ORIGINAL_ARTICLE Background and Aims: Despite the screening of blood donors, blood transfusion represents an ideal port of entry for blood-borne infection. Blood-borne pathogen transmission has been a concern since the earliest days of transfusion. The blood product of platelet (PLT) concentrates is still faced with the risk of bacterial and viral contaminations. Pathogen inactivation technologies offer a proactive approach and the potential to further improve blood safety. Here we study the pathogen inactivating capacity of riboflavin with UV light treatment in platelet concentrates contaminated with enveloped and non-enveloped viruses. Materials and Methods: The inactivation effects of riboflavin in combination with UV light was examined on Herpes simplex virus (HSV), Vesicular stomatitis virus (VSV) and Polio virus classified as enveloped and non-enveloped DNA and RNA viruses, respectively. After spiking viruses in PLT concentrate, treatment was undertaken with riboflavin (50 µM) and exposed to different doses of UV light. Residual viral infectivity was titrated using 50%Tissue Culture Infective Dose (TCID50). Results: Combination of Riboflavin and UV light treatment reduced the titer of HSV, VSV and Poliovirus with different doses of UV light. Log reduction of HSV with UV doses of 0.82, 1.63, 2.44 and 3.25 J/cm2 was 0.5, 1.3, 3.5 and 3.8, respectively. Log reduction of VSV for 0.82, 1.63, 2.44 and 3.25 J/cm2was 2.8, 3.8, 4.6 and 5.6, respectively. Also, log reduction of poliovirus in the same order for 0.82, 1.63, 2.44 and 3.25 J/cm2was 1.7, 2.5, 2.7 and 3.1 respectively. Conclusion: The final dose of UV (3.25 J/cm2) resulted in a larger amount of viral inactivation for enveloped and non-enveloped viruses, in suspended PLT. This method offers a potential to be used for prevention of the majority of PLT transfusion-associated viral pathogens. http://journal.isv.org.ir/article-1-111-en.pdf 2014-12-09 1 6 10.21859/isv.7.1.2.1 Riboflavin UV light Platelet concentrate Virus inactivation Hamideh Mirshafiee 1 Department of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, Iran AUTHOR Seyed Masoud Hosseini 2 Department of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, Iran AUTHOR Zohreh Sharifi 3 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran AUTHOR Hamid Latifi 4 Laser and Plasma Research Institute, Shahid Beheshti University, Tehran, Iran AUTHOR Fatemeh Yari 5 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran AUTHOR Hamed Nikbakht 6 Laser and Plasma Research Institute, Shahid Beheshti University, Tehran, Iran AUTHOR Amehen Elikaei 7 Department of Microbiology, Faculty of Biological Sciences, Shahid Beheshti University, Tehran, Iran AUTHOR

ORIGINAL_ARTICLE Background and Aims: The novel approaches in influenza vaccination have targeted more conserved viral proteins such as nucleoprotein (NP) to provide cross protection against all serotypes of influenza A viruses. Influenza specific cytotoxic T lymphocytes (CTL) are able to lyse influenza-infected cells by recognition of NP, the major target molecule in virus for CTL responses. On the other hand, studies suggest that fusing of molecular adjuvants such as Heat Shock Protein 70 (HSP70), member of intracellular chaperon super families, with an antigen can induce the cellular and humoral specific responses better than the same induced by antigen alone. It is shown that the C-Terminal of HSP70 (ctHSP70) is the main domain responsible for inducing immunity system. Materials and Methods: In this study the open reading frame of NP gene from Influenza A virus (PR/8/34) and C-Terminal (359-625) domain of HSP70 gene from Mycobacterium tuberculosis were amplified and cloned into expression pET28a vector independently. Then the N-terminal of whole NP protein was fused to truncated HSP70 in same vector. The fidelity of cloned genes was confirmed by sequencing. All three types of clones were expressed in E. coli BL21 and confirmed by SDS-PAGE and Western blot analysis. Results: Results showed the integrity of vector constructs and well expression of NP, ctHSP70 and fusion form of ctHSP70-NP recombinant proteins in BL21 host cells. Conclusion: ctHSP70-NP fusion protein produced could be considered and evaluated as a universal influenza vaccine which its immunogenicity potential needs to be assessed in animal models along with proper control groups including recombinant NP and ctHSP70 proteins. http://journal.isv.org.ir/article-1-112-en.pdf 2014-12-09 7 14 10.21859/isv.7.1.2.7 Influenza virus vaccine Nucleoprotein HSP70 Zahra Yousefi-Najafabadi 1 Department of Biosciences and Biotechnology, Malek-Ashtar Uniersity of Technology, Tehran Iran AUTHOR Farida Behzadian 2 Department of Biosciences and Biotechnology, Malek-Ashtar University of Technology, Tehran Iran AUTHOR Fatemeh Fotouhi-Chahooki 3 Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran AUTHOR Behrokh Farahmand 4 Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran AUTHOR Mahdi Tat 5 Applied Virology Research Center, Baqyatallah University of Medical Sciences, Tehran, Iran AUTHOR

