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Akiyo Nakano,Ryuichi Nakano,Yuki Suzuki,Kyoichi Saito,Kei Kasahara,Shiro Endo,Hisakazu Yano 대한진단검사의학회 2018 Annals of Laboratory Medicine Vol.38 No.4
Dear Editor, Carbapenem-resistant Enterobacteriaceae have acquired carbapenemase genes [1], which differ substantially across countries [2]. Transferable carbapenemase IMP-type metallo-β-lactamases, particularly IMP-1 and IMP-6, are commonly identified in the clinical setting in Japan [3, 4] and exhibit different substrate specificity despite having a difference of only one amino acid (IMP-6: Ser214Gly). IMP-1 producers are more resistant to imipenem than to meropenem, whereas IMP-6 producers are more resistant to meropenem [5]. We previously found that the susceptibility rate of IMP-6-positive Escherichia coli was higher for imipenem than for meropenem [3]. Thus, IMP-6-producing isolates may be erroneously categorized as imipenem-susceptible, which could lead to treatment failure in patients.
Naoki Kakuta,Ryuichi Nakano,Akiyo Nakano,Yuki Suzuki,Ayako Tanouchi,Takashi Masui,Saori Horiuchi,Shiro Endo,Risako Kakuta,Yasuo Ono,Hisakazu Yano 대한진단검사의학회 2020 Annals of Laboratory Medicine Vol.40 No.1
Background: Mutations in the quinolone resistance-determining regions (QRDRs) of Acinetobacter baumannii DNA gyrase (gyrA) and topoisomerase IV (parC) are linked to fluoroquinolone (FQ) resistance. We developed a mismatched PCR-restriction fragment length polymorphism (RFLP) assay to detect mutations in the gyrA and parC QRDRs associated with FQ resistance in A. baumannii. Methods: Based on the conserved sequences of A. baumannii gyrA and parC, two primer sets were designed for mismatched PCR-RFLP to detect mutations in gyrA (codons 83 and 87) and parC (codons 80 and 84) by introducing an artificial restriction enzyme cleavage site into the PCR products. This assay was evaluated using 58 A. baumannii strains and 37 other Acinetobacter strains that have been identified by RNA polymerase β-subunit gene sequence analysis. Results: PCR amplification of gyrA and parC was successful for all A. baumannii strains. In 11 FQ -susceptible strains, the gyrA and parC PCR products were digested by the selected restriction enzymes at the site containing gyrA (codons 83 and 87) and parC (codons 80 and 84). PCR products from 47 FQ-resistant strains containing mutations in gyrA and parC were not digested by the restriction enzymes at the site containing the mutation. As for the non-baumannii Acinetobacter strains, although amplification products for gyrA were obtained for 28 strains, no parC amplification product was obtained for any strain. Conclusions: This assay specifically amplified gyrA and parC from A. baumannii and detected A. baumannii gyrA and parC mutations with FQ resistance.