DETECTION OF DOUBLE-STRANDED-RNA BY ELISA AND DOT IMMUNOBINDING ASSAY USING AN ANTISERUM TO SYNTHETIC POLYNUCLEOTIDES

被引：12

作者：

ARAMBURU, J ^{[1
]}

NAVASCASTILLO, J ^{[1
]}

MORENO, P ^{[1
]}

CAMBRA, M ^{[1
]}

机构：

[1] INST VALENCIANO INVEST AGRARDAS,MONCADA,SPAIN

来源：

JOURNAL OF VIROLOGICAL METHODS | 1991年 / 33卷 / 1-2期

关键词：

DOUBLE-STRANDED RNA; DOT IMMUNOBINDING ASSAY; ELISA; SEROLOGICAL DETECTION;

D O I：

10.1016/0166-0934(91)90002-H

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An antiserum against polyinosinic-polycytidylic acid (In-Cn) was used to detect double-stranded RNA (dsRNA) by several serological techniques. DsRNA was readily detected by indirect ELISA (ELISA-I) and dot immunobinding assay (DIA). Addition of the antigen to poly-L-lysine-precoated plates and blocking with uncreamed milk powder allowed detection levels of 100 pg.ml-1 In-Cn by ELISA-I. Concentrations as low as 1 ng.ml-1 were detected by DIA using polyvinyliden difluoride (PVDF) membranes. Detection capacity with nitrocellulose membranes was 1000 times lower than with PVDF. ELISA-I and DIA enabled detection of dsRNA in enriched fractions from cucumber mosaic virus (CMV)- and citrus tristeza virus (CTV)-infected plants and from virus-infected Penicillium chrysogenum mycelium. These techniques showed similar or higher sensitivity for detection of dsRNA than separation by polyacrylamide gel electrophoresis and silver staining.

引用

页码：1 / 11

页数：11

共 19 条

[1] USE OF MONOCLONAL-ANTIBODY AGAINST POLY [I] - POLY [C] FOR DETECTING MYCOVIRUSES AND POTENTIAL APPLICATIONS TO POTATO SPINDLE TUBER VIROID AND ANIMAL REOVIRUSES [J].