Use of touch-down polymerase chain reaction to enhance the sensitivity of Brucella melitensis detection in raw milk.

Animal Biotechnology
Heidar RahimiEhsan Rashidian

Abstract

Brucellosis is a highly contagious bacterial zoonotic infectious disease severely affecting the public health and economic features of endemic and non-endemic countries. The present study assessed the potentials of using the touch-down polymerase chain reaction (TD-PCR) compared to the conventional PCR and culture methods in order to detect Brucella melitensis in raw milk samples of 55 sheep and 45 goats through deriving the primers from the omp31 element of the Brucella genome. In addition, nine isolates of B. melitensis were identified using the culture method. No positive cases were found in sediment samples, while the fatty tap layer test by conventional PCR and TD-PCR revealed 6 and 16 positive samples, respectively. Based on the survey of the limits of detection by TD-PCR and conventional PCR, TD protocol had a detection threshold of three logs higher than the conventional protocol under the experimental condition. The developed protocol of this study was highly sensitive and extremely fast. Therefore, this TD-PCR protocol could detect even a very low number of bacteria in milk samples. To our best knowledge, this is the first report on the use of the TD-PCR method to identify B. melitensis in milk.

References

May 1, 1997·The Journal of Dairy Research·M AmillsA Sanchez
Feb 15, 2001·Current Opinion in Microbiology·M L BoschiroliD O'Callaghan
Jul 2, 2002·The Veterinary Journal·Mahmoud E R Hamdy, A S Amin
Nov 5, 2002·Veterinary Microbiology·Betsy J Bricker
Dec 5, 2002·The Journal of Dairy Research·Michael A MurphyChris Moran
Jun 11, 2005·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Martín J ZumárragaAngel Cataldi
Mar 21, 2006·Research in Veterinary Science·Seónadh O'LearyTorres Sweeney
May 17, 2011·Preventive Veterinary Medicine·J GodfroidJ-J Letesson
Sep 17, 2011·Vector Borne and Zoonotic Diseases·Ari A Shemesh, Pablo Yagupsky
Jul 11, 2013·Revue Scientifique Et Technique·S Al DahoukH Neubauer
May 21, 2014·Genetics and Molecular Research : GMR·T UsmanY Wang
Sep 23, 2014·International Journal of Systematic and Evolutionary Microbiology·Adrian M WhatmoreNatalia E Schlabritz-Loutsevitch
Feb 26, 2016·Animal Biotechnology·Joanna PokorskaJoanna Makulska
Mar 2, 2016·International Journal of Systematic and Evolutionary Microbiology·Holger C ScholzErwin Hofer
Mar 16, 2017·PLoS Neglected Tropical Diseases·Elisabeth Lindahl-RajalaUlf Magnusson
Sep 3, 2017·Journal of Microbiological Methods·Maria Cryskely A BatingaLara B Keid

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