Application of Real-Time PCR method for evaluation of measles vaccine heat stability

     The Plaque Forming Unit(PFU) and Tissue Culture Infectious Dose50(TCID50) methods are used for evaluation of vaccine heat stability and effect of various stabilizers on thermal stability of vaccines. The aim of present study is using Real-Time PCRtechniquefor estimation of vaccine degradation rate and thermal stability of measles vaccines. Lyophilized measles vaccines containing three various stabilizers were reconstituted with distilled water. Three vial of each vaccine incubated at25˚C for 0, 4 and 8 hours. Titer of virus in vaccines calculated by TCID50 method. Also after RNA Extraction and cDNA synthesis, the RNA copy numbers of viruses in vaccines were estimated by absolute quantitative Real-Time PCRtesting. The data were analyzedby SPSS 19 and Sigma Plot 11 software.The result of this study showed there is a significant relationshipbetween vaccine degradation rate calculated with TCID50 and Real-Time PCR method (p<0.05). ThereforeReal-Time PCR is a good complement or appropriate replacement to traditional methods.Titration methods based on cell culture are gold tests for titration of viral vaccines and estimation of heat stability but Real-Time PCR technique can also be used for this goals. This method is faster, cheaper and easier than TCID50

Increased the specificity and sensitivity of monospecific antibody against host cell protein (HCP) in quality control of hepatitis B recombinant vaccine

     One of the most important aspects in recombinant biologic production, based on GMP rules, is the accuracy of final product quality control, especially assessment of host cell macromolecules contamination rate in final product. The purification requirement can be eliminated when the yeast cell containing the recombinant protein is used as a host cell. It is possibile that the final product contaminated  to the host cell protein during purification stages of HBsAg (HBV vaccine). The protein purification costs depend on the purification  procedures required. Nowadays several companies produce commercial kits for identification and assessment of host cell protein contamination based on ELISA and Western blotting methods. But high prices, difference in sensitivity and lack of easy access to these kits sometimes create problems. So, in this study, two methods of Ammonium sulphate and caprilic acid precipitation technique were used separately for IgG purification. The results showed that IgG purification increased by 97% in caprylic acid method, compared with only a 77% increase in ammonium sulphate method. There were also significant differences in specificity and sensitivity between our standardized ELISA technique and using commercial kit (Cygnus CHO HCP).

In vitro evaluation of inhibitory effect of Lactobacillus reuteri supernatant on the replication of herpes simplex virus type 1 and expression of UL54, UL52 and UL27 genes

Background and Objectives: Human herpes virus type 1 (HSV-1) is a neurotropic pathogen that is infected more than 70% of the world population. The increasing of viral resistance to antiviral drugs and the emergence of side effects has motivated researchers to study the use of probiotics as new antiviral agents. The aim of the present study was to study for the first time the potential antiviral activity of Lactobacillus reuteri (L. reuteri) supernatant against HSV-1. Materials and Methods: After measuring the cytotoxicity of L. reuteri supernatant by MTT assay, 1:16 dilution of it was added to HeLa cells before and after HSV-1 infection, after 1.5 hours incubation with HSV-1, and simultaneously with HSV-1 infection. After 48 hours of incubation at 37°C, the viral titer and expression levels of UL54, UL52 and UL27 genes were measured by tissue culture infectious dose 50 (TCID50) and Real-Time PCR methods, respectively. Results: HSV-1 titer in the treatment conditions before infection, incubation with HSV-1, simultaneously with infection and after infection was reduced by 0.42, 3.42, 1.83, and 0.83 log 10 TCID50/ml, respectively. When the bacterial supernatant was first incubated with the virus and then added to the cell, or when it was added simultaneously with the virus, the expression of the UL27, UL52, and UL54 genes decreased significantly (p0.05). Conclusion: The study findings indicated that the supernatant of L. reuteri has a potent anti-HSV-1 effect especially if it is incubated with the virus before inoculation into the cell. Its possible antiviral mechanism is to inhibit the virus by binding to it or changing the surface structure of the virus. Metabolites of L. reuteri can be considered as a novel inhibitor of HSV-1infection

Human herpesvirus type 6 in patients with systemic lupus erythematosus

Background and Objectives: Infectious agents are considered one of the possible etiological factors of systemic lupus erythematosus (SLE). It has been suggested that human herpesvirus type 6 (HHV-6) may trigger autoimmune disorders, but few studies have been conducted on the relationship between this virus and autoimmune diseases, especially SLE. The present study aimed to compare the frequency of HHV-6 infection between SLE patients and healthy individuals. Materials and Methods: Serum samples were collected from 60 healthy people and 60 SLE patients referred to the rheumatology clinic of Shahid-Beheshti Hospital, Kashan, Iran, from January 2020 to January 2021. The following data were collected from the medical records of patients: sex; age; duration of disease; SLE clinical manifestations; and disease activity. After the extraction of viral DNA from samples, a nested polymerase chain reaction (PCR) test was performed to detect HHV-6. Results: HHV-6 was detected in 12 SLE patients (20%) and in 8 healthy individuals (13.3%). A significant correlation was not obtained between SLE and the presence of HHV-6 (P = 0.09). There was no correlation between musculoskeletal involvements, skin lesions, renal manifestations, and hematological manifestations with the presence of HHV-6 (P˃0.05). HHV-6 was detected more frequently in patients with active lupus than in patients with quiescent disease, but this difference was not significant (P=0.08). Conclusion: Although patients with SLE had a higher prevalence of HHV-6 compared with healthy people, there is no strong link between HHV-6 infection and SLE. Future research is necessary because this data does not support the hypothesis that human herpesvirus 6 plays a role in the pathogenesis of SLE

Survival Outcome in Men with Prostate Cancer in Yazd Province, Central Iran, from 2001 to 2012

Background: Prostate cancer is the second-leading cause of cancer death among men in the world. This cancer has increased significantly during recent years in Iran and is the sixth most common malignancy in Iranian men. Survival rates for prostate cancer in Iran are unknown. The present study aimed to estimate survival of patients suffering from prostate cancer in Yazd, Iran. Methods: In this retrospective cohort study, data were collected from 100 men with prostate cancer registered in Shohadaye Kargar and Shahid Sadoughy hospitals in Yazd during the period 2001-2012. The Kaplan-Meier method was used to estimate the survival over time and Cox regression analysis was performed to calculate the hazard ratios according to demographic and risk variables. Results: The mean age of all patients was 74.4 years. They had a mean survival time of 70.78±4.94 month. The 1-year, 3-year, and 5-year survival rates were 97%, 73%, and 54%, respectively. A statistically significant correlation was found between age, tumor grade, cancer stage, treatment and survival time of patients (p0.05). Conclusion: In this study, 5-year survival of patients with prostate cancer was poor. Screening of men for cancer diagnosis in early stages and use of advanced treatment option, it seems to improve survival outcome. Further studies are needed to monitor the survival of patients with prostate cancer in Iran