Razi Journal of Medical Sciences

fa جداسازی مولکولی ژن Lux سراشیا مارسنس جدا شده از منابع بالینی و تاثیر سوپرناتانت پروبیوتیک لاکتوباسیلوس رامنوسوس بر بیان آن با روش Real time- PCR Molecular segregation of the Lux gene by Serratia marsensis separated from clinical sources and the effect of Lactobacillus Ramenusus probiotic supernatant on its expression by Real-time PCR method علوم آزمایشگاهی Laboratory Sciences پژوهشي Research زمینه و هدف: سراشیا (Serratia)باکتری گرم منفی و متحرکی است که از عفونت‌های ریوی-ادراری و خون میباشد. از خصوصیت پیگمان زایی سراشیا مارسنس به عنوان مارکر ذرات غبار در محیط و در بیمارستان استفاده می‌شود. ژن Lux در اغلب باکتری ها نقش اساسی در پدیده پیام رسانی و در تنظیم بیان ژن نقش کلیدی دارد. این پژوهش جداسازی مولکولی ژن Luxs سراشیا مارسنس جدا شده از منابع بالینی و تاثیر سوپرناتانت پروبیوتیک لاکتوباسیلوس رامنوسوس بر بیان آن با روش Real-time PCR می باشد. روش کار: از بین 100 نمونه از منابع بالینی بیماران بستری دارای عفونت ادراری در بیمارستان‌های مختلف اراک جداسازی و به صورت استریل به آزمایشگاه میکروب شناسی منتقل شد. پس از تایید باکتری توسط تست‌های اختصاصی و جداسازی ایزوله‌های واجد ژن lux استخراج DNA صورت گرفت. تاثیر پروبیوتیک لاکتوباسیلوس رامنوسوس در غلظت های مختلف برای هر نمونه مورد بررسی قرار گرفت. در همه مراحل از یک باکتری سراشیا مارسنس به عنوان کنترل مثبت استفاده شد. یافته‌ها: از تعداد 100 نمونه مورد بررسی 12 باکتری سراشیا مارسنس به دست آمد. 12 جدایه سراشیا مارسنس جداسازی شده همگی (100%) واجد ژن بیوفیلم Lux هستند. میزان Fold Change برای ژن LUXI برابر 09/1- بیانگر آنست که این ژن در گروه تیمار شده با لاکتوباسیلوس رامنوسوس نسبت به گروه غیر تیمار شده 09/1 برابر کاهش یافته است. نتیجه‌گیری: تشکیل بیوفیلم، باکتریها را از فاگوسیتها و مولکولهای سمی حفاظت میکند. باکتریهای حامل ژن lux مولد بیوفیلم در سراشیا مارسنس، تحمل بیشتری نسبت به آنتیبیوتیکها رادارد. با استفاده از پروبیوتیک‌ها و کاهش بیان ژن‌های دخیل در ایجاد بیوفیلم، می توان در درمان شاهد موفقیت های بیشتری استفاده کرد.   Background & Aims: Serratia marcescens is a well-known species of Ceratia, a gram-negative, motile bacterium that is of great clinical importance and grows well in the laboratory. This bacterium is found in natural environments including soil, climate, on the surface of parts of plants and also as an opportunistic pathogen in humans. Sarachiae have small capsules and their colonies are white-pink or red pigment. These bacteria can be isolated from pulmonary-urinary tract and bloodstream infections. Cultivating them smells like fish or urine. Serratia marcescens is a well-known species that is of great clinical importance (1). Biofilm formation is formed by intercellular interactions between bacteria that form the Quorum sensing system. Chromium sensing system is a concentration-dependent process that exists in both gram-positive and gram-negative bacteria. In this system, bacteria communicate with each other through small molecules called self-inducing molecules. When the bacterial density in the environment reaches a certain level, the concentration of these transport molecules reaches a certain level and induce large changes in the level of gene expression. The Qs chromosensing system is governed by the homologs SWrR, SwrI, LuxR, and LuxI, respectively, and affects the expression of at least 28 proteins (3). Probiotics are known as dietary supplements containing live microbes that produce beneficial effects on the host body through the balance of intestinal microflora. The beneficial effects of probiotics through intestinal microflora are known. Probiotics are living microorganisms that will have beneficial health effects on the host in sufficient quantities and include species of Lactobacillus, Bifidobacterium, Bacillus, Enterococcus and Streptococcus (4). The genus Lactobacillus are the most abundant microorganisms that can act as probiotics in the human body. This group of bacteria improves and enhances the function of the immune system, and probiotics, including Lactobacillus, increase the body's resistance to infections and cancers. Consumption of these bacteria increases the function of macrophages and the secretion of various substances, including immunoglobulins (5). The present study was performed to investigate the antimicrobial and anti-binding effects of probiotic Lactobacillus on Serachia marsens and possibly to introduce this bacterium as an inhibitor in the prevention and treatment of urinary tract infections.  