کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2089126 1545776 2007 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Immunochemical assay for deoxyribonuclease activity in body fluids
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی بیوتکنولوژی یا زیست‌فناوری
پیش نمایش صفحه اول مقاله
Immunochemical assay for deoxyribonuclease activity in body fluids
چکیده انگلیسی

We have developed two microtiter plate assays to quantify the deoxyribonuclease activity in biological fluids. Both assays are based on hydrolysis of biotinylated and fluorescein-labeled DNA substrates, with subsequent immunochemical detection of non-digested DNA. The assay based on hydrolysis of 974 bp PCR product labeled with biotinylated forward and fluorescein-labeled reverse primers is more sensitive (0.05 U/ml) and convenient for quantifying the DNase activity in biological fluids than the assay based on hydrolysis of double-labeled 20 bp oligonucleotide. The DNase activity in urine and blood plasma of healthy donors was measured using the PCR product-based assay. Urine samples revealed greater activity, 1.49 ± 1.41 U/ml; blood plasma DNase I-like activity was 0.36 ± 0.20 U/ml. DNase II-like activity was not detected in the plasma samples. The data obtained confirm that DNase I-like enzymes are responsible for the majority of deoxyribonuclease activity in blood plasma.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Immunological Methods - Volume 325, Issues 1–2, 31 August 2007, Pages 96–103
نویسندگان
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