Preziuso Silvia, Laus Fulvio, Tejeda Aurora Romero, Valente Carlo, Cuteri Vincenzo
Department of Veterinary Science, University of Camerino, Via Circonvallazione 93/95, Matelica, Italy.
J Vet Sci. 2010 Mar;11(1):67-72. doi: 10.4142/jvs.2010.11.1.67.
Streptococcus (S.) dysgalactiae subsp. equisimilis is responsible for severe diseases in humans, including primary bacteraemia, pneumonia, endocarditis, and toxic shock syndrome. Infection in some animal species can also occur, although a few studies have looked into cross-species infectivity. In horses, S. equisimilis is generally considered infrequent or opportunistic, but has recently been isolated from cases of strangles-like disease. Rapid and sensitive diagnostic techniques could enable epidemiological studies and effective investigation of outbreaks involving these bacteria. In this study, PCR protocols previously described in cattle and in humans to detect the species S. dysgalactiae and the subspecies equisimilis were evaluated to detect specific sequences in equine samples. For this purpose, 99 monolateral nasal swabs were collected from horses from stud farms with a history of S. equisimilis infection and were tested blindly by bacteriological isolation and by single and duplex PCR. DNA for PCR was extracted both from the colonies grown on agar media and from enrichment broth aliquots after incubation with nasal swab samples. S. equisimilis was identified by bacteriological isolation in 23 out of 99 swab samples, and PCR assays on these colonies were fully concordant with bacteriological identification (kappa statistic = 1.00). In addition, PCR of the enrichment broth aliquots confirmed the bacteriological results and detected S. equisimilis in 6 samples more than the bacteriological examination (kappa statistic = 0.84). The PCR protocols appeared to be reliable for the rapid identification of S. equisimilis in equine nasal swab samples, and could be useful for microbiological diagnosis.
马链球菌兽疫亚种可导致人类患上严重疾病,包括原发性菌血症、肺炎、心内膜炎和中毒性休克综合征。虽然有少数研究探讨了跨物种感染性,但在一些动物物种中也会发生感染。在马匹中,兽疫链球菌通常被认为不常见或属于机会致病菌,但最近已从类似马腺疫的病例中分离出来。快速且灵敏的诊断技术能够开展流行病学研究,并有效调查涉及这些细菌的疫情爆发情况。在本研究中,对先前在牛和人类中描述的用于检测马链球菌兽疫亚种的PCR方案进行了评估,以检测马样本中的特定序列。为此,从有兽疫链球菌感染病史的种马场的马匹中采集了99份单侧鼻拭子,并通过细菌学分离以及单重和双重PCR进行盲法检测。用于PCR的DNA既从琼脂培养基上生长的菌落中提取,也从与鼻拭子样本孵育后的富集肉汤等分试样中提取。在99份拭子样本中,通过细菌学分离鉴定出23份含有兽疫链球菌,对这些菌落进行的PCR检测结果与细菌学鉴定完全一致(kappa统计量 = 1.00)。此外,对富集肉汤等分试样进行的PCR证实了细菌学检测结果,并且比细菌学检测多检测出6份含有兽疫链球菌的样本(kappa统计量 = 0.84)。这些PCR方案对于快速鉴定马鼻拭子样本中的兽疫链球菌似乎是可靠的,并且可能有助于微生物学诊断。