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Introduction: A high prevalence of HCV infection among hemodialysis patients has been reported worldwide. Risk factors such as history of blood transfusion, duration of hemodialysis and recently nosocomial transmission of HCV in hemodialysis units have been identified. In this study the prevalence of Hepatitis C virus antibody and risk factors in hemodialysis patients in Markazi province is investigated. Materials and Methods: In this cross-sectional analythical study, blood samples were obtained from all 204 hemodialysis patients. Samples were tested for anti-HCV antibodies by using third generation enzyme immunoassay. The reactive samples on ELISA were confirmed by the third generation RIBA. Risk factors were evaluated by a questionnaire. Data was analysed using Chi square and logistic regression. Results: The prevalence of anti-HCV antibody among hemodialysis patients was 4.9%.Duration of hemodialysis was identified as a major risk factor in transmission of HCV (p=0.004). There was a significant relationship between anti-HCV positivity and previous renal transplantation (p=0.032). Female sex was another risk factor for HCV infection (p=0.030). There was no significant relationship between anti-HCV positivity and history of blood transfusion. Conclusion: Nosocomial transmission of HCV within hemodialysis units seems to be a route of infection in patients on hemodialysis in Markazi province. Application of dialysis precautions recommended by CDC can reduce the prevalence of HCV infection among hemodialysis patients in this province.

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Introduction: Nowadays, the attention of researchers has been focused on natural medicine in order to avoid the detrimental side effects of chemical drugs. In this study we assessed the effect of root extract of Tagetes minuta against HSV-1 and HSV-2. Materials and Methods: This research is an experimental study. Root extract of Tagetes minuta was obtained with 70% ethanol by maceration. Vero cells were grown in DMEM containing 5% fetal bovine serum. Serial dilutions of extracted suspension (1/10, 1/20, 1/40, 1/80, 1/160) were incubated by the exact titer of viruses and monitored for antiviral activity of extract. Data was analyzed using Doncan test. Results: Root extract obtained from Tagetes minuta significantly has antiviral activity against HSV-1 and HSV-2. This extract has more effect on HSV-2 than HSV-1. This study indicates that antiviral activity of the extract varies between different concentrations and the optimum antiviral activity on both viruses was obtained using 1/10 concentration. Conclusion:The results of this investigation showed that root extract of Tagetes minuta have good antiviral potenoial against HSV-1 and HSV-2, a good source of drug for treatment of diseases due to HSV-1 and HSV-2.

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Background: Radiotherapy is one of the cancer treatment methods. Prescribed dose for each fraction is considered based on radiosensitivity of tumoral and normal tissues. Viral agents are the effiectiv factors on tissue sensitivity. This research aimed to determine the effect of ionizing radiation of Cobal 60 on radiosensitivity of Hela cells infected with Measles virus. Methods and Materials: In this study, the radiosensitivity of Hela cells is investigated experimentally and qualitively. The cells have been cultivated in two groups (experimental and blank) and plating efficiency has been obtained. Then 100λ measles virus with serial dilution method was used to induce infection in different ratio for experimental group. After cell growth and passage, the two groups were irradiated with 2Gy gamma radiation of cobalt 60. Results: Results respectively indicated cell death increases up to 5-7%, 15-20% and 50-65%, after 2Gy irradiation by Co 60 for contaminating to Measles in low, moderate and high concentrations. Conclusion: Radiosensitivity of tumoral cells increases when they are infected by viral agent. The result in radiotherapy of cancers showed, in prescribing dose fraction non cancer disease should be considered.

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Background: Multiple sclerosis (MS) is a auto-immune disease of central nervous system. The etiology of MS is unknown, but environmental factors such as viruses are involved in the development of MS. In this study, MS patients were assessed for antibodie titers against Human Herpes virus-6 (HHV-6) in Markazi Province. Methods and Materials: In this case-control study, 31 new cases of MS patients and 60 healthy subjects were selected with similar demographic criteria such as sex, age and location. Antibodies titer (IgM and IgG) against HHV-6 were examined by ELISA and Immunofluorescence methods. Data were analyzed using Logistic regression and Odds ratio. Results: Data indicates that 74.2% of case group and 34.2% of control group were identified as positive for IgM against HHV-6. The difference between the two groups in terms of IgM against HHV-6 was statistically significant (p=0.001). Incidence of IgM positivity against HHV-6 was increased more than five times in MS patients compared to control group. Also there was a statistically significant difference between case and control groups in IgG titer (p=0.019). Conclusion: Acute infection of HHV-6 is a risk factor for MS.

