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Comparison of DNA extraction methods and gene targets from Leptospira interrogans serovar icterohaemorrhagiae for application in polymerase chain reaction

Indian Agricultural Research Journals

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Title Comparison of DNA extraction methods and gene targets from Leptospira interrogans serovar icterohaemorrhagiae for application in polymerase chain reaction
 
Creator ANITHA, S
RAJ, G DHINAKAR
RAJA, A
THANGAVELU, A
 
Subject Analytical sensitivity; DNA extraction methods; Gene targets; Leptospira; Polymerase chain reaction
 
Description The objective of this study was to identify the appropriate DNA extraction method and gene target from Leptospira interrogans serovar icterohaemorrhagiae for subsequent application in polymerase chain reaction (PCR) for diagnosis. DNA was extracted from pure cultures of Leptospira interrogans serovar icterohaemorrhagiae, spiked serum and urine samples using 4 different protocols,viz. STET buffer method, boiling method, nucleospin kit and the QIAamp DNA extraction kit using LipL41, LigA/B and LipL32 genes as targets. The lowest concentration of DNA that showed a visible band in PCR was taken as its analytical sensitivity in terms of concentration of DNA for each type of sample, target gene and DNA extraction method respectively. It was observed that DNA extraction from suspected clinical human sera samples using the QIAamp kit with either LipL41 or LipL32 as targets were most suitable for diagnosis by PCR since it had the greatest sensitivity of detection in PCR and the lowest amount of contaminating protein in the extracted DNA.
 
Publisher Directorate of Knowledge Management in Agriculture
 
Contributor —
 
Date 2012-10-11
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
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Format application/pdf
 
Identifier http://epubs.icar.org.in/ejournal/index.php/IJAnS/article/view/24278
 
Source The Indian Journal of Animal Sciences; Vol 82, No 10 (2012)
0367-8318
 
Language eng
 
Relation http://epubs.icar.org.in/ejournal/index.php/IJAnS/article/view/24278/11510
 
Rights Copyright (c) 2014 The Indian Journal of Animal Sciences