ORIGINAL_ARTICLE Background and Aims: Newcastle disease virus (NDV) is one of the major pathogen in poultry. Vaccination is intended to control the disease as an effective solution nevertheless this virus is a growing threat to the poultry industry. F gene open reading frame (ORF) from NDV is 1650 bp, encoding a protein of 553 amino acids that can induce protective immunity alone. The F glycoprotein on the surface of NDV is important for virus infectivity and pathogenicity. Towards protection goal, the full-length of F gene was isolated using specific primers and cloned into the baculovirus derived bacmid shuttle vector to produce recombinant F-protein in insect cells. Materials and Methods: F gene ORF from avirulent strain La Sota of NDV was amplified by RT-PCR to produce F cDNA. The amplicon was cloned firstly into the T/A cloning vector and then subcloned into the pFastBac Dual donor plasmid through NcoI/KpnI sites. After the verification of cloning process by PCR and enzymatic digestion analysis, the accuracy of F gene ORF in the T/A cloning vector was confirmed by sequencing. Finally, F-containing recombinant bacmid was subsequently generated and confirmed by PCR using F specific primers and M13 universal primers. Results: Results showed that a recombinant baculovirus containing a correct and in framework sequence of Newcastle F gene under the control of p10 promoter was constructed. Conclusion: The above mentioned F-containing recombinant baculovirus, in addition to its independent application, can be used with other individual recombinant baculoviruses expressing NH and NP genes to produce Newcastle VLPs in insect cell line. http://journal.isv.org.ir/article-1-113-en.pdf 2014-12-09 15 20 10.21859/isv.7.1.2.15 Newcastle disease F protein Baculovirus Bacmid shuttle vector Mohammad reza Shafaati 1 Department of Microbiology, Islamic Azad University, Damaghan Branch, Damaghan, Iran AUTHOR Majid Moghbeli 2 Department of Microbiology, Islamic Azad University, Damaghan Branch, Damaghan, Iran AUTHOR Roholah Dorostkar 3 Applied Virology Research Center, Department of Virology, Baqiyatalah University of Medical Sciences, Tehran, Iran AUTHOR

