1 1735-5680 Iranian Society for Virology 331 Special Evaluation of Epstein - Barr virus Frequency in Paraffin-Embedded Tissues of Hodgkin and Non-Hodgkin Lymphoma Patients in Kerman Province Pourrezaei S b jafarpour A c Aghamiri Sh d Molai Hamid Reza e Arabzadeh Ali Mohammad f Shahani T g b Health Research Institute, Infectious and Tropical Diseases Research Center, Kerman University of Medical Sciences, Kerman, Iran c Virology Division, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran d Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran e Virology Department, School of Medicine, Kerman University of Medical Sciences, Kerman, IR Iran f Virology Department, School of Medicine, Kerman University of Medical Sciences, Kerman, IR Iran g Virology Department, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, IR Iran 1 12 2017 11 4 1 7 14 08 2018 26 01 2019 Background and Aims: EBV infection usually occurs in humans and is very common. Sero-epidemiological surveys show that over 95% of adults worldwide are faced with EBV virus. The Epstein-Barr virus causes infectious mononucleosis and its association with a number of human cancers, including Hodgkin's lymphoma, non-Hodgkin lymphoma (especially lymphoma BL), nasopharyngeal carcinoma, gastric carcinoma and breast cancer has been established. Materials and Methods: This study was conducted cross-sectional. This study was designed to determine the presence of the Epstein-Barr virus genome in tissue samples of patients with Hodgkin lymphoma and non-Hodgkins. In this study, 40 samples of patients with non-Hodgkin's lymphoma and Hodgkin's disease, while had been kept in archives of the Shahid Bahonar Hospital, Afzalipour Hospital and Payambar-e-Azam Hospital of Kerman were examined by real-time PCR technique. The data collected by SPSS software and chi-square test were analyzed. Results: The prevalence of EBV in patients in this study was 5/27% of the 40 sample, of which 20 cases were of Hodgkin lymphoma and 20 cases were of non-Hodgkin lymphoma. No significant correlation was found between the prevalence of EBV in Hodgkin and non-Hodgkin lymphoma. There was also no correlation between men and women of different ages for the presence of EBV DNA in patients samples (P>0.05). Conclusions: EBV DNA associated with the tumor could be detected by molecular methods which are useful for the diagnosis of EBV-associated diseases, also biological factors such as age, sex, health status, social and economic factors in the pathogenesis of EBV and its relationship to lymphoma was not observed.
334 Special Applying of Recombinant Major Coat Protein for Production of Specific Antibody and Efficient Detection of Citrus Tristeza Virus (CTV) Samiei F h Safarnejad MR i Hosseini Salekdeh Gh j Shams- Bakhsh M k Safarpour H l h Department of Plant Pathology, Science and Research Branch, Islamic Azad University, Tehran, Iran i Department of Plant Viruses, Iranian Research Institute of Plant Protection, Agricultural Research Education and Extension Organization of Iran (AREEO), Tehran, Iran j Department of Genomics, Agricultural Biotechnology Research Institute of Iran, Karaj, Iran k Department of Plant Pathology, Faculty of agriculture, Tarbiat Modares University, Tehran, Iran. l Cellular and Molecular Sciences Research Center, Birjand University of Medical Sciences, Birjand, Iran 1 12 2017 11 4 8 15 03 11 2018 26 01 2019 Background and Aims: Citrus tristeza virus (CTV) is the most economically important pathogen of citrus throughout the world. To avoid the ominous effect of disease in citrus growing area, producing of virus free plants and elimination of infected plants are imperative. For this aim obtaining simple and sensitive diagnosis tools is crucial. The main objective of present study is applying of recombinant protein technology for production of specific antibody and developing of serological assays for efficient detection of CTV within infected plants. Materials and Methods: The major coat protein (p25) of CTV was selected as a target for preparation of polyclonal antibody. The gene encoding p25 was recombinantly expressed in bacterial host and the protein was purified through affinity chromatography approach. The purified recombinant coat protein was used for immunization of rabbit. Specificity of the prepared serum against CP was confirmed through serological assay. The immunoglobulin molecules were purified from serum through staphylococcus protein A followed by conjugation to alkaline phosphatase (AP) and horse radish peroxidase (HRP) enzymes. The prepared antibodies and conjugates were used for detection of infected plants by double antibody sandwich- enzyme linked immunosorbent assay (DAS-ELISA) and dot-blot immune binding assay (DIBA). Results: The p25 protein was expressed in bacterial host. The SDS-PAGE results confirmed high purity and integrity of CTV major coat protein with the expected size of about 29 kDa. The indirect ELISA results revealed that the antibody titer was around 1:65000. The IgG molecules purified through protein A column and SDS-PAGE results confirmed purity of the prepared antibody. The concentration of IgG was quantified by comparison to standard protein, BSA, which was estimated around 1 mg/ml.  The results obtained from DAS-ELISA and DIBA assays proved that prepared antibodies could be effectively applied for detection of CTV infected plants.   