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Background: Herpes simplex clinical manifestations are in the form of vesicular eruptions on erythemateus base. The virus can remain latent within sensory nerve ganglions following the initial infection and be reactivated in some cases such as intracranial surgery, dental surgery, stress and excitements. Systemic or disseminated infection occurs in immune-deficient patients and sometimes in healthy individuals. In Previous studies, in most of the disseminated infection cases, visceral organs involvement has been mostly observed while skin involvement has been reported just in one case. Thus, our patient was the second case with herpes simplex disseminated skin infection. Case: The patient was a 38 year-old man who underwent craniotomy surgery due to epidural hematoma following car accident and head trauma. Almost 10 days after the surgery, the patient developed disseminated vesicular eruptions in erythemathous base which initially appeared in the face, head, and then in body and extremities. Fever was detected 5 days before the development of eruptions. Other than anemia and leucocytosis, there were no other notable points in the systemic examination and lab tests. For patient skin biopsy was taken with an impression of herpes simplex, mulocum contagiosum and a lower probability for bullouse diseases. In the skin biopsy, intra-epidermal vesicles and extensive epidermal necrosis with multinucleated giant cells and intra-nuclear objects were observed. A plethora of neutrophilic cells inside the vesicles and inflammatory cell infiltration in the underlying debris were seen. Based on the pathology report, infection with herpes simplex hominis was confirmed. Conclusion: Disseminated skin infection with herpes simplex can be observed even in healthy people with no background.

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Background: Based on the severity and prognostic condition of respective cancers caused by them, papilloma viruses are classified into high, medium, and low risk groups using E6 and E7 viral proteins. Nowadays, different methods of modeling in clinical medicine are used for diagnosis of diseases and evaluation of their molecular characteristics. Among the new methods of modeling, fuzzy systems are of particular importance in various fields of science. The aim of this study was to use a new intelligent Adaptive Nero Fuzzy Inference System (ANFIS) for predicting human papilloma virus oncogenicity based on a number of biochemical properties of E7 protein. Materials and Methods: In this study, using ANFIS model, a new model was developed for predicting oncogenicity of papilloma virus isolated from patients. The process of training and testing was performed using a set of available published filed data and several statistical and graphical criteria. Accordingly, through provision of needed biochemical and biophysical data on E7 gens from the existing data, this model was developed. The results of this model were, then, validated by the authentic published data. Results: Based on the results, the developed model is capable of predicting papilloma virus oncogenicity efficiently. R2 and RMSE values in training stage were 0.99 and 101.18, respectively. In the testing stage, however, they stood at 0.94 and 173.8, respectively. Conclusion: Based on the findings, the use of ANFIS model significantly improves the accuracy of estimating virus oncogenicity phenomenon. The methodology presented in this study is a new approach in estimating viral oncogenicity and can successfully be combined with other mathematical models for model updating in real conditions.

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Background: TTV is the first human circoviridae that was isolated from Japanese patients with unknown hepatitis in 1997. Since then, several studies have been done on different aspects of TTV pathogenesis. The aim of this study is to determine the prevalence of TTV in patients with chronic hepatitis using two different primer sets. Materials and Methods: In this descriptive study, blood samples from 240 patients with chronic hepatitis C at Professor Alborzi Clinical Microbiology Research Center were assessed in terms of the presence of TTV DNA in plasma through the nested polymerase chain reaction using two primer sets. Results: Of the 240 patients, TTV-DNA was detected in 220 (92%) patients with chronic hepatitis C using 5΄-UTR primer and in 12 (5%) patients using N22 primer. According to the demographic data, there was not a significant difference between male female patients in prevalence of TTV infection. Conclusion: The prevalence of TTV DNA in plasma samples from patients with chronic HCV by using 5΄-UTR primer was high and it was congruent with studies done in other countries however, N22 primer showed a lower prevalence of viral DNA in the samples. Overall, there was not a significant correlation between sex and the presence of viral DNA in patients. Controversial or high prevalence of this virus in HCV infected people necessitate further studies for determining the relationship between HCV and TTV infection.

