Brockmann Maria, Leineweber Christoph, Hellebuyck Tom, Martel An, Pasmans Frank, Gentil Michaela, Müller Elisabeth, Marschang Rachel E
Laboklin GmbH & Co. KG, Steubenstr. 4, 97688 Bad Kissingen, Germany.
Department of Pathobiology, Pharmacology and Zoological Medicine, Ghent University, Salisburylaan 133, B-9820 Merelbeke, Belgium.
Animals (Basel). 2023 Feb 28;13(5):881. doi: 10.3390/ani13050881.
(1) Background: () is a potential cause of dermatitis and cheilitis in lizards. The aim of this study was to establish a real-time PCR assay for the detection of . (2) Methods: Primers and probe were selected targeting the 16S rRNA gene, using sequences of 16S rRNA genes of as well as of other bacterial species derived from GenBank. The PCR assay was tested with 14 positive controls of different cultures as well as with 34 negative controls of various non- bacterial cultures. Additionally, samples of 38 lizards, mostly spp. and spp., submitted to a commercial veterinary laboratory were tested for the presence of using the established protocol. (3) Results: Concentrations of as low as 2 × 10 colonies per mL were detectable using dilutions of bacterial cell culture (corresponding to approximately 200 CFU per PCR). The assay resulted in an intraassay percent of coefficient of variation (CV) of 1.31% and an interassay CV of 1.80%. (4) Conclusions: The presented assay is able to detect in clinical samples, decreasing laboratory turn-around time in comparison to conventional culture-based detection methods.
(1) 背景：（某细菌名称未给出）是蜥蜴皮炎和唇炎的一个潜在病因。本研究的目的是建立一种用于检测（该细菌）的实时聚合酶链反应（PCR）检测方法。(2) 方法：利用从基因库获取的（该细菌）以及其他细菌物种的16S核糖体RNA（rRNA）基因序列，选择针对16S rRNA基因的引物和探针。用14种不同（该细菌）培养物的阳性对照以及34种各种非（该细菌）培养物的阴性对照对PCR检测方法进行测试。此外，使用既定方案对提交给一家商业兽医实验室的38只蜥蜴（大多为某蜥蜴属和另一蜥蜴属）的样本进行检测，以确定是否存在（该细菌）。(3) 结果：使用细菌细胞培养物稀释液可检测到低至每毫升2×10个菌落的浓度（相当于每个PCR约200个菌落形成单位）。该检测方法的批内变异系数（CV）百分比为1.31%，批间CV为1.80%。(4) 结论：所提出的检测方法能够在临床样本中检测到（该细菌），与传统的基于培养的检测方法相比，减少了实验室周转时间。