Methods: This is a descriptive cross-sectional study that was conducted in 1997. Among 100 clinical sources, patients with urinary tract infections hospitalized in different hospitals of Arak were isolated and sterilized and transferred to the microbiology laboratory. Skim milk agar and biochemical tests were used to isolate and identify microorganisms with proteolytic activity. Using specific primers, Serachia marsense was identified, and the ability to produce the specific pigment Geocin, which is specific to this species, was investigated. Based on macro and microscopic morphology and growth in specific environments, Serachia marcens isolate was obtained. The isolated strains were then stored in TSA media at 4 ° C and glycerinated TSB at -20 ° C until the experiment (7). Results: From 100 samples collected and studied, 12 bacteria of Ceracia martens were obtained. The results showed more biofilm in the strains carrying the lux gene, so the QS luxS gene could affect the initial biofilm formation by the mutant strain. In the present study, by examining the presence of lux gene in patients with clinical infections, its effect on pathogenicity was investigated. Lactobacillus rhamnosus was 1.09 times less than the untreated group, so the results of this study can be attributed to a wider range of probiotic performance, with a review of a wider range of probiotic strains By examining them further, we can identify the power of the effect of different strains of them. Conclusion: According to the results of the present study, lactobacilli have an inhibitory role against many bacteria and the solution can be used in concentrated form, in which case, by increasing the effective compounds in the environment, possibly antibacterial activity. will increase. Methods such as chromatography can also be used to identify effective compounds secreted by these bacteria so that, if possible, by purifying and concentrating them, an effective biological solution to the application of chemicals and increasing resistance can be obtained. It provided a drug in bacteria, especially pathogenic strains.  Biofilm formation protects bacteria, including phagocytes and toxic molecules. Biofilm-producing lux gene carriers, such as Ceracia martensis, are more tolerant of antibiotics, which is an obstacle to their treatment. Probiotic bacteria have the ability to accumulate cells with complex pathogenic microbes. With their anti-binding effects, they prevent pathogenic bacteria from reaching and attaching to the target cell in their host. Therefore, the present study was performed to investigate the antimicrobial and anti-binding effects of probiotic Lactobacillus on the uropathogenic bacterium Ceracia martensis and possibly to introduce these bacteria in the prevention and treatment of urinary tract infections. According to the results of the present study, by using probiotics and reducing the expression of genes involved in biofilm formation, we can see more success in the treatment process and use less common antibiotics.   سراشیا مارسنس,ژن lux,RealTimePCR, لاکتوباسیلوس رامنوسوس Seracia marsensis, lux gene, Real Time PCR, Lactobacillus rhamnosus 25 35 http://rjms.iums.ac.ir/browse.php?a_code=A-10-5075-2&slc_lang=fa&sid=1 Zahra Rozbehanii زهرا روزبهانی 3900319475328460055372 3900319475328460055372 No , Islamic Azad University, Arak, Iran دانشگاه آزاد اسلامی، واحد اراک، اراک، ایران Behroz Shojaii saedii بهروز شجاعی سعدی bhshojaii@gmail.com 3900319475328460055370 3900319475328460055370 Yes Islamic Azad University, Arak, Iran دانشگاه آزاد اسلامی، واحد اراک، اراک، ایران Kumarss Amini کیومرث امینی dr_kumarss_amini@yahoo.com 3900319475328460055371 3900319475328460055371 No Islamic Azad University, Saveh, Iran دانشگاه آزاد اسلامی، ساوه، ایران