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Background: Several different molecular methods have been developed that are capable of detecting HIV-1 in clinical specimens with different levels of sensitivity and specificity. This article describes the results of a reliability study on the development and application of a new real-time TMA method for isothermal detection of HIV-1. Materials and Methods: In this ex Primental study, the molecular beacon primer and probe set were designed for a 176-base-pair region of HIV-1 pol gene using a specialized software. Logarithmic serial dilutions from 10-107 copies of an in-vitro transcribed RNA were used for determination of the analytical sensitivity of the assay. Clinical specimens that had previously been evaluated positive or negative by a valid commercial assay were used for assessing the clinical sensitivity and specificity of the assay. Results: The analytical and clinical sensitivities of the assay were determined 500 copies/ml and 93.3%, respectively. The primers and the probe were HIV-1 specific and no cross-reaction was observed with other blood-borne viruses and human genome bioinformatically. The clinical specificity of the developed real-time TMA assay was examined experimentally using 20 negative samples and determined to be 100%. Conclusion: The developed real-time TMA assay can be used as an appropriate tool for the rapid and isothermal detection of HIV-1 in patients' blood and plasma samples.

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Background: Noticing the sensitivity of measles virus to temperature and light, maintaining its stability is highly important in live vaccines. The aim of the study is to evaluate the stability of measles vaccine produced by AIK-C strain. Materials and Methods: In this experimental study, three lyophilized vaccine vials were incubated at 37˚C for one week and their stability was evaluated via accelerated test. In addition, reconstituted vaccines were incubated at 4˚C, 25˚C, and 37˚C for 0, 4, 8, 12, 16 hours after reconstitution and their remaining infectious virus titer was measured using CCID50 method. Half-life of the reconstituted measles vaccine was evaluated according to linear regression analysis. Results: When the reconstituted vaccine was incubated at 4˚C, 25˚C, and 37˚C, the titer loss per hour was equal to 0.05, 0.1 and 0.2 Log10 CCID50, respectively. Also, the half-life of this vaccine at these temperatures was 5.31, 2.26, and 1.36 hours, respectively. Conclusion: The loss of potency for measles vaccine produced by AIK-C strain is 0.33 Log after storage at 37°C for one week, while the reported amounts for commercial vaccines such as Mevilin-L, Attenuvax, Edmonston-Zagreb, and Rimevax are 0.7, 0.7, 1 and 0.78, respectively. Lyophilized and reconstituted vaccine containing AIK-C strain is more stable in comparison with Edmonston B, Schwartz, Biken-CAM, and Leningrad strains. The stability of the reconstituted AIK-C strain vaccine is similar to Moraten strain at 37˚C.

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Background: In recent years Influenza viruses have caused widely spread moderate to severe infection in all around the world and there is no Influenza vaccine which can protect people only with one dose injection till now. Therefore , producing a universal vaccine based on virus like particle (VLP) could be ideal. In this study one of the molecular structures was considered for VLP based Influenza vaccine. Materials and Methods: In this experimental study, the human influenza virus (A /New Caledonia 20/1999/ (H1N1)) was propagated in MDCK cell culture. Viral RNA was extracted using RNX-plus solution. Complementary DNA synthesis was carried out using uni-12 primer and random hexamer as specific and general primers, respectively. Neuraminidase open reading frame (1413-bp) was amplified by PCR and cloned into pBlue-script SK. Neuraminidase coding frame sub cloned into pFastBac11 plasmid through SalI/XhoI sites. After verification of cloned Neuraminidase by restriction analysis, it was subjected to automated sequencing bi-directionally. The recombinant pFastBac Neuraminidase vector was transformed to E.coli DH10Bac cells which harbor bacmid DNA and helper plasmid to create Neuraminidase recombinant bacmid. Results: Neuraminidase recombinant bacmid was created by homologous recombination between pFastBacNA and bacmid and was verified by PCR using Neuraminidase specific and M13 universal primers. Conclusion: Recombinant baculovirus expressing Neuraminidase gene can be also used with other individual recombinant baculoviruses expressing HA and M1 genes in production of influenza VLPs or proteins resulting from this structure could be purified in specific insects for vaccine research studies.

