Carlos Bastidas-Caldes, Fernanda Hernández-Alomía, Miguel Almeida, Mirian Ormaza, Josué Boada, Jay Graham, Manuel Calvopiña, Pablo Castillejo
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In this study, we aimed to re-identifying Enterobacterales as the primary bacterial agents responsible for urinary tract infections (UTIs) by comparing the sensitivity and specificity of traditional microbiological methods with advanced molecular techniques for the detection of uropathogens in indigenous women from Otavalo, Ecuador.</p><p><strong>Methods: </strong>A facility-based cross-sectional study was conducted from October 2021 to February 2022 among Kichwa-Otavalo women. Pathogens from urine samples were identified using culture and biochemical typing. Morphological identification was doble-checked through PCR and DNA sequencing of 16S, recA, and rpoB molecular barcodes. The isolates were subjected to antimicrobial susceptibility-testing using disk diffusion test.</p><p><strong>Results: </strong>This study highlighted a 32% misidentification rate between biochemical and molecular identification. Using traditional methods, E. coli was 26.19% underrepresented meanwhile Klebsiella oxytoca was overrepresented by 92.86%. Furthermore, the genera Pseudomonas, Proteus, and Serratia were confirmed to be E. coli and Klebsiella spp. by molecular method, and one Klebsiella spp. was reidentified as Enterobacter spp. The susceptibility profile showed that 59% of the isolates were multidrug resistant strains and 31% produced extended spectrum beta-lactamases (ESBLs). Co-trimoxazole was the least effective antibiotic with 61% of the isolates resistant. Compared to previous reports, resistance to nitrofurantoin and fosfomycin showed an increase in resistance by 25% and 15%, respectively.</p><p><strong>Conclusions: </strong>Community-acquired UTIs in indigenous women in Otavalo were primarily caused by E. coli and Klebsiella spp. Molecular identification (16S/rpoB/recA) revealed a high rate of misidentification by standard biochemical and microbiological techniques, which could lead to incorrect antibiotic prescriptions. UTI isolates in this population displayed higher levels of resistance to commonly used antibiotics compared with non-indigenous groups. Accurate identification of pathogens causing UTIs and their antibiotic susceptibility in local populations is important for local antibiotic prescribing guidelines.</p>","PeriodicalId":8981,"journal":{"name":"BMC Infectious Diseases","volume":null,"pages":null},"PeriodicalIF":3.4000,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11515717/pdf/","citationCount":"0","resultStr":"{\"title\":\"Molecular identification and antimicrobial resistance patterns of enterobacterales in community urinary tract infections among indigenous women in Ecuador: addressing microbiological misidentification.\",\"authors\":\"Carlos Bastidas-Caldes, Fernanda Hernández-Alomía, Miguel Almeida, Mirian Ormaza, Josué Boada, Jay Graham, Manuel Calvopiña, Pablo Castillejo\",\"doi\":\"10.1186/s12879-024-10096-7\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>Antibiotic resistance of Enterobacterales poses a major challenge in the treatment of urinary tract infections (UTIs). In low- and middle-income countries (LMICs), standard microbiological (i.e. urine culture and simple disk diffusion test) methods are considered the \\\"gold standard\\\" for bacterial identification and drug susceptibility testing, while PCR and DNA sequencing are less commonly used. In this study, we aimed to re-identifying Enterobacterales as the primary bacterial agents responsible for urinary tract infections (UTIs) by comparing the sensitivity and specificity of traditional microbiological methods with advanced molecular techniques for the detection of uropathogens in indigenous women from Otavalo, Ecuador.</p><p><strong>Methods: </strong>A facility-based cross-sectional study was conducted from October 2021 to February 2022 among Kichwa-Otavalo women. Pathogens from urine samples were identified using culture and biochemical typing. Morphological identification was doble-checked through PCR and DNA sequencing of 16S, recA, and rpoB molecular barcodes. The isolates were subjected to antimicrobial susceptibility-testing using disk diffusion test.</p><p><strong>Results: </strong>This study highlighted a 32% misidentification rate between biochemical and molecular identification. Using traditional methods, E. coli was 26.19% underrepresented meanwhile Klebsiella oxytoca was overrepresented by 92.86%. Furthermore, the genera Pseudomonas, Proteus, and Serratia were confirmed to be E. coli and Klebsiella spp. by molecular method, and one Klebsiella spp. was reidentified as Enterobacter spp. The susceptibility profile showed that 59% of the isolates were multidrug resistant strains and 31% produced extended spectrum beta-lactamases (ESBLs). Co-trimoxazole was the least effective antibiotic with 61% of the isolates resistant. 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引用次数: 0
