Medical Sciences Journal of Islamic Azad University
فصلنامه علوم پزشکی دانشگاه آزاد اسلامی تهران
MEDICAL SCIENCES
Medical Sciences
http://tmuj.iautmu.ac.ir
1
admin
1023-5922
2008-3386
000
10.66224/iau
000
1023-5922
000
fa
jalali
1402
9
1
gregorian
2023
12
1
33
4
online
1
fulltext
fa
مقایسه روش سنجش ATP بیولومینسانس و روش استاندارد شمارش کلنی
جهت ارزیابی محصولات ب ث ژ
Comparison of the ATP-bioluminescence assay and the standard colony counting method for evaluation BCG products
داروسازي
Pharmacology
كيفي
Qualitative
<span style="font-size:11pt"><span style="line-height:normal"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman",serif"><span style="font-weight:bold"><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin"">سابقه و هدف</span></span><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:Nazanin"><span style="font-weight:normal">:</span></span></span></i> <i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">روش مرسوم سنجش کارایی محصولات واکسن ب ث ژ از طریق شمارش کلونی است که بسیار کند و طاقت فرسا است. به منظور کنترل کیفی واکسن، روشی سریع جهت تخمین باسیل های زنده مورد نیاز است. </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">ATP</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> شناساگر قابل اعتمادی برای قابلیت زنده مانی سلول است و میزان نور ساطع شده در واکنش بیولومینسانس بطور مستقیم وابسته به میزان </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">ATP </span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> باسیل ها است. در مطالعه حاضر رابطه میان دو روش فوق مورد بررسی قرار گرفت. </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal"></span></span></i></span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:normal"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman",serif"><span style="font-weight:bold"><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="letter-spacing:-.3pt">روش بررسی</span></span></span><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="letter-spacing:-.3pt"><span style="font-weight:normal">:</span></span></span></span></i> <i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">تعداد کلونی­های زنده (</span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">CFU</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">) و سیگنال بیولومینسانس (</span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">RLU</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">) محصولات </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">BCG</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> به ترتیب با استفاده از روش مرسوم میکروبیولوژیک و کیت اختصاصی تعیین شد. میزان </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">ATP</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> محصولات بر اساس میزان </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">ATP </span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> و </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">CFU</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> واکسن استاندارد </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">NIBSC</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> تخمین زده شد. آنالیز رگرسیون خطی جهت تعیین میزان همبستگی دو روش با استفاده از نرم افزار </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">Graph Pad Prism 8.0</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> انجام پذیرفت. ضریب </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">K </span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> جهت محاسبه تعداد کلونی محصولات </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">BCG </span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> تعیین شد. </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal"></span></span></i></span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:normal"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman",serif"><span style="font-weight:bold"><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="letter-spacing:-.2pt">یافته</span></span></span><span lang="FA" style="font-size:14.0pt"><span style="font-family:"B Nazanin""><span style="letter-spacing:-.2pt"><span style="font-weight:normal">­</span></span></span></span><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="letter-spacing:-.2pt">ها</span></span></span><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">:</span></span></span></i> <i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">ارتباط خطی و همبستگی قابل توجهی میان دو روش برای بالک ( 9874/0 =</span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">r<sup>2</sup></span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">) و واکسن </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">BCG </span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> ( 9417/0 =</span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">r<sup>2</sup></span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">) در مقایسه با پاستوسیس ( 7692/0 =</span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">r<sup>2</sup></span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">) بدست آمد. ضریب </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">K</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> برای واکسن و بالک </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">BCG</span></span></i><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> تعیین شد که می­توان از آن جهت پیش بینی و تخمین تعداد کلونی سری ساخت های جدید محصولات استفاده کرد. </span></span></span></i></span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:normal"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman",serif"><span style="font-weight:bold"><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin"">نتیجه</span></span><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal">­</span></span></span><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin"">گیری:</span></span><i><span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> این مطالعه اشاره می­کند که روش بهبود یافته بیولومینسانس، روشی سریع، حساس و قابل اعتماد جهت تسریع فرایند کنترل کیفی واکسن </span></span></span></i><i><span dir="LTR" style="font-size:10.0pt"><span style="font-weight:normal">BCG</span></span></i><i><span style="font-size:12.0pt"><span style="font-family:"B Nazanin""><span style="font-weight:normal"> است.</span></span></span></i></span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:11.0pt"><span style="font-family:"Times New Roman",serif"><b><span style="font-size:11.0pt">Background</span></b><span style="font-size:11.0pt">: The conventional method of evaluating the efficacy of BCG vaccine products is through colony count, which is very slow and cumbersome. A rapid test for estimating viable bacilli is necessary for vaccine quality control. ATP is a reliable marker for cell viability, and the amount of emitted light during bioluminescence reactions is directly proportional to the ATP content of the bacilli. In this study, the correlation between these two methods was evaluated. </span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:11.0pt"><span style="font-family:"Times New Roman",serif"><b><span style="font-size:11.0pt">Materials and methods</span></b><span style="font-size:11.0pt">: CFU (colony-forming unit) value and bioluminescence signal (RLU) of BCG products were determined by microbiological conventional method and the special kit respectively. ATP content of the products were estimated based on ATP content and CFU value of standard NIBSC vaccine. Linear regression analysis were done by Graph Pad Prism 8.0 software to determine the extent of correlation between two methods. The coefficient K was determined to calculate the CFU value of BCG products. </span></span></span></span></span></span><br>
<span style="font-size:12pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:11.0pt"><span style="font-family:"Times New Roman",serif"><b><span style="font-size:11.0pt">Results</span></b><span style="font-size:11.0pt"> Linear relationship and significant correlation were obtained between two methods for BCG Bulk (r<sup>2</sup>= 0.9874) and Vaccine (r<sup>2</sup>= 0.9417) compared to Pastocys (r<sup>2</sup>= 0.7692). The coefficients K were determined for BCG and Bulk vaccine (0.447 and 0.26 respectively) and can be used to predict and estimate the CFU value of new bathes of products. </span></span></span></span></span></span><br>
<b><span style="font-size:11.0pt"><span style="font-family:"Times New Roman",serif">Conclusion:</span></span></b><span style="font-size:11.0pt"><span style="font-family:"Times New Roman",serif"> The study suggests that an optimized ATP bioluminescence assay is a rapid, sensitive, and reliable alternative method that can accelerate the quality control process for BCG vaccine</span></span>
واکسن ب ث ژ, واحد تشکیل کلونی, سنجش ATP بیولومینسانس
BCG vaccine, Colony forming units, Bioluminescence ATP assay
365
371
http://tmuj.iautmu.ac.ir/browse.php?a_code=A-10-1619-1&slc_lang=fa&sid=1
Hamidreza
Hozouri
حمیدرضا
حضوری
h_hozouri@pasteur.ac.ir
100319475328460011806
100319475328460011806
Yes
Pasteur Institute of Iran, Production and Research Complex
استادیار، مجتمع تولیدی تحقیقاتی انستیتو پاستور ایران
Delaram
Doroud
دلارام
درود
d_doroud@pasteur.ac.ir
100319475328460011807
100319475328460011807
No
Pasteur Institute of Iran, Production and Research Complex
دانشیار، مجتمع تولیدی تحقیقاتی انستیتو پاستور ایران
Mohammadhossein
Hedayati
محمدحسین
هدایتی
mhheda@gmail.com
100319475328460011808
100319475328460011808
No
Expert in Cell Biology Laboratory, Pasteur Institute of Iran, Production and Research Complex
کارشناس آزمایشگاه بیولوژی سلولی مجتمع تولیدی تحقیقاتی انستیتو پاستور ایران