fa ارزیابی روش PCR جهت تشخیص آلودگی به باکتری سالمونلا انتریتیدیس در محصولات ماکیان در شهرستان کرج ميكروبيولوژي Microbiology بنيادي Basic <p align="justify"> </p><p class="a" dir="rtl" style="MARGIN: 0cm 0cm 0pt LINE-HEIGHT: normal" align="justify"><span lang="FA" style="FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-bidi-font-weight: bold mso-no-proof: yes"><strong>سابقه و هدف</strong></span><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt">:</span></i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-ansi-font-weight: bold"> </span><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt">باکتری سالمونلا انتریکا سرووار انتریتیدیس</span></i><span dir="ltr"></span><i><span lang="FA" dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'"><span dir="ltr"></span><font face="Times New Roman"> </font></span></i><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt">یکی از سویه­های شایع درگیر کننده</span></i><span dir="ltr"></span><i><span lang="FA" dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'"><span dir="ltr"></span><font face="Times New Roman"> </font></span></i><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt">ماکیان است که از طریق مواد غذایی آلوده قابل انتقال</span></i><span dir="ltr"></span><i><span lang="FA" dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'"><span dir="ltr"></span><font face="Times New Roman"> </font></span></i><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt">به انسان می­باشد. در این مطالعه از روش </span></i><font face="Times New Roman"><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'">PCR</span></i><span dir="rtl"></span></font><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span> برای تشخیص باکتری سالمونلا انتریتیدیس در محصولات ماکیان استفاده شد.</span></i><i><span dir="ltr" style="FONT-SIZE: 10pt LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 14.0pt mso-bidi-font-family: 'B Nazanin'"><p></p></span></i></p><p class="a" dir="rtl" style="MARGIN: 0cm 0cm 0pt LINE-HEIGHT: normal" align="justify"><span lang="FA" style="FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-bidi-font-weight: bold mso-no-proof: yes"><strong>روش بررسی</strong></span><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt">: در این پژوهش بنیادی، 80 نمونه از محصولات ماکیان از مراکز عرضه در شهرستان کرج و توابع آن تهیه گردید. </span></i><font face="Times New Roman"><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'">DNA</span></i><span dir="rtl"></span></font><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span> نمونه­ها</span></i><span dir="ltr"></span><i><span lang="FA" dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'"><span dir="ltr"></span><font face="Times New Roman"> </font></span></i><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt">به روش</span></i><font face="Times New Roman"><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'">salting out </span></i><span dir="rtl"></span></font><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span><font size="3"><font face="B Nazanin"><span style="mso-spacerun: yes"> </span>استخراج شد و </font></font></span></i><font face="Times New Roman"><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'">PCR</span></i><span dir="rtl"></span></font><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span> با استفاده از پرایمرهای اختصاصی مربوط به ژن سنتز تاژک به عنوان توالی هدف و سویه استاندارد سالمونلا انتریتیدیس (1621 </span></i><font face="Times New Roman"><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'">RTCC</span></i><span dir="rtl"></span></font><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span>) به عنوان کنترل مثبت بهینه­سازی و انجام گردید. <p></p></span></i></p><p class="a" dir="rtl" style="MARGIN: 0cm 0cm 0pt LINE-HEIGHT: normal" align="justify"><span lang="FA" style="FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-bidi-font-weight: bold mso-no-proof: yes"><strong>یافته</strong></span><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 14pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt mso-ansi-font-weight: bold">­</span><span lang="FA" style="FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-bidi-font-weight: bold mso-no-proof: yes"><strong>ها:</strong></span><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"> بررسی محصولات </span></i><font face="Times New Roman"><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'">PCR</span></i><span dir="rtl"></span></font><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span> به روش الکتروفورز نشان از تشکیل قطعه 250 جفت بازی و آلودگی به سالمونلا انتریتیدیس در 16 مورد از نمونه­های گوشت مرغ (40 درصد) و 9 مورد<font size="3"><font face="B Nazanin"><span style="mso-spacerun: yes">  </span>از نمونه­های تخم مرغ (23 درصد) داشت. در آزمون تعیین ویژگی، نتیجه تست در مورد 6 باکتری روده­ای دیگر از سایر سویه­ها و جنس­ها منفی گردید و</font></font></span></i><span dir="ltr"></span><i><span lang="FA" dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'"><span dir="ltr"></span><font face="Times New Roman"> </font></span></i><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt">حساسیت روش</span></i><font face="Times New Roman"><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'">PCR </span></i><span dir="rtl"></span></font><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span><font size="3"><font face="B Nazanin"><span style="mso-spacerun: yes"> </span>در این</font></font></span></i><span dir="ltr"></span><i><span lang="FA" dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'"><span dir="ltr"></span><font face="Times New Roman"> </font></span></i><span