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Background: Chronic hepatitis B virus (HBV) infection is a multi-factorial disease that is accompanied with serious clinical complications. Host’s genetic background, especially immune–genetic factors, is critical in the pathogenesis of infection. Gamma interferon ((INF-γ) and its receptor have an important role in immune response to the virus and clinical course of the disease. The aim of this study is to investigate the association between single nucleotide polymorphism -611G/A located in promoter of gamma interferon receptor1 gene (INFGR1) and chronic HBV infection. Materials and Methods: In this Case Control study, genomic DNA from peripheral blood samples of 150 chronically HBV infected patients and 150 healthy controls was extracted by phenol-chloroform method. DNA analysis was performed by PCR-RFLP method and P<0.05 was considered significant. Results: After stages of genotyping and statistical analysis, a significant difference was observed between patient and control group, so that genotype GG was higher in the control group compared to the patient group. Conclusion: The host’s immune-genetic background can play an important role in the pathogenesis of infectious disease. Variations in INFGR1 were related to several diseases. The results showed that the presence of GG allele is accompanied by a decrease in susceptibility to chronic HBV infection.

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Background: Genetic variability of influenza viruses causes new epidemics worldwide annually. Development of a new vaccine for prophylaxis of influenza virus has been amajor objective in recent years. The aim of this study was to construct a recombinant baculoviruscapable of expressing the two surficial antigenic glycoproteins, hemagglutininand neuraminidase, as well as matrix proteinsof swine influenza (H1N1) simultaneously and independently. Materials and Methods: In this experimental study, first, a triplet cassette providing simultaneous and independent expression of target proteins was designed and subjected to synthesis. It was then cloned into pFastBac1 donor plasmid. Competent E.ColiDH10Bac cells were transformed by donor clone and the recombinant bacmids were produced following homologous transposition. This construction was verified by PCR and then transfected into Sf9 insect cells to package new recombinant baculoviruses. Results: Restriction map of pFastBacI HNM1 donor plasmid confirmed the fidelity of the clone. The results of PCR done on the recombinant bacmidas template indicated that a proper homologous recombination has occurred between pFastBacI HNM1 donor plasmid and the bacmid in E.ColiDH10Bac host cells. Protein analysis of the infected Sf9 cells showed that all target proteins were efficiently expressed at the same time. Conclusion: The recombinant baculovirus constructed in this studypossesses proper characteristics to produce swine influenza virus-like particles in Sf9 cells.

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Background: Influenza type A virus is one of the most important viral agents in human respiratory diseases. The genetic variability of the influenza viruses leads to the incidence of new epidemics worldwide. Hence, there is a growing need for rapid and effective new methods capable of detection and differentiation of influenza virus circulating strains. This study was done to develop a method for rapid differentiation of the subtypes of influenza type A virus. Materials and Methods: In this experimental study, reverse-transcription and polymerase chain reaction (RT-PCR) were performed using a primer set based on M gene of H1N1, H3N2, H5N1, and H9N2 influenza subtypes. Then the amplified fragments were subjected to digestion using subtype specific restriction endonuclease enzymes. Results: The results of PCR reaction showed that the primer pair of the M gene was specific and capable of amplifying all influenza subtypes understudy. Also, different restriction fragment length polymorphism patterns (RFLP) were generated using enzyme digestion reaction on the amplified segment of M gene. Conclusion: RT-PCR and RFLP analysis of the M gene can be employed as a useful method for differentiating influenza virus subtypes

