Soliman Ahmed M, Nariya Hirofumi, Tanaka Daiki, Shimamoto Toshi, Shimamoto Tadashi
Department of Microbiology and Immunology, Faculty of Pharmacy, Kafrelsheikh University, Kafr El-Sheikh 33516, Egypt.
Laboratory of Food Microbiology, Graduate School of Human Life Sciences, Jumonji University, Niiza 352-8510, Japan.
Antibiotics (Basel). 2023 Jan 4;12(1):90. doi: 10.3390/antibiotics12010090.
We developed two multiplex polymerase chain reactions (PCRs) for the detection of extended-spectrum β-lactamases (ESBLs), plasmid-mediated AmpC β-lactamases, gene, and integrase genes (, , and ) in class 1, 2, and 3 integrons in Gram-negative bacteria. We evaluated the PCRs using 109 Gram-negative isolates from non-organic (ANO) and organic (AO) vegetables and fruits. Screening of ANO substances identified five SHV, one TEM-1, one CTX-M, 20 AmpC-CS, and two positives. DNA sequencing revealed CTX-M in spp. was , a plasmid-mediated CTX-M related ESBL gene only found in spp. Of the 20 AmpC-CS positives, 10 were CMY/MIR/ACT/EC (3 new variants), eight were ACT, one was AZECL, and one was new -related AmpC family. Screening of AO substances identified 11 SHV, two TEM-1, three CTX-M (one OXY-2, two CTX-M-14/-15), two OXA-9, 13 AmpC-CS and one positives. The 13 AmpC-CS positives were five CMY/MIR/ACT/EC, three ACT, one MOX-12 variant, and four ADC (one ADC-25 and three new variants). We developed a rapid, easy-to-perform, low-cost, and reliable multiplex PCR system for screening clinically relevant β-lactamases and integrons in Gram-negative bacteria. We showed the prevalence of ESBLs and AmpC β-lactamases among our panel of ampicillin-resistant Gram-negative strains and detection of NDM and OXA carbapenemases.
我们开发了两种多重聚合酶链反应（PCR），用于检测革兰氏阴性菌中1类、2类和3类整合子中的超广谱β-内酰胺酶（ESBLs）、质粒介导的AmpCβ-内酰胺酶、基因和整合酶基因（、和）。我们使用从非有机（ANO）蔬菜和水果以及有机（AO）蔬菜和水果中分离出的109株革兰氏阴性菌对这些PCR进行了评估。对ANO物质的筛查发现了5种SHV、1种TEM-1、1种CTX-M、20种AmpC-CS以及2种阳性。DNA测序显示，在 spp. 中的CTX-M是，这是一种仅在 spp. 中发现的质粒介导的与CTX-M相关的ESBL基因。在20种AmpC-CS阳性菌株中，10种是CMY/MIR/ACT/EC（3种新变体），8种是ACT，1种是AZECL，1种是新的 -相关AmpC家族。对AO物质的筛查发现了11种SHV、2种TEM-1、3种CTX-M（1种OXY-2、2种CTX-M-14/-15）、2种OXA-9、13种AmpC-CS以及1种阳性。这13种AmpC-CS阳性菌株分别为5种CMY/MIR/ACT/EC、3种ACT、1种MOX-12变体和4种ADC（1种ADC-25和3种新变体）。我们开发了一种快速、易于操作、低成本且可靠的多重PCR系统，用于筛查革兰氏阴性菌中临床相关的β-内酰胺酶和整合子。我们展示了在我们的耐氨苄西林革兰氏阴性菌株组中ESBLs和AmpCβ-内酰胺酶的流行情况以及NDM和OXA碳青霉烯酶的检测情况。
