LCR Paper Abstract

Kevin O'Donnell odonnell at sasa.gov.uk
Wed Apr 17 04:07:35 EST 1996


Some time ago, I suggested that people could post abstracts of 
diagnostics papers here, in order to stimulate more on-topic 
discussion.  Here is an abstract from my own laboratory, of a paper 
publ;ished in the Proceedings of the Diagnostics in Crop Protection 
conference held in Warwick, UK earlier this month.

DETECTION OF POTATO VIRUS Y USING THE LIGASE CHAIN REACTION
 (LCR), IN COMBINATION WITH A MICROTITRE PLATE BASED METHOD
 FOR PRODUCT DETECTION 

K J O'DONNELL,  E CANNING  and L G A YOUNG 
Scottish Agricultural Science Agency, East Craigs, Edinburgh, EH12 8NJ, UK
email: odonnell at sasa.gov.uk
 

ABSTRACT
               
The Scottish Agricultural Science Agency (SASA) carries out diagnostic
 tests for viruses on potato leaf and tuber samples in support of the Scottish
 Seed Potato Classification Scheme. Currently, routine detection of potato
 viruses in submitted leaf samples is carried out using ELISA (enzyme-linked
 immunosorbent assay).  However, ELISA is not sufficiently sensitive 
to consistently detect the small amounts of virus present in primary infected
 tubers.  In this case, the dormancy of the tubers must first be broken and then 
the resulting plants tested by ELISA, a process which can take several weeks.

In order to reduce this period we have evaluated nucleic acid amplification
 techniques which can theoretically achieve the level of sensitivity necessary
 for virus detection directly from tubers. In this paper we report on the 
development of an assay for PVY based on the ligase chain reaction (LCR), 
a method based on the ligase-mediated exponential amplification of DNA probes
 specific to target DNA (or cDNA) of the pathogen. We have combined the
 LCR assay with a microtitre plate based detection system which removes the 
need to run electrophoresis gels to detect products of the assay.  This method
 has the possibility of combining the sensitivity and specificity of nucleic
 acid-based techniques with the automation of ELISA. Preliminary results are
 shown which compare the performance of LCR with the standard ELISA method. 

Dr Kevin O'Donnell                          "Nature is not cruel, only
Diagnostics and Molecular Biology    pitilessly indifferent."
SASA                                                   - Richard Dawkins
Edinburgh



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