scholarly journals Quinolone Resistance in Neisseria gonorrhoeae: Rapid Genotyping of Quinolone Resistance-Determining Regions in gyrA and parC Genes by Melting Curve Analysis Predicts Susceptibility

2009 ◽  
Vol 53 (3) ◽  
pp. 1264-1267 ◽  
Author(s):  
Frédérique Vernel-Pauillac ◽  
Tiffany R. Hogan ◽  
John W. Tapsall ◽  
Cyrille Goarant
Keyword(s):  
Public Health ◽  
Antibiotic Resistance ◽  
Neisseria Gonorrhoeae ◽  
Melting Curve ◽  
Curve Analysis ◽  
Quinolone Resistance ◽  
Fluoroquinolone Resistance ◽  
Melting Curve Analysis ◽  
Pcr Assay

ABSTRACT We report a duplex real-time PCR assay for the simultaneous screening of mutations involved in fluoroquinolone resistance within gyrA and parC quninolone resistance-determining regions (QRDRs) in Neisseria gonorrhoeae. Our assay clearly detects all mutated QRDRs and allows the identification of common genotypes, whether the QRDRs contain single or double mutations, providing valuable epidemiological tools. When this method is used in conjunction with similar assays and in vitro analyses, essential antibiotic resistance surveillance can be performed for public health purposes.

scholarly journals Rapid diagnosis of human brucellosis by SYBR Green I-based real-time PCR assay and melting curve analysis in serum samples

2005 ◽  
Vol 11 (9) ◽  
pp. 713-718 ◽  
Author(s):  
M.I. Queipo-Ortuño ◽  
J.D. Colmenero ◽  
J.M. Reguera ◽  
M.A. García-Ordoñez ◽  
M.E. Pachón ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Melting Curve ◽  
Rapid Diagnosis ◽  
Curve Analysis ◽  
Sybr Green I ◽  
Melting Curve Analysis ◽  
Human Brucellosis ◽  
Pcr Assay ◽  
Serum Samples

scholarly journals Real-Time Multiplex PCR Assay and Melting Curve Analysis for Identifying Diarrheagenic Escherichia coli

2013 ◽  
Vol 51 (3) ◽  
pp. 1031-1033 ◽  
Author(s):  
Tamara B. Souza ◽  
Diego M. Lozer ◽  
Sônia M. S. Kitagawa ◽  
Liliana C. Spano ◽  
Neusa P. Silva ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multiplex Pcr ◽  
Real Time ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Pcr Assay ◽  
Diarrheagenic Escherichia Coli ◽  
Multiplex Pcr Assay

scholarly journals Rapid detection and grouping of porcine bocaviruses by an EvaGreen ® based multiplex real-time PCR assay using melting curve analysis

2016 ◽  
Vol 30 (4) ◽  
pp. 195-204 ◽  
Author(s):  
Xiaowen Zheng ◽  
Gaopeng Liu ◽  
Tanja Opriessnig ◽  
Zining Wang ◽  
Zongqi Yang ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Pcr Assay

scholarly journals In Vitro Assessment of the Further Potential for Development of Fluoroquinolone Resistance in Neisseria meningitidis

2005 ◽  
Vol 49 (5) ◽  
pp. 1753-1760 ◽  
Author(s):  
Tiffany R. Shultz ◽  
Peter A. White ◽  
John W. Tapsall
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Neisseria Meningitidis ◽  
Quinolone Resistance ◽  
Clinical Isolates ◽  
Fluoroquinolone Resistance ◽  
In Vitro Assessment ◽  
Resistant Mutants ◽  
The Individual

ABSTRACT We examined the potential for the development of fluoroquinolone resistance in Neisseria meningitidis by cultivating two clinical isolates of meningococci in the presence of concentrations of ciprofloxacin at and about the predetermined MIC. The quinolone resistance determining regions (QRDRs) of gyrA and parC of 50 stable quinolone-resistant mutants derived in vitro were sequenced and compared with QRDR alterations reported in clinical isolates of quinolone-resistant meningococci and gonococci. MICs to ciprofloxacin and trovafloxacin were determined and sequence changes were correlated with quinolone MICs. Ciprofloxacin and trovafloxacin MICs of the in vitro-derived quinolone-resistant mutants ranged up to 16 mg/liter. Single GyrA alterations were the first change detected and were accompanied by raised MICs, followed by double GyrA changes and still higher MICs. MICs increased further as single ParC substitutions appeared and these were always in the presence of a single or double GyrA change. GyrA changes occurred at positions 91 and 95 with substitutions of Asp-95→Asn and Thr-91→Ala and Ile. Changes in the parC QRDR occurred at positions 85, 86, and 91 with four substitutions, Gly-85→Asp, Asp-86→Asn, Glu-91→Gly, and Glu-91→Lys, detected. The nature of the individual QRDR substitution appeared to influence the level of quinolone resistance expressed, and this varied with the quinolone agent examined. Close similarities occurred between the sequence and nature of QRDR changes in clinical and in vitro-generated quinolone-resistant mutants and with those previously reported for clinical and in vitro-generated quinolone-resistant gonococci. This suggests that quinolone resistance in meningococci may arise in the same manner and reach similar levels in vivo to those seen in quinolone-resistant Neisseria gonorrhoeae.

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