Effective Lux (Light Upon eXtension) Primer System for Early and Rapid Detection of<i>Coxiella burnetii</i>

Kunchev, Metodi; Alexandrov, E.; Kamarinchev, Bojin; Marinov, Krustyu; Gergova, Ivanka; Mekouchinov, Krassimir

doi:10.1080/13102818.2007.10817470

Cited by 2 publications

(1 citation statement)

References 17 publications

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“…LUX™ primers have been designed for use in a number of studies mainly in virology (Aitichou et al 2005;Antal et al 2007;Chen et al 2004;Nordgren et al 2008;Slavov et al 2008). However, some applications in bacteriology have been reported (Balcazar et al 2007;Kunchev et al 2007;McCrea et al 2007;Mitchell et al 2009;Xu et al 2008).…”

Section: Introductionmentioning

confidence: 96%

Comparison of In-house and Commercial Real-time PCR Systems for the Detection of Enterobacteriaceae and their Evaluation Within an Interlaboratory Study Using Infant Formula Samples

2011

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Traditional detection methods for Enterobacteriaceae in foods are time-consuming and laborious. The current study assessed the specificity of three real-time PCR primer sets. Set A (IEC primers) targeted the conserved flanking regions of the 16S rRNA, the 16S-ITS-23S gene region. Set B (ENT primers) annealed to Escherichia coli 16S ribosomal RNA gene. The third set (C) used a D-LUX™ (Light Upon eXtension) single FAMlabelled forward primer and a corresponding unlabeled primer. Set A was specific for E. coli and for some nonEnterobacteriaceae. SYBR Green-based real-time PCR confirmed the specificity of set B for the Enterobacteriaceae but also detected Vibrionaceae. In contrast, set C was poorly specific. However, set D including the forward LUX™ primer from set C and the reverse primer from set B had a specificity comparable to that of set B, but with higher sensitivity. This combined set was successfully applied to detect Enterobacteriaceae in infant milk formula and compared favourably with a commercial real-time PCR kit.

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