کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
6132756 | 1593441 | 2016 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
HPV16 detection by qPCR method in relation to quantity and quality of DNA extracted from archival formalin fixed and paraffin embedded head and neck cancer tissues by three commercially available kits
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کلمات کلیدی
qPCRSCCFFPE tissueDNA qualityFFPE - MEPSpectrophotometer - اسپکتروفتومترbase pair - جفت پایهstandard error - خطای استانداردDouble stranded - دو رشتهformalin-fixed and paraffin-embedded - فرمالین ثابت و پارافین تعبیه شده استpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازHuman papillomavirus - ویروس پاپیلوم انسانیHPV - ویروس پایپلوم انسانیSquamous cell carcinoma - کارسینوم سلول سنگفرشی
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ویروس شناسی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The aim of the present study was to compare HPV16 detection by quantitative polymerase chain reaction (qPCR) in relation to the quantity and quality of DNA isolated from 21 formalin fixed and paraffin embedded (FFPE) head and neck cancer tissues by three commercially available kits: EX-WAX⢠DNA Extraction Kit (M) (Merck Millipore, Darmstadt, Germany), QIAamp® DNA FFPE Tissue (Q) (Qiagen, Hilden, Germany) and ReliaPrep⢠FFPE gDNA Miniprep System (P) (Promega, Madison, USA). Quantity of extracted DNA was assessed spectrophometrically and fluorometrically. Its quality was analyzed using A260/280 and A260/230 ratios and the β-actin fragment amplifiability in qPCR. HPV16 presence was detected by qPCR, using specific primers and TaqMan probe. HPV infection was found in 8 DNA samples extracted with M kit (38.1%) and in 7 (33.3%) isolated with Q and P kits. Three samples from M and Q kits were characterized by HPV16 positivity and lack of β-actin amplifiability. They had significantly lower A260/280 ratio (M: 1.6 ± 0.0, p = 0.044 and Q: 1.7 ± 0.0, p = 0.016) compared to samples with both fragments amplification (M: 1.7 ± 0.0 and Q: 1.9 ± 0.0). Therefore, for HPV detection by qPCR in FFPE tissues we recommend ReliaPrep⢠FFPE gDNA Miniprep System.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Virological Methods - Volume 236, October 2016, Pages 157-163
Journal: Journal of Virological Methods - Volume 236, October 2016, Pages 157-163
نویسندگان
Beata Ph.D., D.Sc., Anna Janecka, Anna Mucha-MaÅecka, Agnieszka Adamczyk, SÅawa Szostek, Dorota SÅonina, Krzysztof Halaszka, Marcin Przewoźnik,