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Background: The importance of VP2 protein of canine parvovirus to bind to human cancer cells and to detect the virus in veterinary detection kits has motivated a lot of research on the production of this protein. In this project, a surface gene of canine parvovirus (VP2) was cloned and expressed in a prokaryotic vector system and its expression was optimized in a specific cell-free prokaryotic expression system. Materials and Methods: In this experimental study, plasmid pET-21aVP2 was constructed by cloning the PCR product of VP2 gene of canine parvovirus into the plasmid expression pET-21a vector. The best sequence was analyzed through PCR and it was followed by confirmation with sequencing and restriction digestion. To produce VP2 protein, plasmid pET-21aVP2 was transferred into Escherichia coli, Rosetta (DE3) strain, and the expression of this protein was induced by IPTG. The production of VP2 protein in both systems was evaluated using SDS PAGE technique. The expressed protein was checked with monoclonal antibody against VP2 protein by Western blotting technique. Results: Successful cloning of VP2 protein was confirmed by enzymatic digestion and sequencing. The expression of VP2 protein in bacterial and cell-free prokaryotic systems was verified by SDS PAGE and the specific band in Western blotting also confirmed the VP2 protein. Conclusion: The results of this study showed that VP2 gene was amplified in the cloning phases and it was successfully cloned in the expression vector. Protein expression was confirmed in both bacterial and cell-free prokaryotic systems.

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Background: The Real-time PCR assay has been established as the standard method for Human Cytomegalovirus (HCMV) quantitation in immunocompromised patients. However, the question of which one of whole blood or plasma specimens is better for viral quantitation is still unresolved for many clinical laboratories. To answer this question, the current study compares HCMV DNA load in whole blood and plasma samples.

Materials and Methods: In this prospective study, the whole blood and plasma samples were obtained from 41 transplantated patients and the viral load was detected using a validated, in-house Real-time PCR assay.

Results: Of the total 193 examined specimens, 174 were negative and 19 samples, from 16 patients, were positive in at least one of whole blood or plasma samples. Based on the results of linear regression analysis, the cytomegalovirus viral load was correlated in whole blood and plasma samples (R2: 0.872). However, the regression equation shows that the HCMV load in whole blood samples is higher than load of this virus in plasma. The validity of the quantitative results was confirmed by repeating the tests and analyzing the results using the repeated measure analysis.

Conclusion: Based on the results of the present study, HCMV quantitation in whole blood samples has a higher analytical sensitivity than in plasma samples.

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Background: Newcastle disease virus (NDV) is one of the major pathogens in poultry and vaccination is intended to control the disease, as an effective solution, yet. Fusion protein (F) on surface of NDV, has a fundamental role in virus pathogenicity and can induce protective immunity, alone. With this background, here our aim was to construct a baculovirus derived recombinant bacmid shuttle vector (encoding F-protein) in order to express it in insect cell line.

Materials and Methods: In this experimental study, at first complete F gene from avirulent strain La Sota of NDV was amplified by RT-PCR to produce F cDNA. The amplicon was cloned into T/A cloning vector and afterwards into pFastBac Dual donor plasmid. After the verification of cloning process by two methods, PCR and enzymatic digestion analysis, the accuracy of F gene sequence was confirmed by sequencing. Finally, F-containing recombinant bacmid was subsequently generated in DH10Bac cell and the construct production was confirmed by a special PCR panel, using F specific primers and M13 universal primers.

Results: Analysis of confirmatory tests showed that the recombinant bacmid, expressing of F-protein gene in correct sequence and framework, has been constructed successfully.

Conclusion: The product of this F-containing recombinant bacmid, in addition to its independent application in the induction of protective immunity, can be used with the other individual recombinant baculoviruses, expressing HN and NP genes to produce NDV-VLPs in insect cell line.

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Background: Because of the reported high ability of virulence and medicinal resistance of HIV-1 virus during the last decades, many investigations have been performed concerning discovery and the introduction of anti-HIV-1 drugs. The results of numerous researches have shown that drugs and protease inhibitory compounds mainly containing plant derivatives specially terpenoids may control HIV-1 infection very effectively. The aim of this research is the bioinformatical study of HIV-1 protease inhibition by standard drugs and triterpenoides from plant and mushroom.