摘要
背景:肠杆菌的抗生素耐药性是治疗尿路感染(UTI)的一大挑战。在中低收入国家(LMICs),标准微生物学方法(即尿液培养和简单的磁盘扩散试验)被认为是细菌鉴定和药敏试验的 "金标准",而 PCR 和 DNA 测序则较少使用。在这项研究中,我们旨在通过比较传统微生物学方法和先进分子技术在厄瓜多尔奥塔瓦洛土著妇女尿路病原体检测中的灵敏度和特异性,重新确定肠杆菌科细菌是导致尿路感染(UTI)的主要细菌:方法:2021 年 10 月至 2022 年 2 月,在 Kichwa-Otavalo 妇女中开展了一项基于设施的横断面研究。通过培养和生化分型鉴定了尿液样本中的病原体。通过对 16S、recA 和 rpoB 分子条形码进行 PCR 和 DNA 测序,对形态学鉴定进行双重检查。利用盘扩散试验对分离物进行抗菌药敏感性测试:结果:这项研究表明,生化鉴定和分子鉴定之间的误判率高达 32%。使用传统方法,大肠杆菌的比例偏低 26.19%,而土生克雷伯菌的比例偏高 92.86%。此外,假单胞菌属、变形杆菌属和沙雷氏菌属通过分子方法被确认为大肠杆菌和克雷伯菌属,其中一个克雷伯菌属被重新鉴定为肠杆菌属。 药敏谱显示,59%的分离菌株为多重耐药菌株,31%产生广谱β-内酰胺酶(ESBLs)。共三唑是效果最差的抗生素,61%的分离菌株对其产生耐药性。与之前的报告相比,对硝基呋喃妥因和磷霉素的耐药性分别增加了25%和15%:分子鉴定(16S/rpoB/recA)显示,标准生化和微生物技术的错误鉴定率很高,这可能导致错误的抗生素处方。与非土著群体相比,该人群中的 UTI 分离物对常用抗生素的耐药性水平更高。准确鉴定当地人群中导致UTI的病原体及其对抗生素的敏感性对于制定当地抗生素处方指南非常重要。
Molecular identification and antimicrobial resistance patterns of enterobacterales in community urinary tract infections among indigenous women in Ecuador: addressing microbiological misidentification.
Background: Antibiotic resistance of Enterobacterales poses a major challenge in the treatment of urinary tract infections (UTIs). In low- and middle-income countries (LMICs), standard microbiological (i.e. urine culture and simple disk diffusion test) methods are considered the "gold standard" for bacterial identification and drug susceptibility testing, while PCR and DNA sequencing are less commonly used. In this study, we aimed to re-identifying Enterobacterales as the primary bacterial agents responsible for urinary tract infections (UTIs) by comparing the sensitivity and specificity of traditional microbiological methods with advanced molecular techniques for the detection of uropathogens in indigenous women from Otavalo, Ecuador.
Methods: A facility-based cross-sectional study was conducted from October 2021 to February 2022 among Kichwa-Otavalo women. Pathogens from urine samples were identified using culture and biochemical typing. Morphological identification was doble-checked through PCR and DNA sequencing of 16S, recA, and rpoB molecular barcodes. The isolates were subjected to antimicrobial susceptibility-testing using disk diffusion test.
Results: This study highlighted a 32% misidentification rate between biochemical and molecular identification. Using traditional methods, E. coli was 26.19% underrepresented meanwhile Klebsiella oxytoca was overrepresented by 92.86%. Furthermore, the genera Pseudomonas, Proteus, and Serratia were confirmed to be E. coli and Klebsiella spp. by molecular method, and one Klebsiella spp. was reidentified as Enterobacter spp. The susceptibility profile showed that 59% of the isolates were multidrug resistant strains and 31% produced extended spectrum beta-lactamases (ESBLs). Co-trimoxazole was the least effective antibiotic with 61% of the isolates resistant. Compared to previous reports, resistance to nitrofurantoin and fosfomycin showed an increase in resistance by 25% and 15%, respectively.
Conclusions: Community-acquired UTIs in indigenous women in Otavalo were primarily caused by E. coli and Klebsiella spp. Molecular identification (16S/rpoB/recA) revealed a high rate of misidentification by standard biochemical and microbiological techniques, which could lead to incorrect antibiotic prescriptions. UTI isolates in this population displayed higher levels of resistance to commonly used antibiotics compared with non-indigenous groups. Accurate identification of pathogens causing UTIs and their antibiotic susceptibility in local populations is important for local antibiotic prescribing guidelines.
期刊介绍:
BMC Infectious Diseases is an open access, peer-reviewed journal that considers articles on all aspects of the prevention, diagnosis and management of infectious and sexually transmitted diseases in humans, as well as related molecular genetics, pathophysiology, and epidemiology.