Gupta Ruchi, Gaind Rajni, Singh Laishram Chandreshwor, Paglietti Bianca, Deb Monorama, Rubino Salvatore, Wain John, Basir Seemi Farhat
Department of Biosciences, Jamia Millia Islamia, New Delhi 110025, India.
Department of Microbiology, Vardhman Mahavir Medical College and Safdarjung Hospital, New Delhi 110029, India.
Trans R Soc Trop Med Hyg. 2016 Dec 1;110(12):684-689. doi: 10.1093/trstmh/trx002.
Fluoroquinolone resistance is mediated by mutations in the quinolone-resistance determining region (QRDR) of the topoisomerase genes. Denaturing high performance liquid chromatography (DHPLC) was evaluated for detection of clinically important mutations in gyrB among Salmonella.
Salmonella Typhi and S. Paratyphi A characterised for mutation in QRDR of gyrA, parC and parE were studied for mutation in gyrB by DHPLC and validated by sequencing.
The DHPLC analysis was able to resolve the test mutant from isolates with wild type gyrB and distinguished mutants from other mutant by peak profile and shift in retention time. Three sequence variants were detected at codon 464, and a novel mutation Ser→Thr was also detected. gyrB mutation was associated with non classical quinolone resistance (NALS-CIPDS) in 34 isolates of S. Typhi only and was distinct from classical quinolone resistance associated with gyrA mutations (NALR-CIPDS).
DHPLC is effective for the detection of mutation and can reduce the need for sequencing to detect clinically significant gyrB mutations.
KF993966, KF993965 and KF993964.
氟喹诺酮耐药性由拓扑异构酶基因喹诺酮耐药决定区（QRDR）的突变介导。评估了变性高效液相色谱法（DHPLC）用于检测沙门氏菌中gyrB基因临床上重要的突变。
对gyrA、parC和parE的QRDR突变特征明确的伤寒沙门氏菌和甲型副伤寒沙门氏菌，通过DHPLC研究gyrB基因的突变，并通过测序进行验证。
DHPLC分析能够将测试突变体与具有野生型gyrB的分离株区分开来，并通过峰型和保留时间的变化将突变体与其他突变体区分开来。在第464密码子处检测到三个序列变体，还检测到一个新的突变Ser→Thr。gyrB突变仅与34株伤寒沙门氏菌中的非经典喹诺酮耐药性（NALS-CIPDS）相关，且与与gyrA突变相关的经典喹诺酮耐药性（NALR-CIPDS）不同。
DHPLC在检测突变方面有效，可减少对测序的需求以检测临床上显著的gyrB突变。
KF993966、KF993965和KF993964。
