other بررسی قدرت اتصال و حساسیت دو آپتامر اختصاصی جهت تشخیص E. coli O157:H7 با استفاده از روش ELISA سلولی و مولکولی Cellular and molecular مقاله پژوهشی Research Article <span style="font-size:11pt"><span style="line-height:107%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:Calibri,sans-serif"><b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">سابقه و هدف:</span></span></span></b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> باکتری</span></span></span> <i><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">اشریشیاکلی</span></span></span></i> <span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">O157:H7 </span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">یکی از خطرناک‌ترین باکتری‌های بیماری‌زا است که از طریق غذا در سراسر جهان منتقل می‌شود.</span></span></span> <span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">بسیاری از تکنیک‌های تشخیصی مبتنی بر تشخیص مولکولی و سرولوژیکی</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> محدودیت‌هایی دارند. در چند سال اخیر، آپتامرها به‌عنوان یک نوع اسیدنوکلئیک کوچک، در بسیاری از کاربردهای مختلف به‌کار گرفته شده‌اند و در سیستم‌های مختلف تشخیص، نقش مهمی به‌عنوان ابزار غربالگری داشته‌اند. در این مطالعه، با هدف مقایسۀ دو آپتامر و شناسایی سلول باکتری <i>اشریشیاکلی</i> انجام شده‌است.</span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:107%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:Calibri,sans-serif"><b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">مواد و روش‌ها</span></span></span></b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">: طراحی آغازگر اختصاصی برای آپتامرها با نرم‌افزار </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Oligo 7</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> انجام شد. حد تشخیص و غلظت بهینۀ آپتامر با روش </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">ELISA</span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> تعیین شد و میل ترکیبی آپتامر به سلول باکتری </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">&nbsp;O157:H7</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">با استفاده از دستگاه پلاسمای رزونانس سطحی بررسی شد.</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"B Nazanin""></span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:107%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:Calibri,sans-serif"><b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">یافته‌ها:</span></span></span></b> <span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">حد تشخیص برای آپتامر </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E. Coli 73</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> و </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E. Coli 81</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">،</span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">CFU/mL </span></span></span><span style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">&nbsp;10³به‌دست آمد و میل اتصال آپتامر </span></span></span>&nbsp;<span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E.<i> </i>Coli 73</span></span></span><i><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">و</span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E. coli 81</span></span></span></i><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> به </span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">سلول باکتری</span></span></span> <span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">به‌ترتیب </span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">8</span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E-</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> 1674/5 </span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">و </span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">9</span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E -</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> 8137/1</span></span></span> <span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">مولار</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> محاسبه شد. </span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:107%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:Calibri,sans-serif"><b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">نتیجه‌گیری</span></span></span></b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">: هر دو آپتامر میل اتصال خوبی را به سلول باکتری </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">O157:H7</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> نشان دادند. اما آپتامر </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E. Coli 81</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> دارای حساسیت و ویژگی بیشتری نسبت به آپتامر </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E. Coli 73</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> بود.</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> ازاین‌رو، آپتامر </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">E. Coli 81</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> می‌تواند به‌عنوان یک پیشنهاد مناسب برای تشخیص سلول‌های باکتری </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">O157:H7</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:"> در آینده مطرح شود.</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"B Nazanin""></span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:107%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:Calibri,sans-serif"><b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">واژگان کلیدی</span></span></span></b><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">: آپتامر، باکتری <i>اشریشیاکلی </i></span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">O157:H7</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">، </span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">ELISA</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span b="" nazanin="" style="font-family:">، پلاسمای رزونانس سطحی</span></span></span><span dir="LTR" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">.</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="line-height:107%"><span style="font-family:"B Nazanin""></span></span></span></span></span></span></span></span><br> &nbsp; <span style="font-size:11pt"><span style="unicode-bidi:embed"><span style="line-height:107%"><span style="font-family:Calibri,sans-serif"><b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.1pt">Aim and Background</span></span></span></span></b><i><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">:<span style="background:white"><span style="color:#1c1917"> Escherichia coli </span></span></span></span></span></i><span style="font-size:10.0pt"><span style="background:white"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:#1c1917">O157:H7 is one of the most dangerous foodborne pathogenic bacteria transmitted through food worldwide. Many detection techniques based on molecular and serological detection have limitations.</span></span></span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""> <span style="background:white"><span style="color:#1c1917">In recent years, aptamers as small nucleic acid molecules have been utilized in many different applications and played an essential role as screening tools in various diagnostic systems. In this study, two aptamers were compared and evaluated for the detection of &nbsp;<i>Escherichia coli</i> cells.</span></span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="unicode-bidi:embed"><span style="line-height:107%"><span style="font-family:Calibri,sans-serif"><b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.1pt">Material and Methods</span></span></span></span></b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.1pt">:</span></span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""> <span style="background:white"><span style="color:#1c1917">Specific primers were designed for each aptamer using the Oligo 7 software. The enzyme-linked immunosorbent assay (ELISA) was used to determine the detection limit and optimal concentration of the aptamers. The binding affinity of the aptamers to O157:H7 bacterial cells was investigated using a surface plasmon resonance instrument.</span></span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="unicode-bidi:embed"><span style="line-height:107%"><span style="font-family:Calibri,sans-serif"><b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.1pt">Results</span></span></span></span></b><b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">:</span></span></span></b><span style="font-size:10.0pt"><span style="background:white"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:#1c1917"> The detection limits for E<i>. </i>Coli 73 and E. Coli 81 aptamers were determined to be 10³ CFU/mL</span></span></span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">. </span></span></span><span style="font-size:10.0pt"><span style="background:white"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="color:#1c1917">The binding affinities of the E<i>. </i>Coli 73 and E. Coli 81 aptamers were 1.8137 E -9 and 5.1674E-8 mol/L, respectively.</span></span></span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.1pt"></span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="unicode-bidi:embed"><span style="line-height:107%"><span style="font-family:Calibri,sans-serif"><b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.1pt">Conclusion</span></span></span></span></b><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""><span style="letter-spacing:-.1pt">:</span></span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""> Both aptamers showed good binding affinity &nbsp;to O157:H7 bacterial cells. However, the E. Coli<i> </i>81 aptamer had higher sensitivity and specificity compared to the E. Coli<i> </i>73 aptamer. Therefore, the E. Coli 81 aptamer could be a suitable candidate for the detection of &nbsp;O157:H7 bacterial cells in future applications.</span></span></span></span></span></span></span><br> &nbsp; آپتامر, باکتری اشریشیاکلی O157:H7, ELISA, پلاسمای رزونانس سطحی. Aptamer, Escherichia coli O157:H7, ELISA, Surface resonance plasma 95 106 http://ncmbjpiau.ir/browse.php?a_code=A-10-1460-2&slc_lang=other&sid=1 Nafiseh Shafiei نفیسه شفعی 100319475328460014949 100319475328460014949 No Hamideh Mahmoodzadeh Hosseini حمیده محمود زاده حسینی hosseini361@yahoo.com 100319475328460014950 100319475328460014950 Yes Applied Microbiology Research Center, Biomedicine Technologies institute, Baqiyatallah University of Medical Sciences, Tehran, Iran مرکز تحقیقات میکروبیولوژی کاربردی، پژوهشکدۀ فناوری‌های زیست‌پزشکی، دانشگاه علوم پزشکی بقیه الله(عج)، تهران، ایران Jafar Amani جعفر امانی 100319475328460014951 100319475328460014951 No Seyed Ali Mirhosseini سید علی میرحسینی 100319475328460014952 100319475328460014952 No Hanieh Jafary هانیه جعفری 100319475328460014953 100319475328460014953 No