dir="rtl"></span><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span><font size="3"><font face="B Nazanin"><span style="mso-spacerun: yes"> </span>بررسی در سطح</font></font></span></i><span dir="ltr"></span><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'"><span dir="ltr"></span><font face="Times New Roman"> DNA<span style="mso-spacerun: yes"><font size="2">  </font></span></font></span></i><span dir="rtl"></span><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span><font size="3"><font face="B Nazanin"><span style="mso-spacerun: yes"> </span>ژنومی </font></font></span></i><font face="Times New Roman"><i><span dir="ltr" style="FONT-WEIGHT: normal FONT-SIZE: 10pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin'">fg</span></i><span dir="rtl"></span></font><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><span dir="rtl"></span> 100 تعیین شد.</span></i></p><p class="a" dir="rtl" style="MARGIN: 0cm 0cm 0pt LINE-HEIGHT: normal" align="justify"><i><span lang="FA" style="FONT-WEIGHT: normal FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt"><strong>نتیجه گیری</strong>:این مطالعه نشان داد که با توجه به محدودیت­ها و مشکلات روش­های جداسازی آزمایشگاهی می­توان از روش   <span style="FONT-SIZE: 10pt FONT-FAMILY: 'Times New Roman','serif' mso-bidi-language: FA mso-bidi-font-weight: bold mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin' mso-fareast-font-family: 'Times New Roman' mso-ansi-language: EN-US mso-fareast-language: EN-US">PCR</span></span></i></p><p align="justify"><i><span lang="FA" dir="rtl" style="FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt mso-ansi-font-weight: bold mso-fareast-font-family: 'Times New Roman' mso-ascii-font-family: 'Times New Roman' mso-hansi-font-family: 'Times New Roman' mso-ansi-language: EN-US mso-fareast-language: EN-US"> <span lang="FA" dir="rtl" style="FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt mso-ansi-font-weight: bold mso-fareast-font-family: 'Times New Roman' mso-ascii-font-family: 'Times New Roman' mso-hansi-font-family: 'Times New Roman' mso-ansi-language: EN-US mso-fareast-language: EN-US">استفاده کرد</span></span><span lang="FA" dir="rtl" style="FONT-SIZE: 12pt FONT-FAMILY: 'B Nazanin' mso-bidi-language: FA mso-no-proof: yes mso-ansi-font-size: 10.0pt mso-ansi-font-weight: bold mso-fareast-font-family: 'Times New Roman' mso-ascii-font-family: 'Times New Roman' mso-hansi-font-family: 'Times New Roman' mso-ansi-language: EN-US mso-fareast-language: EN-US"> به عنوان روشی حساس و سریع در تشخیص آلودگی نمونه­های غذایی به باکتری <span style="LETTER-SPACING: -0.2pt"><font face="B Nazanin" size="3">سالمونل انتریتیدیس</font></span></span><span dir="ltr"></span><b style="mso-bidi-font-weight: normal"><span lang="FA" style="FONT-SIZE: 10pt FONT-FAMILY: 'Times New Roman','serif' LETTER-SPACING: -0.2pt mso-bidi-language: FA mso-no-proof: yes mso-bidi-font-size: 12.0pt mso-bidi-font-family: 'B Nazanin' mso-fareast-font-family: 'Times New Roman' mso-ansi-language: EN-US mso-fareast-language: EN-US"><span dir="ltr"></span> </span></b></i></p> <p><strong>Background:</strong> Salmonella enterica serovar Enteritidis which involves poultry is transmitted by food. The aim of this survey was to optimize PCR method in order to detect Salmonella enteritidis in poultry products. </p><p><strong>Materials and methods</strong>: In this basic research, 80 samples (40 broiler meat and 40 egg samples) were obtained from distribution centers in Karaj and suburb. Following the DNA extraction of the samples, PCR was optimized and performed using standard strain of Salmonella enteritidis (RTCC1621) as positive control and primers against the flagella coding sequence of Salmonella Enteritidis genome.</p><p><strong> Results:</strong> The analysis of the PCR products by agarose gel electrophoresis indicated the amplification of 250 bp segments in 16 out of 40 (40%) broiler meat and 9 out of 40 (23%) egg samples. The sensitivity of the PCR at the DNA level was found to be 1pg and the specificity of the PCR was determined using 6 other entric Gram negative bacteria and found to be positive only for Salmonella enteritidis. </p><p><strong>Conclusion:</strong> This study confirmed that PCR provides sensitive, specific and rapid approach for detection of Salmonella enteritidis in food samples. </p> سالمونلا انتریتیدیس، PCR، محصولات ماکیان Salmonella enteritidis, PCR, poultry products 32 37 http://tmuj.iautmu.ac.ir/browse.php?a_code=A-10-1-336&slc_lang=fa&sid=1 Negar Nayebi نگار نایبی 10031947532846003085 10031947532846003085 Yes Islamic Azad University, Tehran, Iran گروه میکروبیولوژی، دانشگاه آزاد اسلامی، واحد کرج Seyed Ali Ghorashi سیدعلی قرشی 10031947532846003086 10031947532846003086 No National Institute of Genetic Engineering and Biotechnology, Tehran, Iran گروه زیست فناوری دام و آبزیان، پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری Naser Harzandi ناصر هرزندی 10031947532846003087 10031947532846003087 No Karaj Branch, Islamic Azad University, Tehran, Iran گروه میکروب شناسی، دانشگاه آزاد اسلامی، واحد کرج Mehdi Shamsara مهدی شمس آرا 10031947532846003088 10031947532846003088 No National Institute of Genetic Engineering and Biotechnology, Tehran, Iran گروه زیست فناوری دام و آبزیان، پژوهشگاه ملی مهندسی ژنتیک و زیست فناوری Bahman Tabarai بهمن تبرایی 10031947532846003089 10031947532846003089 No Vaccination Center, Pasteur Institute of Iran موسسه تحقیقاتی انستیتو پاستور ایران، بخش واکسن Amir Bakhtiari امیر بختیاری 10031947532846003090 10031947532846003090 No Islamic Azad University, Tehran, Iran دانشگاه آزاد اسلامی، واحد علوم و تحقیقات