ORIGINAL_ARTICLE Background and Aims: Accurate and rapid diagnosis is necessary for effective control and prevention of foot-and-mouth disease (FMD). In present study, was evaluated real time reverse transcription-polymerase chain reaction (rRT-PCR) assay along with diagnostic routine methods for the detection of all seven serotypes of FMD virus (FMDV), namely O, C, A, SAT1, 2, 3 and Asia 1 in biological samples at the reference laboratory for FMD, Iran. Materials and Methods: Two different RT-PCR assays targeting two different regions 5´ untranslated region (5´-UTR) and RNA polymerase (3D) of the FMDV genome were used to confirm the presence of FMDV in epithelial suspensions. Results: In the two methods the viral RNA in all tested archival serotypes of FMDV were detected. Specificity of this reaction was confirmed by the use of swine vesicular disease virus and blue-tongue. The amount of cycle threshold (CT) value of all seven serotypes was different and the lowest and highest of CT value achieved for SAT3, A, O types and SAT2, C types, respectively. Conclusion: The results showed that RT-PCR was more sensitive and effective than routine diagnostic methods. Furthermore, RT-PCR as a strong and valuable tool concomitant with diagnostic routine methods facilitate monitoring the fields FMDV strains and suggested that the use of the multiple diagnostic targets could enhance the sensitivity of the molecular methods for the detection of FMDV. http://journal.isv.org.ir/article-1-114-en.pdf 2014-12-09 21 29 10.21859/isv.7.1.2.21 Foot-and-Mouth Disease Virus ELISA real-time PCR conventional RT-PCR A Ahmadi-Vasmehjani 1 Department of Microbiology, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR SD Mousavi-Nasab 2 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran AUTHOR R Baharlou 3 Department of Microbiology, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR F Jeirani 4 Department of FMD, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR M Shayestehpour 5 Department of Virology, Tehran University of Medical Sciences, Tehran, Iran AUTHOR S Yaghoubi 6 Department of Microbiology, Tehran University of Medical Sciences, Tehran, Iran AUTHOR H Fazel 7 Department of Virology, Tehran University of Medical Sciences, Tehran, Iran AUTHOR H Mahravani 8 Department of FMD, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR

ORIGINAL_ARTICLE Background and Aims: The ectodomain of matrix protein of influenza virus is a weak immunogen that is highly conserved among all subtypes of influenza A virus. Tandem repeats of these genes along with linker were used to enhance immunogenicity of M2e protein and so it can be served as a universal vaccine in both humans and livestock. Materials and Methods: In this study, the sequences of extra-domain of matrix protein of influenza A registered in NCBI was converted into codons compatible for Bacillus subtilis using JAVA codon adaptation tool software. Results: A cassette consist of four repeats of this codon optimized sequence, spaced by appropriate linkers and flanked by BamHI and HindIII restriction sites was designed and thoroughly used for the synthesis. The cassette then was cloned into pMR12 shuttle expression vector. Conclusion: Two kinds of prokaryotic host, E. coli BL21and Bacillus subtilis WB600 were transformed by pMR12+4M2e. The fidelity of the construct in both transformants was confirmed by enzymatic analysis and PCR. http://journal.isv.org.ir/article-1-115-en.pdf 2014-12-09 30 36 10.21859/isv.7.1.2.30 Influenza A Matrix protein 2 Cloning Bacillus subtilis pMR12 Hamid Hoseinian 1 Department of Biology, Islamic Azad University, Damghan Branch, Semnan, Iran AUTHOR Meysam Moghbeli 2 Department of Biology, Islamic Azad University, Damghan Branch, Semnan, Iran AUTHOR Farida Behzadian 3 Department of Molecular Genetics, Research Center for Sciences and Biotechnology, Malek Ashtar University, Tehran, Iran AUTHOR

