Production of positive controls for calcivirus-specific PCR using recombinant baculiovirus technology

Butcher, S.A. and Gould, E.A. (1997) Production of positive controls for calcivirus-specific PCR using recombinant baculiovirus technology. In: Sutcliffe, D.W. (ed.) The microbiological quality of water. Ambleside, UK, Freshwater Biological Association, pp. 48-53. (FBA Special Publications,5)

[img]
Preview
PDF (The document's language is English .) - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
Download (284Kb) | Preview

Abstract

Recent advances in our knowledge of the genetic structure of human caliciviruses (HuCVs) and small round-structured viruses (SRSVs) have led to the development of polymerase chain reaction (PCR)-based molecular tests specific for these viruses. These methods have been developed to detect a number of human pathogenic viruses in environmental samples including water, sewage and shellfish. HuCVs and SRSVs are not culturable, and no animal model is currently available. Therefore there is no convenient method of preparing viruses for study or for reagent production. One problem facing those attempting to use PCR-based methods for the detection of HuCVs and SRSVs is the lack of a suitable positive control substrate. This is particularly important when screening complex samples in which the levels of inhibitors present may significantly interfere with amplificiation. Regions within the RNA polymerase regions of two genetically distinct human caliciviruses have been amplified and used to produce recombinant baculoviruses which express RNA corresponding to the calicivirus polymerase. This RNA is being investigated as a positive control substrate for PCR testing, using current diagnostic primer sets. Recombinant baculovirus technology will enable efficient and cost-effective production of large quantities of positive control RNA with a specific known genotype. We consider the development of these systems as essential for successful screening and monitoring applications.

Item Type: Book Section
Title: Production of positive controls for calcivirus-specific PCR using recombinant baculiovirus technology
Personal Creator/Author:
CreatorsEmail
Butcher, S.A.
Gould, E.A.
Title of Book: The microbiological quality of water
Editors:
EditorsEmail
Sutcliffe, D.W.
Refereed: No
Series Name: FBA Special Publications
Number: 5
Page Range: pp. 48-53
Date: 1997
Publisher: Freshwater Biological Association
Place of Publication: Ambleside, UK
ISBN: 0-900386-57-6
Issuing Agency: Freshwater Biological Association
Uncontrolled Keywords: Polymerase chain reaction; Viruses; Viral diseases; RNA; Primers; Cloning; Diarrhetic shellfish poisoning; Public health; Sewages
Subjects: Health
Limnology
Pollution
Item ID: 5321
Depositing User: Hardy B Schwamm
Date Deposited: 11 May 2011 12:04
Last Modified: 29 Sep 2011 15:06
Related URLs:
URI: http://aquaticcommons.org/id/eprint/5321

Actions (login required)

View Item View Item

Document Downloads

More statistics for this item...