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Background and Objective: HIV-1 and HCV infections especially in co-infected forms are among the most important infections transferred during blood transfusion.The screening of the blood products is valuable for preventing the transmission of infections. The aim of this study was to evaluate multiplex RT-PCR assay for detection of Co-infection HIV-1 and HCV Viruses in plasma samples.

Materials and Methods: This laboratory study was done to evaluate the use of multiplex RT-PCR assay for simultaneous detection of HIV-1 and HCV genomes in plasma samples. The amplified genomes were detectable in 3% agarose gel base on difference in the numbers of nucleotides. The sensitivity and specificity of this assay was determined on healthy and infected subjects whome simultanously exhibit HIV-1 and HCV co-infection using plasma samples.

Results: The specificity results showed that the primers used in this assay have no interaction with each other and other possible interfering agents. The clinical sensitivity and specificity of the assay has been considered as 90% and 100%, respectively.

Conclusion: Multiplex RT-PCR can be used for screening of blood donors due to high sensivity and specificity.

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Background and Objective: Continuous antigenic variation of Influenza a viruses causes a major concern to develop Influenza vaccine. Conserved antigens are suitable candidates for vaccine production due to its non-requirement to match the designed strains with circulating strains. The M2 gene is conserved among Influenza a viruses and has potential to be considered as a universal vaccine. This study was designed to evaluate the effects of aqueous Echinacea purpurea extract on immunogenicity of DNA vaccine encoding M2 gene of Influenza virus. Materials and Methods: This interventional study was carried out on female BALB/c mice with 3-4 week age (250-300 gr). Plasmid DNA encoding M2 gene (pcDNA-M2) of Influenza virus A/New Caledonia/20/99 (H1N1) was transformed into E.coli top10 f' and cultured in LB broth media. Large scale plasmid preparation was done and the concentration was measured by spectrophotometric method. Mice were divided into eight groups and immunized three times with fifteen days apart. Vaccine groups received inactivated Influenza virus or pcDNA-M2, alone or in combination with Echinacea extract. Control groups were injected pcDNA, Echinacea extract, and phosphate buffer. All animals were left to bleed before immunization and at 21 days after the last vaccination and specific anti-M2 antibodies were measured by indirect ELISA. Then the mice were intranasally challenged under an aesthesia with mouse-adapted PR8 Influenza virus and monitored for 3 weeks to evaluate the vaccine regimen efficacy in reduction of mortality rate compared to control groups. Data were analyzed using SPSS-16, One-way ANOVA and Kaplan–Meier tests. Results: The highest specific immune response was obtained in mice received inactivated virus plus extract (P<0.05). Immune responses in mice inoculated with pcDNA-M2 were significantly higher compared to all control groups mice (P<0.05). In addition the specific immune responses in group inoculated with pcDNA-M2 and aqueous extract was higher compared to the group receiving only pcDNA-M2 (P<0.001). The highest survival rate was observed in mice injected with inactivated virus or pcDNA-M2 plus extract. Conclusion: This study showed that pcDNA-M2 induced specific immunity and protected mice against lethal challenge with PR8 Influenza virus. Furthermore, application of Echinacea extract with M2 gene vaccine increased vaccine efficacy.

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Background and Objective: Rotaviruses are the members of the Reoviridae family containing double-stranded RNA (dsRNA) genome which are the main cause of gastroinentritis particularly in children less than three years. This study was designed to evaluate the detection of rotavirus genome by new silver staining method using polyacrylamide gel electrophoresis (PAGE). Method: In this descriptive study, the samples were collected from infected MA-104 cell culture and the RNA electrophoresis was performed in 10% polyacrylamide slab gels after RNA extraction. Results: According to polyacrylamide gel electrophoresis and sensitive staining analysis, rotavirus RNA segments were divided into 4 groups and single-nucleotides differences were clearly detected rapidly. Conclusion: New silver staining method using polyacrylamide gel electrophoresis has the capacity to detect the rotavirus electeropherotype within a few minutes even in small DNA/RNA pieces up to 7 picograms.

