Integrated Biosensor Assay for Rapid Uropathogen Identification and Phenotypic Antimicrobial Susceptibility Testing.
Integrated Biosensor Assay for Rapid Uropathogen Identification and Phenotypic Antimicrobial Susceptibility Testing.
复制标题
DOI：
10.1016/j.euf.2015.12.010
复制
发表时间：
2017-04
影响因子：
5.4
通讯作者：
Liao JC
中科院分区：
文献类型：
--
作者：
Altobelli E;Mohan R;Mach KE;Sin MLY;Anikst V;Buscarini M;Wong PK;Gau V;Banaei N;Liao JC
通过尿培养进行病原体鉴定（ID）和抗菌药物敏感性测试（AST）的尿路感染（UTI）的标准诊断需要2-3天。这种延误导致经验性治疗，并导致滥用抗生素和耐药病原体的增加。UTI的快速诊断测试可以改善患者护理和抗生素管理。开发和验证用于UTI诊断的集成生物传感器测定，包括病原体ID和AST，并测定环丙沙星的最小抑菌浓度（MIC）。2013年11月至2014年9月期间，从斯坦福大学临床微生物实验室获得肠杆菌科阳性（n = 84）或培养阴性（n = 23）的尿液样本。将每个样品稀释并在有和没有环丙沙星的情况下培养5 h，然后使用用一组互补DNA探针功能化的单个电化学生物传感器阵列定量检测细菌16 S rRNA。使用通用细菌、肠杆菌科（EB）和病原体特异性探针确定病原体ID。使用EB探针测定表型AST与环丙沙星MIC，以测量16 S rRNA水平作为细菌生长的函数。在6SD处超过背景的病原体ID的电化学信号被认为是阳性。比无抗生素对照低0.4 log单位的MIC信号指示敏感性。将结果与临床微生物学报告进行比较。对于病原体ID，该测定具有98.5%的灵敏度、96.6%的特异性、93.0%的阳性预测值和99.3%的阴性预测值。对于环丙沙星MIC，分类和基本一致性为97.6%。进一步的自动化、额外病原体和抗生素的测试以及全面的前瞻性研究对于转化为临床应用是必要的。集成的生物传感器平台在显著更短的测定时间内实现了与标准培养物相当的微生物学结果，包括MIC。进一步的测定自动化将允许临床翻译用于UTI的快速分子诊断。我们已经开发并验证了一种用于快速诊断尿路感染的生物传感器测试。该器械的临床转化有可能显著加快和改善尿路感染的治疗。
Standard diagnosis of urinary tract infection (UTI) via urine culture for pathogen identification (ID) and antimicrobial susceptibility testing (AST) takes 2–3 d. This delay results in empiric treatment and contributes to the misuse of antibiotics and the rise of resistant pathogens. A rapid diagnostic test for UTI may improve patient care and antibiotic stewardship. To develop and validate an integrated biosensor assay for UTI diagnosis, including pathogen ID and AST, with determination of the minimum inhibitory concentration (MIC) for ciprofloxacin. Urine samples positive for Enterobacteriaceae (n = 84) or culture-negative (n = 23) were obtained from the Stanford Clinical Microbiology Laboratory between November 2013 and September 2014. Each sample was diluted and cultured for 5 h with and without ciprofloxacin, followed by quantitative detection of bacterial 16S rRNA using a single electrochemical biosensor array functionalized with a panel of complementary DNA probes. Pathogen ID was determined using universal bacterial, Enterobacteriaceae (EB), and pathogen-specific probes. Phenotypic AST with ciprofloxacin MIC was determined using an EB probe to measure 16S rRNA levels as a function of bacterial growth. Electrochemical signals for pathogen ID at 6 SD over background were considered positive. An MIC signal of 0.4 log units lower than the no-antibiotic control indicated sensitivity. Results were compared to clinical microbiology reports. For pathogen ID, the assay had 98.5% sensitivity, 96.6% specificity, 93.0% positive predictive value, and 99.3% negative predictive value. For ciprofloxacin MIC the categorical and essential agreement was 97.6%. Further automation, testing of additional pathogens and antibiotics, and a full prospective study will be necessary for translation to clinical use. The integrated biosensor platform achieved microbiological results including MIC comparable to standard culture in a significantly shorter assay time. Further assay automation will allow clinical translation for rapid molecular diagnosis of UTI. We have developed and validated a biosensor test for rapid diagnosis of urinary tract infections. Clinical translation of this device has the potential to significantly expedite and improve treatment of urinary tract infections.
登录
查看更多内容
影响因子：
11.8
作者：
Gupta, Kalpana;Hooton, Thomas M.;Soper, David E.
通讯作者：
Soper, David E.
影响因子：
13.8
作者：
Mach KE;Wong PK;Liao JC
通讯作者：
Liao JC
影响因子：
9.4
作者：
Liao, JC;Mastali, M;Haake, DA
通讯作者：
Haake, DA
影响因子：
2.2
作者：
Wang, X. -H.;Zhang, G.;Lu, X. -X.
通讯作者：
Lu, X. -X.
影响因子：
3.7
作者：
Burillo A;Rodríguez-Sánchez B;Ramiro A;Cercenado E;Rodríguez-Créixems M;Bouza E
通讯作者：
Bouza E