Reverse transcription polymerase chain reaction method for the detection of glycopeptide resistance in enterococci

Privitera, O.; Sisto, Francesca; Giuffrida, Vincenza; Puntorieri, Miriam; Cascone, Carmela; Silvestro, Isabella Di; Rappazzo, Giancarlo; Stefani, Stefania

doi:10.1016/s0167-7012(98)00096-7

Cited by 4 publications

(1 citation statement)

References 9 publications

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“…Conventional RT‐PCR has been used to detect bacterial pathogens in food [13,19,20], to study glycopeptide resistance in enterococci [21] and enhanced expression of the multidrug efflux pump AcrAB in E. coli mutants selected with a fluoroquinolone [16] or the mexAB ‐ oprM multidrug efflux operon of Pseudomonas aeruginosa [22]. mRNA quantification can also be used to evaluate the expression of most virulence factors [12,23].…”

Section: Resultsmentioning

confidence: 99%

Comparison of two RT-PCR methods for quantifyingampCspecific transcripts inEscherichia colistrains

Corvec¹,

Caroff²,

Espaze³

et al. 2003

FEMS Microbiology Letters

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In Escherichia coli, beta-lactam resistance usually depends on beta-lactamase production. AmpC chromosomal cephalosporinase hyperproduction is generally due to mutations in the ampC gene promoter. In order to study ampC expression in E. coli clinical strains, we have compared two methods: conventional and real-time reverse transcription-polymerase chain reaction (RT-PCR). With both methods, ampC mRNA was found to be greatly increased in strains presenting -42 or -32 mutations in the ampC promoter, and moderately increased when a -11 mutation was present in the Pribnow box. Real-time RT-PCR represents a powerful tool combining amplification, fluorescent detection and analysis.

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Section: Resultsmentioning

confidence: 99%

Comparison of two RT-PCR methods for quantifyingampCspecific transcripts inEscherichia colistrains

Corvec¹,

Caroff²,

Espaze³

et al. 2003

FEMS Microbiology Letters

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Gene cloning and characterization of an aldehyde dehydrogenase from long-chain alkane-degrading Geobacillus thermoleovorans B23

et al. 2009

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Geobacillus thermoleovorans B23 is capable of degrading long-chain alkanes at 70 degrees C. Bt-aldh, an aldehyde dehydrogenase gene in B23, was located in the upstream region of p21 whose expression level was dramatically increased when alkane degradation was started (Kato et al. 2009, BMC Microbiol 9:60). Like p21, transcription level of Bt-aldh was also increased upon alkane degradation. Bt-Aldh (497 aa, MW = 53,886) was overproduced in Escherichia coli, purified, and characterized biochemically. Bt-Aldh acted as an octamer, required NAD(+) as a coenzyme, and showed high activity against aliphatic long-chain aldehydes such as tetradecanal. The optimum condition for activity was 50-55 degrees C and pH 10.0. The activity was elevated to two- to threefold in the presence of 2 mM Ba(2+), Ca(2+), or Sr(2+), while Mg(2+) and Zn(2+) inhibited the enzyme activity. Bt-Aldh represents thermophilic aldehyde dehydrogenases responsible for degradation of long-chain alkanes.

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Methods used for the isolation, enumeration, characterisation and identification of Enterococcus spp.

Domig

Mayer

Kneifel

2003

International Journal of Food Microbiology

103

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Reverse transcription polymerase chain reaction method for the detection of glycopeptide resistance in enterococci

Cited by 4 publications

References 9 publications

Comparison of two RT-PCR methods for quantifyingampCspecific transcripts inEscherichia colistrains

Comparison of two RT-PCR methods for quantifyingampCspecific transcripts inEscherichia colistrains

Gene cloning and characterization of an aldehyde dehydrogenase from long-chain alkane-degrading Geobacillus thermoleovorans B23

Methods used for the isolation, enumeration, characterisation and identification of Enterococcus spp.

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