Materials and Methods: This is a descriptive-analytic study. In the present study , the structure of drugs, triterpene comounds, and HIV-1 protease enzyme was received from the databases such as Chem Spider, PubChem, Human Metabolome Database (HMDB), and Protein Data Bank (PDB). After that, molecular docking was performed by iGRMDOCK 2.1 software

Results: The results confirmed that the interactions of the triterpene compounds like the standard drugs were in three safeguarded and catalytic areas including central domain, flap and carboxylic terminal domain specially amino acids Asp25, Asp27, Ala28, Asp29 and Asp30 in active sites of HIV-1 protease. Also, The study of the interactions of these areas showed that there is a direct correlation between the strength of the interactions and IC50 values of these compounds.

Conclusion: Finally, with due attention to the high effectiveness and the proprietary function of triterpenoids, we can conclude that these compounds may be considered as effectire HIV-1 antiprotease drugs.

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Background: In 1970, human papillomavirus (HPV) was introduced as the main etiologic factor of cervical carcinoma. Since there is no possibility of detecting the virus and its subtypes using serological methods and cell culture, the molecular methods such as PCR have particular importance in accurate, early and definite diagnosis of the virus. So, in this research, our goal is to use a proprietary PCR assay based on L1 gene of human papillomavirus for molecular recognition of HPV and to evaluate its prevalence in patient samples.

Materials and Methods: In this experimental study, after collecting of samples from malignant cervical lesions, the viral DNA was extracted from paraffin blocks of 50 clinical samples and PCR was done by specific primers for L1 gene of human papillomavirus in all samples. After the analysis of PCR products by 2% agarose gel electrophoresis, sensitivity and specificity of the test were also evaluated.

Results: Among 50 patient samples, 33 cases were confirmed to be positive for HPV infection and 17 cases were negative, showing high frequency of HPV in this patient population (about 66%). The results of specificity assay were positive for papilloma samples and sensitivity of the assay was 20 copies of recombinant construct containing L1 per reaction.

Conclusion: This study showed that PCR by specific primers for L1 gene of human papilloma virus is a proper and accurate method for detection of this virus and the results confirm the previous reports of correlation between HPV and cervical carcinoma.

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Background: Direct transmission of avian influenza viruses with human receptor binding specificity to humans is a serious risk of newly emerging virus responsible for pandemy. The analysis of recent avian influenza hemagglutinin sequences and their glycans show their affinities to the human sialic acid receptors. The upregulation of proinflammatory cytokines and type I IFN genes, and host cell death responses contribute to the pathogenesis of influenza infection. Understanding the host cell-virus interactions and replication dynamic of the viruses in different cells is an essential step in surveillance and controlling programs against influenza.

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Background: Hepatitis B virus (HBV) is a member of hepadenaviridae family, which is infectious for humans and a few animal species. Successful clearance and elimination of infection from the body or development of HBV infection to chronic disease depend on the host genetic background in immune system genes. Interleukin-12 (IL12) and also Interleukin-12 Receptor B1 (IL 12 RB1) are the key factors in the spontaneous clearance of viral infections, especially HBV. The aim of the present research is to investigate the association between Interleukin-12 receptor B1 gene polymorphism (rs11575934 A/G) and susceptibility to chronic Hepatitis B virus infection.

Materials and Methods: In this case-control study, genomic DNA of 150 chronic HBV infected patients and 150 healthy controls were extracted from peripheral blood cells. Single nucleotide polymorphism (rs11575934 A/G) was genotyped using polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP).

Results: The frequency of GG, AG, AA genotypes was 6.7%, 40.7%, and 52.7% in chronic patients and 12.7%, 41.3%, and 46% in control group, respectively. No statistically significant difference between case and control groups has been observed (p=0.176).

Conclusion: In the present study, no significant correlation between rs11575934 A/G single nucleotide polymorphism of the IL12RB1 gene and susceptibility to chronic hepatitis B virus infection has been observed. According to the study, this polymorphism does not affect the susceptibility to chronic HBV infection.