Conclusions: The prepared antibody and conjugates were powerful tools for detection of infected plants. To the best of our knowledge this is the first work for applying of peroxidase enzyme in developing of ELISA assay against CTV. 337 Special Sequence Analysis of the Spike Glycoprotein, Nucleocapsid Gene and 3' Untranslated Region of Razi Institute H-120 & H-52 Vaccine Strains of Infectious Bronchitis Virus khalesi B m Masoudei Sh n Bakhshesh M o Shoshtari AH p m Department of Research and Production of Poultry Viral Vaccine, Razi Vaccine and Serum Research institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran. n Department of Research and Production of Poultry Viral Vaccine, Razi Vaccine and Serum Research institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran. o Department of Research and Production of Poultry Viral Vaccine, Razi Vaccine and Serum Research institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran. p Department of Research and Production of Poultry Viral Vaccine, Razi Vaccine and Serum Research institute, Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran. 1 12 2017 11 4 16 24 30 11 2018 26 01 2019 Background and Aims: Infectious bronchitis (IB) is an economically important disease of chickens. The existence of very large number of IBV serotypes and variants which insufficiently induce cross protection against each other is the major problem to control the disease. Materials and Methods: This study was performed to characterize S1, N and 3'UTR region of the genome of H-120 and H-52 vaccine strains of Razi Vaccine Institute. The S1, N and 3'UTR region were sequenced and compared with standard strain in gene bank. Results: Based on nucleotide identity, the S1, N and 3'UTR region of the genome of Iranian IBV vaccine strain showed 100% similarity to the commonly standard IBV strains. To better characterization of these strains analysis of other genes involved in virulence and pathogenesis of the virus and performing protective tests against field strains are recommended. Conclusions: The results revealed that H-52 and H-120 strains of Razi institute were identical to the standard of strains in the Gene Bank. 344 Special Preparation of a pcDNA3 Plasmid for Hemagglutinin-Neuraminidase Gene of Human Respirovirus 3 (HPIV-3) Behzadpour D Alizadeh R Bamdad T Daliri M Seidavi A Department of Virology, Rasht Branch, Islamic Azad University, Rasht, IR Iran. Department of Genetics, school of medicine, Iran University of medical science, Tehran, IR Iran Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, IR Iran National Institute for Genetic Engineering and Biotechnology, Tehran, IR Iran. Islamic Azad University, Rasht Branch, Rasht, Iran 1 12 2017 11 4 25 30 15 12 2018 26 01 2019 Background and Aims: Human respirovirus 3 is the second most common agent for respiratory disease in children under five years old. Near 68% of respiratory tract infections belong to this type. However, there is no established vaccine for this virus. The aim of this study was to synthesize a DNA plasmid for HN gene of human respirovirus 3 (HPIV-3) as a first step for the preparation of a vaccine. Materials and Methods: The HN gene was synthesized and cloned into pcDNA3 plasmid. The plasmids were transformed into COS-7 cell lines. To evaluate the expressed HN protein, the western blotting assay was applied. Results: The HN protein 36 KD was detected on nitrocellulose membrane using anti-His-tag antibodies. Conclusions: We prepared a DNA plasmid containing HN gene of human respirovirus 3 to be used for the future studies towards making an effective vaccine. Among hospitalized Iranian children, one-month to five-year, human respirovirus 3 was most frequently observed in respiratory tract infection. 335 Special Production and Evaluation of Polyclonal Antibody Against Influenza A Virus Matrix 2 Conserved Protein for Research and Diagnosis Purposes Zamani S Fotouhi Chahouki F Mehrbod P Sadeghi Neshat S Farahmand B Department of Influenza and other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran/Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran Department of Influenza and other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran Department of Influenza and other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran Department of Influenza and other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran Department of Influenza and other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran 1 12 2017 11 4 31 38 16 11 2018 26 01 2019 Background and Aims: The aim of this study was to produce and purify the Polyclonal antibody (pAb) against Matrix protein 2 (M2) with reasonable efficiency. Matrix protein 2 is one of the most conserved proteins of the influenza A virus which acts as ion channel. Polyclonal antibodyproduced against Matrix protein 2 is used in vaccine research, passive immunization and qualitative/quantitative analysis methods. Materials and Methods: Recombinant M2 protein was produced in E.coli. Purified protein with Freund’s adjuvants (Complete and Incomplete) was injected into two New-Zealand white male rabbits. The polyclonal antisera of rabbits were evaluated by RID, immunoblotting and ELISA. The IgG was purified using DAEA-cellulose column chromatography. Finally, the quality and properties of purified IgG were evaluated using SDS-PAGE and ELISA. Results: The RID and immunoblotting results showed that the produced anti-M2 antibody was able to recognize M2 recombinant protein epitopes. The ELISA results confirmed anti-M2 pAb reached reasonable titers after three injections. IgG against M2 was purified with suitable concentration. The Purified polyclonal IgG-M2 was evaluated using ELISA and the results showed IgG-M2 reacted with the antigen up to 1:32000. Conclusions: The data showed that recombinant M2 protein was able to stimulate immune response to produce antibody at satisfactory level. 328 Special Identification of Respiratory Syncytial Virus (RSV) Genome in the Stool of a Child with Acute Gastroenteritis Akbari A Mohammadi J sadeghi k Azizi Jalilian F Abadan School of Medical Sciences, Abadan. Iran Department of Infectious Disease, Faculty of Medicine, Ilam University Of Medical Sciences, Ilam, Iran. Virology Department, School of Public HealthTehran University of Medical SciencesTehran Iran Department of Medical Microbiology, Faculty of Medicine, Ilam University of Medical Sciences,Ilam, Iran. Department of Virology, Faculty of Medicine, Hamadan University of Medical Sciences,Hamadan, Iran 1 12 2017 11 4 39 41 24 06 2018 26 01 2019 Background and Aims: Some viruses have been reported to cause respiratory and gastroenteric infections simultaneously. In this case we presented detection of human respiratory virus (RSV) type B genome from diarrheal sample of a 12 months' child with acute gastroenteritis. Results: The results indicated the presence of RSV subtype B genome in all three stool samples. Moreover, no sign of co-infections with other enteropathogenic agents reported. Conclusions: It could be a simple shedding of virus through gastroenteric system.