کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
3453804 1595938 2016 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A novel porcine cell culture based protocol for the propagation of hepatitis E virus
موضوعات مرتبط
علوم پزشکی و سلامت پزشکی و دندانپزشکی پزشکی و دندانپزشکی (عمومی)
پیش نمایش صفحه اول مقاله
A novel porcine cell culture based protocol for the propagation of hepatitis E virus
چکیده انگلیسی

ObjectiveTo present a comprehensive protocol for the processing of hepatitis E virus (HEV) infected samples and propagation of the virus in primary cell cultures.MethodsHepatitis E was extracted from porcine liver and faecal samples following standard protocols. The virus was then allowed to attach in the presence of trypsin to primary cells that included porcine and bovine intestinal epithelial cells and macrophages over a period of up to 3 h. The virus was propagated by rotational passaging through the cell cultures. Propagation was confirmed by immunoblotting.ResultsWe developed a comprehensive protocol to propagate HEV in porcine cell model that includes (i) rotational culturing of the virus between porcine cell types, (ii) pre-incubation of infected cells for 210 min, (iii) use of a semi-complete cell culture medium supplemented with trypsin (0.33 µg/mL) and (iv) the use of simple immunoblot technique to detect the amplified virus based on the open reading frame 2/3.ConclusionsThis protocol opens doors towards systematic analysis of the mechanisms that underlie the pathogenesis of HEV in vitro. Using our protocol, one can complete the propagation process within 6 to 9 d.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Asian Pacific Journal of Tropical Disease - Volume 6, Issue 8, August 2016, Pages 596–602
نویسندگان
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