Abstract

      Background and Objective: Agr is the most important regulatory system for the expression of Staphylococcus aureus virulence factors in different conditions. Agr acts as a quorum sensing system in this bacterium which is activated by increased cell concentration during the transition from logarithmic growth phase to stationary phase. Its role is to upregulate the secretory virulence factors such as alpha-hemolysin and inhibit the transcription of surface proteins including protein A-encoding gene. The aim of this study was to assess the relationship between the agr system expression and some virulence factors of Staphylococcus aureus in Brain-heart infusion (BHI) culture medium.

     Methods: The expression level of agrA and RNAIII genes from the agr locus along with the expression of hla, spa and mecA genes in BHI broth were assessed in different growth phases using Real time-PCR. Also, gyrB was used as an internal control in this study.

     Results: The growth curve of the five tested isolates in BHI broth at 24 hours showed that all the isolates had relatively similar growth patterns. AgrA gene expression in the stationary phase was decreased by 0.89-fold compared with the logarithmic phase. Although the expression of RNAIII gene increased by 3-fold, hla expression decreased by 0.47-fold.

     Conclusion: An inactive agr system is observed in the BHI broth medium. BHI broth medium contains high amounts of suitable nutrients for the growth of Staphylococcus aureus, thus the bacteria do not require the activity of the agr system for the regulation of the virulence genes in these conditions.

ABSTRACT

       Typing of bacteria is an important part of epidemiological studies on nosocomial infections. Bacterial identification methods have dramatically improved in recent years, which is mainly due to advancements in the field of molecular biotechnology. In many cases, molecular techniques have replaced phenotypic typing methods.

Currently, a wide range of bacterial typing techniques is used that are different from one another in the aspects of study objectives, costs, reliability and discriminatory power. None of the typing methods can achieve all desired objectives of a study alone.

Different typing methods are used for various purposes including: 1. confirmation of epidemiological relationships in spread of an infection, 2. providing epidemiological hypotheses about epidemiological relationships between bacteria in the absence of epidemiological data, 3. describing the distribution of bacterial types and identification of affecting factors. Inferences of epidemiological studies depend on the chosen typing technique and objectives of the study.

Therefore, the typing technique can be useful and effective in increasing our understanding of the pathogenesis, transmission and prevention of possible diseases. The aim of this study was to evaluate various methods of molecular typing of bacteria and to compare these methods from different aspects.

ABSTRACT

        Background and Objective: The current challenge of diabetes mellitus is to prevent its complications. These complications are directly associated with hyperglycemia in diabetics. The HbA1c measurement is essential for long-term glycemic control. Synchronization of HbA1c measurement is important in order to avoid discrepancies between results reported by laboratories. This study aimed to evaluate the accuracy, precision and agreement of five HbA1c measurement methods with HPLC reference method.

       Methods: HbA1c levels of 55 samples were measured using six methods of microcapillary electrophoresis (Sepia), enzymatic method (Pishtaz Teb), immunoturbidometry (Pars Azmoon), boronate affinity (Nycocard), immunofluorescence (ichroma) and Tosoh G8 HPLC.

       Results: The five tested methods showed a good agreement with the HPLC method with correlation coefficient of less than 95%. Regression testing of HPLC method and other methods showed slope of 0.99 (P<0.05) for Sebia, 1.02 (P<0.05) for Pishtaz Teb, 0.79 (P<0.05) for Pars Azmoon, 0.82 (P<0.05) for Nycocard and 0.89 (P<0.05) for ichroma. Average inaccuracy for the Sebia, Pishtaz Teb, Pars Azmoon, Nycocard and ichroma in comparison with the HPLC reference method were -0.09, -0.004, -0.75, -0.79 and -0.78, respectively.

         Conclusion: The Sebia microcapillary method and Pishtaz teb enzymatic method have appropriate accuracy and precision. Therefore, these methods can be used as alternatives to the HPLC method for HbA1c measurement. Other methods such as Pars Azmoon, Nycocard and ichroma have significant shortcomings in terms of accuracy.