ORIGINAL_ARTICLE Background and Aims: Canine distemper (CD) is a deadly infectious disease of Canidae family. CD is a multi-systemic viral disease and is specified by wide range of clinical symptoms. The manifestations are not always indicative of CD, therefore a laboratory confirmation is necessary for suspected cases. Materials and Methods: Different clinical specimens of 19 CD suspected unvaccinated dogs were examined for canine distemper virus (CDV) infection by reverse transcription polymerase chain reaction (RT-PCR), Nested-PCR, and serum neutralization (SN) test during 2008-2011. RT-nested PCR assay was adjusted for detection of CDV nucleoprotein (NP) in prepared samples. Results: In samples of 3 out of 19 (15%) dogs, CDV NP gene was confirmed by RT-PCR while RT-PCR and combination with Nested-PCR (RT-nested PCR) presence of CDV NP gene was detected in various samples of 14 (73%) dogs. So efficiency of RT-PCR along with Nested-PCR raised 58%. Among different kinds of obtained samples, conjunctival swabs and kidney tissue biopsies were found to be suitable for analysis of CDV RNA. Additionally CDV antibody was detected in 11 out of 18 serum samples (61%) by SN test, but detection of neutralizing antibodies didn't comply with RT-nested PCR results. Conclusion: Results of this study indicated that Nested-PCR is a sensitive and applicable method for the diagnosis of CDV. http://journal.isv.org.ir/article-1-116-en.pdf 2014-12-09 37 43 10.21859/isv.7.1.2.37 Canine Distemper Virus RT-nested PCR Nucleoprotein Serum Neutralization Test S Namroodi 1 Department of Environmental Sciences, Faculty of Fisheries and Environmental Sciences, Gorgan University of Agricultural Sciences & Natural Resources, Gorgan, Iran AUTHOR A Rostami 2 Section of Zoo and Wildlife Medicine, Department of Internal Medicine, Faculty of Veterinary Medicine, University of Tehran AUTHOR K Majidzadeh-Ardebili 3 Faculty of Medicine, Army University of Medical Sciences, Tehran, Iran AUTHOR A Ghalyanchi-Langroudi 4 Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR A Morovvati 5 Department of Microbiology, Islamic Azad University of Qom- Branch, Qom, Iran AUTHOR

ORIGINAL_ARTICLE Background and Aims: Torque teno virus (TTV) is a DNA virus that isolated from the serum of a Japanese patient without A-G transfusion-transmitted hepatitis and other etiology. TTV is detectable in plasma and peripheral blood mononuclear cells, different body fluids and secretions such as stools, saliva, semen, vaginal fluid. The genome exhibits high diversity that has enabled the determination of several genotypes and subtypes and at least 30 genotypes have been identified. TTV might be seen as a risk factor in acute and chronic hepatitis, but it is not clear. Coinfection of TTV and HBV or TTV and HCV is common, because these viruses share the same transmission routes such as blood transfusion. The aim of this study was to determine the prevalence of TTV in patients with chronic HBV and HCV in the Southern of Iran and evaluate effect of TTV infection on the liver diseases. Materials and Methods: Serum samples collected from all hepatitis patients and healthy control subjects were included for serological tests for hepatitis B and C viruses. Briefly, DNA was isolated from serum of collected peripheral blood mononuclear cells (PBMC) of patients and carry out Semi Nested PCR Detection for TTV DNA. Results: These results showed the significant relationship between TTV and the patients that had chronic HBV and HCV (p<0.01). Conclusion: According to the result of this study, the prevalence of TTV in patients with chronic HBV and HCV in the southern of Iran was 50.8 and 66.5 %, respectively. These results were comparable to those reported in previous studies ranging from 20% to 75.7% for hepatitis C and from 40% to 75% for hepatitis B patients. http://journal.isv.org.ir/article-1-118-en.pdf 2015-01-06 44 50 10.21859/isv.7.1.2.44 Torque Teno Virus Coinfection HBV HCV SD Mousavi-Nasab 1 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran AUTHOR R Baharlou 2 Department of Microbiology, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR M Ghaderi 3 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran AUTHOR M Doosti 4 Infectious and Tropical Disease Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran AUTHOR SMA Hashemi 5 Department of Student Research Committee, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR A Samie 6 Department of Student Research Committee, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR S Jahanabadi 7 Infectious and Tropical Disease Research Center, Shahid Sadoughi University of Medical Sciences, Yazd, Iran AUTHOR A Ahmadi-Vasmehjani 8 Department of Microbiology, Jahrom University of Medical Sciences, Jahrom, Iran AUTHOR

