology Research Center, Shiraz University, Shiraz, Iran Department of Plant Production and Plant Breeding, College of Agriculture, Shiraz University, Shiraz, Iran Department of Plant Production and Plant Breeding, College of Agriculture, Shiraz University, Shiraz, Iran Plant Virology Research Center, College of Agriculture, Shiraz University, Shiraz, Iran 1 2 2012 6 1 27 35 07 11 2014 07 11 2014 Background and Aims: To evaluate the effect of temperature on wheat streak mosaic virus (WSMV) resistance phenotype, through total protein, phenol, and peroxidase activity in bread wheat, a factorial experiment was conducted using Adl-Cross (resistant) and Marvdasht (susceptible) cultivars. Materials and Methods: Results showed that incubation at 32C changed the gene expression for resistance to WSMV and mosaic symptoms were observed in Adl-Cross. Total protein reduction in inoculated Adl-Cross was significant at 32C. Results also indicated that high temperature either prevented expression of genes or degenerated available proteins involved in resistance mechanism. Total protein in infected Marvdasht was significantly reduced as compared with healthy control plants. Since electrophoretic pattern indicated reduction of ribulose 1, 5-bisphosphate carboxylase (RBPC) subunits in infected Marvdasht, reduction of protein may have probably been due to a decrease in the synthesis of RBPC. Mean of phenolic compounds content in Adl-Cross was higher as compared to Marvdasht in both infected and non-infected plants. Total phenol increased 2.8 and 4.06 percent in inoculated Marvdasht and Adl-Cross, respectively. The trend of increase in phenolic compounds indicated that their synthesis and accumulation was higher in Adl-Cross as compared to Marvdasht. Results: These results indicated the role of phenolic compounds in prevention of viral movement and spread in resistant cultivar. Thin-layer chromatography (TLC) analysis showed that the intensity of a spot with Relative flow (Rf) 0.622 increased at high temperature. Increase in total phenol at high temperature may have been due to increase in intensity of this spot. Spot concentration with Rf 0.622 at high temperature was higher in infected samples as compared to uninfected samples. Conclusion: This showed an interaction of virus and temperature. Also, a spot with the same Rf and different color was observed 8 days after inoculation at 25C. The color change in this spot showed that high temperature might cause a decrease in concentration of phenolic compounds which are in turn effective in resistance to WSMV. Another possibility is that the compounds effective in resistance of Adl-Cross are changed to neutral forms at higher temperatures. There were no significant differences between genotypes for peroxidase activity in healthy plants at 25°C. Viral infection reduced the peroxidase activity in Marvdasht but, showed a significant increase in Adl-Cross. High temperature reduced peroxidase activity in both infected and uninfected plants. Peroxidase enzyme probably affects synthesis of compounds effective in resistance. Also, reduction in enzyme activity at high temperature increased reactive oxygen species (ROS) and this led to oxidative stress. 88 General Nucleoprotein (NP) Gene Based Phylogenetic Analysis of Iranian H9N2 Avian Influenza Isolates during 1998-2011 Majidzadeh- A K Soleimani M Karimi V Shojaee-Estabragh A Fanni A Mandegar O Ghalyanchi-Langeroudi A Iranian Centers for Breast Cancer (ICBC); Academic Center for Education, Culture and Research (ACECR), Tehran, Iran TASNIM Biotechnology Research Center, AJA University of Medical Sciences, Tehran, Iran Departments of Clinical Sciences, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran QC Expert ,Pasteur Institute of Iran, Tehran, Iran Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran 1 2 2012 6 1 36 42 07 11 2014 07 11 2014 Background and Aims: Avian Influenza (AI) H9N2 subtype was first reported to infect turkeys in the United States in 1966 and has been panzootic in Eurasia. In Iran, the H9N2 virus was first isolated from broiler chickens in 1998 in Ghazvin province and it is the most prevalent subtype of influenza virus in poultry industry in Iran at the present time. Materials and Methods: In this study, we sequenced and analyzed Nucleoprotein (NP) gene of six AI H9N2 isolates from broiler farms of different parts of Iran from 1998 to 2011 to show probable changes since first advent. Results: Results indicate that nucleotide homology among these isolates with NP genes is between 91.8% to 98.8%. The divergences between isolates have significantly been increased since 2007.Iranian AI H9N2 Isolates based on NP gene divided in two distinct clusters according to their isolation year. Group 1 is located in Y-439 clade and Group 2 is located in G1 Clade. Iranian H9N2 isolates of avian influenza virus show more amino acid substitutions Compare to those found in human H9N2 isolates. Conclusion: The results shown here that further gene reassortment has occurred subsequent to the emergence of viruses in the Middle East highlights the potential for viruses to evolve rapidly.