Journal of Iranian Blood Transfusion
فصلنامه پژوهشی خون
bloodj
Medical Sciences
http://bloodjournal.ir
91
journal91
1027-9520
1735-8248
10.61882/bloodj
fa
jalali
1403
3
1
gregorian
2024
6
1
21
2
online
1
fulltext
fa
تشخیص مولکولی پروویروس HTLV-1 از DNA آزاد شده از سلولها در پلاسما، در موارد عدم دسترسی به DNA ژنومی سلولهای تک هستهای خون محیطی
Molecular detection of HTLV-1 provirus from DNA released from cells in plasma, in cases of lack of access to genomic DNA of peripheral blood mononuclear cells
ويروس شناسي
Virology
پژوهشي
Research
<div><span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><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="line-height:80%"><span style="font-family:Jadid"><span style="font-weight:normal">چکید</span></span></span><span lang="AR-SA" style="line-height:80%"><span style="font-family:Jadid"><span style="font-weight:normal">ه</span></span></span><span lang="FA" style="line-height:80%"><span style="font-family:Jadid"><span style="font-weight:normal"></span></span></span></span></span></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><i><span lang="FA" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:Titr">سابقه و هدف </span></span></span></i></b></span></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">ویروس لنفوتروپیک تیپ </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%">I</span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> انسانی، عمدتاً لنفوسیت­ها را آلوده می­کند. برای </span></span></span></b><b><i><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">تشخیص آین ویروس</span></span></span></i></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> ، عمدتاً سلولهای هسته دار مورد ارزیابی قرار میگیرد. با توجه به طول عمر کوتاه سلول ها، جمعآوری نمونه یک چالش به حساب می­آید. هدف از این مطالعه، بررسی حضور ویروس</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""> HTLV-1 </span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">در</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""> DNA </span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">ژنومی آزاد شده از سلول ها در پلاسما و تعیین بار ویروسی آن در اهداکنندگان خون وسترن بلات مثبت بود</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">.</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""></span></span></span></b></span></span></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><i><span lang="FA" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:Titr">مواد و روشها</span></span></span></i></b></span></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">در این مطالعه توصیفی، 30 اهداکننده ناقل ویروس</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""> HTLV-1 </span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">وسترن بلات مثبت بررسی شدند. با استفاده از کیت استخراج، </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">DNA</span></span></span></b><b><span style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> ژنومی از پلاسما، </span></span></span></b><b> </b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">و </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">PBMC</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> جداسازی شده با فایکول، استخراج گردید. حضور ویروس</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""> HTLV-1 </span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">با آزمایش مولکولی </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""> Nested PCR</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> تایید و جهت تعیین بار ویروس از آزمایش کمی </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">Real-Time Sybergreen PCR</span></span></span></b><b><span style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> استفاده گردید</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">.</span></span></span></b><b> </b></span></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><i><span lang="FA" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:Titr">یافتهها</span></span></span></i></b></span></span></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">میانگین سنی افراد برابر با 5/13 </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">±</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> 9/42 سال بود. 3/93% از افراد مذکر و 6/6% مؤنث بودند. ناحیه ژنی</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""> LTR </span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">به روش</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""> Nested PCR </span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">در80% از نمونه­های</span></span></span></b><b> </b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">پلاسما و 100% نمونه­های</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold""> PBMC </span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">شناسایی گردید. میانه لود ویروس در پلاسما برابر با </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">Copies/µL</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> 90/1 و میانه لود ویروس در </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">PBMC</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> برابر با </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">PBMC</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> 