Lund Heidi Sjetne, Skogtun Gaute, Sørum Henning, Eggertsdóttir Anna Vigdís
Department of Companion Animal Clinical Sciences, Norwegian University of Life Sciences, Oslo, Norway
Department of Food Safety and Infection Biology, Norwegian University of Life Sciences, Oslo, Norway.
J Feline Med Surg. 2015 Oct;17(10):909-14. doi: 10.1177/1098612X14563098. Epub 2014 Dec 22.
A diagnosis of bacterial cystitis commonly relies on a positive microbiological culture demonstrating the presence of a significant number of colony-forming units/ml urine, as urine within the upper urinary tract, bladder and proximal urethra generally is considered sterile. Recent studies from human and veterinary medicine indicate the presence of non-culturable bacteria in culture-negative urine samples. The aim of the present study was to determine the occurrence of bacterial DNA in culture-negative urine samples from cats with signs of feline lower urinary tract disease (FLUTD) and healthy control cats by 16S ribosomal DNA PCR and subsequent sequencing. The study sample included 38 culture-negative urine samples from cats with FLUTD and 43 culture-negative samples from control cats. Eight culture-positive urine samples from cats with FLUTD were included as external positive controls in addition to negative reaction controls. Of possible methodological limitations, degradation of DNA due to storage, the use of non-sedimented urine for DNA isolation and lack of internal positive reaction controls should be mentioned. The positive controls were recognised, but occurrence of bacterial DNA in culture-negative urine from cats with or without signs of lower urinary tract disease was not demonstrated. However, considering the possible methodological limitations, the presence of bacterial DNA in the urine of culture-negative FLUTD cats cannot be excluded based on the present results alone. Therefore, a prospective study reducing the possibility of degradation of DNA due to storage, in combination with modifications enhancing the chance of detecting even lower levels of bacterial DNA in culture-negative samples, seems warranted.
细菌性膀胱炎的诊断通常依赖于微生物培养呈阳性,即每毫升尿液中存在大量菌落形成单位,因为上尿路、膀胱和近端尿道内的尿液通常被认为是无菌的。来自人类和兽医学的最新研究表明,在培养阴性的尿液样本中存在不可培养细菌。本研究的目的是通过16S核糖体DNA聚合酶链反应(PCR)及后续测序,确定患有猫下泌尿道疾病(FLUTD)体征的猫和健康对照猫的培养阴性尿液样本中细菌DNA的存在情况。研究样本包括38份来自患有FLUTD的猫的培养阴性尿液样本和43份来自对照猫的培养阴性样本。除阴性反应对照外,还包括8份来自患有FLUTD的猫的培养阳性尿液样本作为外部阳性对照。在可能的方法学局限性方面,应提及由于储存导致的DNA降解、使用未沉淀尿液进行DNA分离以及缺乏内部阳性反应对照。阳性对照得到了识别,但未证实患有或未患有下泌尿道疾病体征的猫的培养阴性尿液中存在细菌DNA。然而,考虑到可能的方法学局限性,仅凭目前的结果不能排除培养阴性的FLUTD猫尿液中存在细菌DNA的可能性。因此,开展一项前瞻性研究,减少由于储存导致DNA降解的可能性,并结合改进措施以提高在培养阴性样本中检测到更低水平细菌DNA的机会,似乎是有必要的。