Pinthanon Artitaya, Nithitarnwat Chayanan, Pintapin Chadaporn, Siripanee Chonradee, Yindee Jitrapa, Am-In Nutthee, Kesdangsakonwut Sawang, Surachetpong Sirilak, Prapasarakul Nuvee
Department of Veterinary Microbiology, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Department of Veterinary Obstetrics, Gynaecology and Reproduction, Faculty of Veterinary Science, Chulalongkorn University, Bangkok, 10330, Thailand.
Vet Res Commun. 2023 Sep;47(3):1457-1469. doi: 10.1007/s11259-023-10096-z. Epub 2023 Apr 10.
Using the matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) method for bacterial diagnosis, rapid urine sample preparation can reduce time relapsing of diagnosis and improve discriminatory power in coinfection cases. We aimed to evaluate rapid urine preparation procedures before MALDI-TOF MS application using dog clinical urine samples in comparison with standard microbiological diagnostic methods by agreement analysis. We determined the frequency and distribution of bacteria and bacterial resistance and their correlations to clinical history. Three experimental procedures comprising direct centrifugation, 10% sodium dodecyl sulfate digestion, and ultrasonic preparation were performed for method validation and sensitivity. Sterile urine containing Escherichia coli and/or Staphylococcus aureus were used as simulated samples. By ultrasonic preparation, the microorganisms could be detected 1.46-1.51 × 10 CFU, which was considered the most suitable technique. This preparation was significantly consistent with the routine method based on data from Hospital Information Systems for 50 urine samples from canine cystitis. By standard protocol, Enterobacteriaceae and Staphylococcus pseudintermedius were found in most of the 155 urine samples with cystitis. Extended-spectrum beta-lactamase-producing Enterobacteriaceae was found in 25-30% of the samples. Imipenem resistance was found in 70% of Acinetobacter baumannii cases; almost all were resistant to second-generation fluoroquinolones and tetracyclines. The most efficient antibiotic for treating bacterial urinary tract infection was amoxicillin plus clavulanic acid. A. baumannii and Pseudomonas aeruginosa were susceptible to pradofloxacin. Prolonged urine catheterization was linked to lower urinary tract infections by Enterobacter spp., which also correlated with chronic kidney disease.
使用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）方法进行细菌诊断时，快速的尿液样本制备可减少诊断的时间延误，并提高混合感染病例中的鉴别能力。我们旨在通过一致性分析，评估在使用犬类临床尿液样本进行MALDI-TOF MS检测之前的快速尿液制备程序，并与标准微生物诊断方法进行比较。我们确定了细菌的频率和分布、细菌耐药性及其与临床病史的相关性。为了进行方法验证和敏感性评估，我们进行了三种实验程序，包括直接离心、10%十二烷基硫酸钠消化和超声制备。含有大肠杆菌和/或金黄色葡萄球菌的无菌尿液用作模拟样本。通过超声制备，可以检测到1.46-1.51×10 CFU的微生物，这被认为是最合适的技术。根据医院信息系统中50份犬膀胱炎尿液样本的数据，这种制备方法与常规方法具有显著的一致性。按照标准方案，在155份膀胱炎尿液样本中，大多数样本检测到肠杆菌科细菌和中间型葡萄球菌。在25%-30%的样本中发现了产超广谱β-内酰胺酶的肠杆菌科细菌。在70%的鲍曼不动杆菌病例中发现了亚胺培南耐药；几乎所有菌株都对第二代氟喹诺酮类药物和四环素耐药。治疗细菌性尿路感染最有效的抗生素是阿莫西林加克拉维酸。鲍曼不动杆菌和铜绿假单胞菌对普拉氟沙星敏感。长时间留置导尿管与肠杆菌属引起的下尿路感染有关，这也与慢性肾病相关。