10<sup>6</sup>/ </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">Copies</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> 20 بود. </span></span></span></b></span></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><i><span lang="FA" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:Titr">نتیجه گیری</span></span></span></i></b></span></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:inter-ideograph"><span style="line-height:80%"><span style="direction:rtl"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><a name="_Hlk165009536"><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus"">جهت تشخیص مولکولی ویروس </span></span></span></b></a><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">HTLV-1</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> ، میتوان پروویروس </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">HTLV-1</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> را در </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">DNA</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> آزاد شده از سلولها در پلاسما، در مواردی که سلولهای تک هستهای خون محیطی در دسترس نمیباشد، مورد استفاده قرار داد. همچنین دیده شد که میزان بار ویروس در پلاسمای ناقلین بدون علامت نسبت به </span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">PBMC</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""> حدود 10 برابر کمتر می­باشد</span></span></span></b><b><span dir="LTR" style="font-size:9.0pt"><span style="line-height:80%"><span style="font-family:"Times New Roman Bold"">.</span></span></span></b><b><span lang="FA" style="font-size:11.0pt"><span style="line-height:80%"><span style="font-family:"2 Lotus""></span></span></span></b></span></span></span></span></span></span><br>
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</div>
<div dir="rtl" style="text-align: left;"><span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><span style="font-size:12.0pt">Abstract</span></b><b><span style="font-size:12.0pt"></span></b></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><i>Background and Objectives</i></b></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif"">Human lymphotropic virus type I mainly infects lymphocytes. For the detection of this virus, mainly nucleated cells are evaluated. Due to the short longevity of cells, sample collection is a challenge. The purpose of the study was to investigate the presence of HTLV-1 virus in genomic DNA released from cells in plasma and to determine its viral load in Western blot-positive blood donors<span dir="RTL" lang="AR-SA" style="font-family:"Traditional Arabic"">.</span></span></span></span></span></span><br>
<br>
<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><i>Materials and Methods</i></b></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif"">In this descriptive study, 30 HTLV-1 positive Western blot donors were evaluated. Using an extraction kit, genomic DNA was extracted from plasma and isolated PBMC using Ficol. A nested PCR test was used to confirm the presence of HTLV-1 virus. A real-time PCR quantitative test by Sybergreen method was used to determine the amount of virus load<span dir="RTL" lang="AR-SA" style="font-family:"Traditional Arabic"">.</span></span></span></span></span></span><br>
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<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><i>Results</i></b></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif"">The average age of subjects was 42.9 ± 13.5 years; 93.3% of them were male and 6.6% female. LTR gene region was detected by nested PCR method in 80% of plasma and 100% of PBMC samples. The median viral loads in plasma and PBMC were 1.90 Copies/µL and 20 copies/10<sup>6</sup> PBMC, respectively.</span></span></span></span></span><br>
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<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif""><b><i>Conclusions </i></b></span></span></span></span></span><br>
<span style="font-size:10pt"><span style="text-justify:kashida"><span style="text-kashida:0%"><span style="unicode-bidi:embed"><span style="font-family:"Times New Roman","serif"">To detect the HTLV-1 virus by molecular method, the HTLV-1 provirus DNA released from cells, in the plasma can be used when peripheral blood mononuclear cells are unavailable. It has also been observed that the virus load in the plasma of asymptomatic carriers is approximately 10 times lower than in PBMC.</span></span></span></span></span><br>
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کلمات کلیدی: ویروس لنفوتروپیک انسانی تیپ I , اسید نوکلئیک آزاد شده از سلول, PCR کمی
HTLV-1, Cell-Free Nucleic Acids, qPCR
138
150
http://bloodjournal.ir/browse.php?a_code=A-10-53-9&slc_lang=fa&sid=1
M.
Masoudi
منیره
مسعودی
9100319475328460024938
9100319475328460024938
No
مرکز تحقیقات انتقال خون ـ مؤسسه عالی آموزشی و پژوهشی طب انتقال خون
Z.
Sharifi
زهره
شریفی
sharifiz@yahoo.com
9100319475328460024939
9100319475328460024939
Yes
استاد مرکز تحقیقات انتقال خون ـ مؤسسه عالی آموزشی و پژوهشی طب انتقال خون
M.
Maghsudlu
مهتاب
مقصودلو
9100319475328460024940
9100319475328460024940
No
استاد مرکز تحقیقات انتقال خون ـ مؤسسه عالی آموزشی و پژوهشی طب انتقال خون