ORIGINAL_ARTICLE Background and Aims: Influenza virus nucleoprotein (NP) has the capacity to be used as subunit vaccine, but little is known about the impact of different cultures on its structure. In the present study we aimed to evaluate and compare the Isoelectric focusing (IEF) property of extracted viral nucleoproteins derived from Madin Darby canine kidney (MDCK) cell line and embryonated chicken eggs (ECE). Materials and Methods: Influenza virus strain A/NewCaledonia/20/99/H1N1 was propagated and grown in allantoic sac of 10-11 day-old embryonated chicken eggs, and mammalian cell culture (MDCK) in parallel. Ribonucleoprotein extraction was conducted from two separate cultures and evaluated using isoelectric focusing gel strips. Results: The results showed higher isoelectric pH in extracted nucleoproteins from MDCK as compared to embryonated chicken eggs. Conclusion: It is possible that some amino acids have been replaced. Suggesting that the changing net charge of protein may be affect the conserved regions of the protein. Therefore, this could impact the new generation of vaccines construction based on conserved proteins. http://journal.isv.org.ir/article-1-119-en.pdf 2015-01-11 51 56 10.21859/isv.7.1.2.51 Nucleoprotein MDCK Embryonated chicken egg Isoelectric focusing Yasin Panahi 1 Influenza Unit, Virology Department, Pasteur Institute of Iran, Tehran, Iran AUTHOR Behrokh Farahmand 2 Influenza Unit, Virology Department, Pasteur Institute of Iran, Tehran, Iran AUTHOR Rasoul Soleimani-Stiar 3 Influenza Unit, Virology Department, Pasteur Institute of Iran, Tehran, Iran AUTHOR Reza Saghiri 4 Biochemistry Department, Pasteur Institute of Iran, Tehran, Iran AUTHOR SHima Fattahi 5 Department of Pharmacology, Science and Research Branch, Islamic Azad University, Tehran, Iran AUTHOR Mansoureh Tabatabaeian 6 Influenza Unit, Virology Department, Pasteur Institute of Iran, Tehran, Iran AUTHOR Masoumeh Tavassoti-Kheiri 7 Influenza Unit, Virology Department, Pasteur Institute of Iran, Tehran, Iran AUTHOR

ORIGINAL_ARTICLE Case Report: Recently a 20-day-old layer flock with mortality has been submitted to the PCR Lab. Infectious Bronchitis Virus (IBV) has been detected in the clinical samples. Results: A phylogenetic tree based on a partial S1 gene sequence showed Iranian IBV variant located in LX4-type cluster. This cluster include all QXIBV-type detected in China and European countries. The highest sequence homology (> 99%) was found with that of a group of the Chinese QX but less similarity with European QX-like (>95%) and another Chinese QXIBV (>94%). Conclusion: Comparison of Nucleotides sequence with H120 and 4/91 IB vaccines shows only 76.9 and 79.7 identity, respectively. http://journal.isv.org.ir/article-1-120-en.pdf 2015-01-11 57 60 10.21859/isv.7.1.2.57 Infectious Bronchitis Virus (IBV) Clinical Samples Phylogenetic tree Mohammad Hassan Bozorgmehri-Fard 1 Department of Clinical Science, Faculty of Veterinary Medicine, Islamic Azad University, Science and Research Branch, Tehran, Iran AUTHOR Saied Charkhkar 2 Department of Clinical Science, Faculty of Veterinary Medicine, Islamic Azad University, Science and Research Branch, Tehran, Iran AUTHOR Hossein Hosseini 3 Department of Clinical Sciences, Faculty of Veterinary Medicine, Islamic Azad University, Karaj Branch, Karaj, Iran AUTHOR

ORIGINAL_ARTICLE No abstract this article: A Nested RT-PCR Method for Confirming the Absence of all Three Types of Polioviruses in Primates and Human Cells Used in Vaccine Production http://journal.isv.org.ir/article-1-121-en.pdf 2015-01-11 61 63 10.21859/isv.7.1.2.61 Nested RT-PCR Poliovirus A Salimi-Jeda 1 Department of Human Viral Vaccine, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR A Mohammadi 2 Department of Human Viral Vaccine, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR S Shahmahmoodi 3 Iran National Polio Laboratory, Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran AUTHOR A Foroughi 4 Department of Human Viral Vaccine, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR B Alirezaie 5 Department of Human Viral Vaccine, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR