Novel Light-Upon-Extension (LUX) Real-Time PCR Primer System for Rapid Detection of<i>Polyomavirus Hominis</i>1 (BKV) in Clinical Samples

Slavov, Svetoslav Nanev; Kalvatchev, Z.; Tsekov, Iliya; Simeonov, P; Hristova, L; Kotsev, J.; Mladenov, D; Tsvetkov, Mitko

doi:10.1080/13102818.2008.10817547

Cited by 3 publications

(3 citation statements)

References 6 publications

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“…Such fluorogenic PCR has the potential to be routinely used in food industries because of rapidity, simplicity, and lower cost compared with real-time PCR systems using probes for instance. 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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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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“…The melting curve post PcR analysis was performed in a similar manner as described above for BKV (8). the melting curve temperature of the JCV PCR product was ~78.4° C. Lack of specific increase in the fluorescent signal and no distinguishable melting peaks were observed in the CSF samples from people with other neurological disorders or with polyomavirus BK, SV-40, human papillomavirus 16 and 18, human herpes virus 1 DnA and in a template free control (Fig.…”

Section: Primer Design and Characteristics Of The Lux-primers Formentioning

confidence: 99%

“…the system has been applied for the specific amplification of several oncogenic human viruses including human papillomavirus type 16 (3) and the human polyoma virus BK (8). As far as our knowledge lUX technology based systems for other polyoma viruses have not been described in the literature.…”

Section: Introductionmentioning

confidence: 99%

Design and Evaluation of New Light-Upon-Extension (Lux) Real-Time PCR Primer System for Detection ofPolyomavirus Hominis 2(JC Virus) in Clinical Samples

Ferdinandov

Kalvatchev

Ivan

2009

Biotechnology & Biotechnological Equipment

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Do human polyoma viruses and human immunodeficiency virus share common co‐receptors?

Borissov

Tsekov

Gavazova

et al. 2009

Journal of Medical Virology

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Host and/or viral factors involved in human polyomavirus (HPoV) infection in persons living with HIV remain unknown. A hypothesis is outlined suggesting the importance of the co‐receptors CCR5, CCR2, and CXCR4 not only for HIV, but also for HPoV. Functionally capable receptors coded by wild‐type (wt) genotypes could facilitate internalization of HPoV in the cell resulting in brain and/or kidney infection/s in HIV infected individuals. Forty‐nine Bulgarians with HIV, all treated by HAART, without neurological and/or kidney disorders, were tested for JCV and BKV and genotyped for CCR5 (CCR5del32), CCR2 (CCR2‐64I), and CXCR4 (SDF1‐3′A). In 27/49 (55.1%) individuals a co‐infection with HPoV was identified—BKV in 12/49 (24.5%), JCV—in another 12/49 (24.5%), and both viruses—in 3/49 (6.1%). A high frequency of wt CCR5 was found in patients with HPoV (91.7% for BKV and JCV and in 100% with both viruses). V/V of CCR2 was presented in 75% for BKV and JCV and in 66.7% for BKV plus JCV. SDF1‐3′G/G predominated in JCV infected patients (75%), while G/A and A/A genotypes were more frequent in patients with BKV (41.7%). Also, 21/22 (95.4%) persons without HPoV infection were heterozygous for SDF1 and CCR2. The number of individuals bearing wt of all co‐receptors in the group of persons not infected with HPoV was lower (P = 0.03) than that with polymorphism/s in one or two genes (SDF1 and CCR2) in the same group. The results suggest a probable role of co‐receptors used by HIV to facilitate infection with HPoV. J. Med. Virol. 82:8–13, 2010. © 2009 Wiley‐Liss, Inc.

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Novel Light-Upon-Extension (LUX) Real-Time PCR Primer System for Rapid Detection ofPolyomavirus Hominis1 (BKV) in Clinical Samples

Cited by 3 publications

References 6 publications

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

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

Design and Evaluation of New Light-Upon-Extension (Lux) Real-Time PCR Primer System for Detection ofPolyomavirus Hominis 2(JC Virus) in Clinical Samples

Do human polyoma viruses and human immunodeficiency virus share common co‐receptors?

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