Boyle Ashley G, Stefanovski Darko, Rankin Shelley C
Department of Clinical Studies New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, 382 West Street Rd, Kennett Square, PA, 19348, USA.
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 3900 Spruce Street, Room 4036, Philadelphia, PA, 19104, USA.
BMC Vet Res. 2017 Mar 23;13(1):75. doi: 10.1186/s12917-017-0989-4.
Streptococcus equi subsp equi (S. equi) is the cause of "equine strangles" which is a highly infectious upper respiratory disease. Detection of S. equi is influenced by site of specimen collection, method of sampling, and type of diagnostic test that is performed. We hypothesized i) that a loop-mediated isothermal amplification (LAMP) assay that targets the S. equi-specific eqbE gene would be more sensitive than a realtime PCR assay that targets the S. equi-specific seeI gene and ii) that LAMP of specimens obtained by guttural pouch lavage (GPL) would be more sensitive than LAMP of nasopharyngeal specimens to identify S. equi carriers.
A nasopharyngeal flocked swab, nasopharyngeal wash, and GPL specimen was collected from 44 convalescent horses and the eqbE LAMP assay was performed. The seeI realtime PCR assay and aerobic culture were also performed on the GPL specimen. Logistic regression was performed to compare sampling sites and test methods (P-values ≤0.05 were considered significant).
One of 41 nasopharyngeal flocked swabs, 6/38 nasopharyngeal wash and 24/44 GPL specimens were positive by eqbE LAMP. 18/44 GPL specimens were positive by seeI PCR and S. equi was isolated from 4/44 of these specimens. Detection of S. equi DNA was 51 times more likely from the GPL samples than nasopharyngeal samples (OR 51.0, P < 0.0001). When eqbE LAMP GPL samples were positive, it was eight times more likely that the guttural pouch had any abnormality on endoscopy (OR 8.2, P ≤ 0.005), almost 20 times more likely that mild empyema was found (OR 19.7, P ≤ 0.002), and eight times more likely that the SeeI PCR was positive for S. equi DNA (OR 8.1, P ≤ 0.006).
This study demonstrates that guttural pouch lavage specimens should be used to detect S. equi and that the eqbE LAMP assay was comparable to the seeI PCR.
马链球菌兽疫亚种(S. equi)是“马腺疫”的病原体,这是一种具有高度传染性的上呼吸道疾病。S. equi的检测受标本采集部位、采样方法和所进行的诊断测试类型的影响。我们假设:i)靶向S. equi特异性eqbE基因的环介导等温扩增(LAMP)检测比靶向S. equi特异性seeI基因的实时PCR检测更灵敏;ii)通过咽鼓管囊冲洗(GPL)获得的标本进行LAMP检测比鼻咽标本的LAMP检测在识别S. equi携带者方面更灵敏。
从44匹康复马中采集鼻咽植绒拭子、鼻咽冲洗液和GPL标本,并进行eqbE LAMP检测。对GPL标本还进行了seeI实时PCR检测和好氧培养。进行逻辑回归以比较采样部位和检测方法(P值≤0.05被认为具有显著性)。
41份鼻咽植绒拭子中有1份、38份鼻咽冲洗液中有6份以及44份GPL标本中有24份通过eqbE LAMP检测呈阳性。44份GPL标本中有18份通过seeI PCR检测呈阳性,并且从这些标本中的4份分离出了S. equi。检测到S. equi DNA的GPL样本比鼻咽样本的可能性高51倍(比值比51.0,P<0.0001)。当eqbE LAMP GPL样本呈阳性时,咽鼓管囊在内镜检查时有任何异常的可能性高8倍(比值比8.2,P≤0.005),发现轻度脓胸的可能性高近20倍(比值比19.7,P≤0.002),并且seeI PCR检测S. equi DNA呈阳性的可能性高8倍(比值比8.1,P≤0.006)。
本研究表明应使用咽鼓管囊冲洗标本检测S. equi,并且eqbE LAMP检测与seeI PCR相当。