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Background and Objective: Sea cucumber (Holothuria leucospilota) is used for food purposes and traditional medicine in the South East and East Asia. This study was done to determine the antiviral effect of methanolic extract, of Holothuria leucospilota species against HIV-1 virus. Methods: In this laboratory study, sea cucumbers were collected from Larak Island, Persian Gulf, Iran at depths of 10-30 m. Methanol solvent was used for extraction process. Extract was concentrated by rotary evaporator at 40-45 degree C, and subsequently was prepared in the form of dry powder using vacuum freeze dryer lyophilization. Results: The extract in 100 and 1000 µg/ml of concentrations inhibited by 94% and 92.5% the replication of HIV-1, respectively. 10 µg/ml of extract had not specific antiviral effect. Approximately the half of concentration of extract (35.89 µg/ml) prevents 50% of proliferation of HIV-1, which was 50% toxic of on host cells (P<0.05). Conclusion: Sea cucumber methanolic body wall extract of Holothuria leucospilota species had no antiviral effect against HIV-1 virus. It can be due to cytotoxic effect of extract on the host cells.

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Background and Objective: Chronic infection with Hepatitis B virus (HBV) is one of the main causes of cirrhosis and hepatocellular carcinoma (HCC). The pathogenicity of the virus is determined by the multi-functional protein x (HBx). Changing the sequence of the gene encoding this protein causes the regulation of transcription and pathogenicity factors. This study was done to analyze the genetic dynamics of the HBx coding gene in a person with chronic HBV.
Methods: In this descriptive laboratory study, an infected person with chronic hepatitis B virus infection was first amplified and cloned into complete sequence of HBx encoder. Then, the reference sequences of genotypes, serotypes and different virus subtypes of the GenBank database were matched by CLC Sequence Viewer software. The comparative result was used to plot the phylogenic tree by T-rex server and population genetic analysis using DnaSP software. Natural selection at the nucleotide and protein level was performed by the Tajima's D test.
Results: No known mutation at the level of the protein was found in the chronic sequence of the HBx encoder. The results of natural selection indicated neutral mutations in the HBx gene. The phylogenetic results showed that the HBx encoding sequences in the chronic infected individual had a genetic affinity with genotype D and ayw2 subtype.
Conclusion: Neutrality polymorphism takes place in HBx coding region. Also, the phylogenetic results of the present study are consistent with the previous findings of Golestan province and Iran which have reported the prevalence of genotype D and subspecies ayw2.

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Background and Objective: Since the onset of the COVID-19 (Corona Virus Disease 2019) pandemic, several challenges have been proposed to the disease and the causing viral agent. Accurate and rapid diagnosis of the virus is essential to control the spread and progression of the disease. Choosing a suitable sample in different phases of the disease will reduce the false-negative results. This study was performed to identify the SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) genome in the blood samples of COVID-19 patients.
Methods: This descriptive-analytical study was performed by census method on 100 whole blood samples of patients (50 recovery and 50 deceased) with a definitive diagnosis of COVID-19 (positive Real-Time RT-PCR test of nasopharyngeal swab samples) admitted to Shahid Sayyad Shirazi educational and medical center in Gorgan during 2020-21. Clinical and laboratory findings were compared in the two groups of patients. The viral nucleic acid was extracted from the whole blood samples of the patients, and the presence of the virus genome was investigated using primer and probes via the Real-Time RT-PCR method.
Results: The age of the recovered patients (49.06±15.1 years) was significantly was lower than deceased patients (58.3±12.4 years) (P<0.05). Clinical symptoms including cough, shortness of breath, sputum secretion, and vomiting in deceased patients were significantly more than recovery group (P<0.05). The lymphocytes count and platelet level in the deceased group were lower than in the recovered group. Level of lactate dehydrogenase (LDH) was higher in the deceased group in compare to recovered group (P<0.05). The virus genome identified in the blood samples of 7 patients (3 recovered and 4 deceased), which had no significant relationship with the outcome of the disease.
Conclusion: The use of blood samples for the diagnosis of COVID-19 is not appropriate.