Patty O A, Cursons R T M
a Biological Sciences , The University of Waikato , Private Bag 3105, Hamilton 3240 , New Zealand.
N Z Vet J. 2014 Mar;62(2):63-7. doi: 10.1080/00480169.2013.841536. Epub 2013 Oct 24.
To identify Streptococcus equi subsp. equi (S. equi) by PCR analysis and obtain isolates by culture, in order to investigate the strains of S. equi infecting horses within New Zealand.
A diagnostic PCR, based on the amplification of the seeI gene for S. equi, was used on 168 samples submitted from horses with and without clinical signs of strangles. Samples were also processed and cultured on selective media for the isolation of β-haemolytic colonies. In addition, the hypervariable region of the seM gene of S. equi was amplified and then sequenced for strain typing purposes.
Of the 168 samples, 35 tested positive for S. equi using PCR. Thirty-two confirmed samples were from horses with a clinical diagnosis of strangles and three were from horses where clinical information was unavailable. Only 22/35 (63%) confirmed S. equi samples were successfully isolated following culture. Strain typing demonstrated that two novel seM alleles of S. equi were found in New Zealand with SeM-99 strains being restricted to the North Island while SeM-100 strains were found in both North and South Islands.
The application of PCR for the laboratory confirmation of strangles allowed for a rapid and sensitive identification of S. equi. Moreover, seM typing revealed that within the samples examined two strains of S. equi co-circulated within the North Island of New Zealand but only one strain in the South Island.
PCR reduces the time required to obtain laboratory confirmation of strangles compared with culture methods. It also has greater sensitivity in detecting S. equi infections, which is of particular importance in the detection of carrier animals which normally shed low numbers of bacteria. Additionally, seM molecular typing can differentiate between bacterial strains, assisting in the monitoring of local strains of S. equi subsp. equi causing disease.
通过聚合酶链反应(PCR)分析鉴定马链球菌兽疫亚种(S. equi),并通过培养获得分离株,以调查感染新西兰马匹的S. equi菌株。
基于扩增S. equi的seeI基因进行诊断性PCR,对168份有或无腺疫临床症状的马匹提交的样本进行检测。样本还在选择性培养基上进行处理和培养,以分离β-溶血菌落。此外,扩增S. equi的seM基因高变区,然后进行测序以进行菌株分型。
在168份样本中,35份通过PCR检测为S. equi阳性。32份确诊样本来自临床诊断为腺疫的马匹,3份来自临床信息不详的马匹。培养后仅22/35(63%)的确诊S. equi样本成功分离。菌株分型表明,在新西兰发现了两个新的S. equi seM等位基因,SeM-99菌株仅限于北岛,而SeM-100菌株在北岛和南岛均有发现。
PCR用于腺疫的实验室确诊,可快速、灵敏地鉴定S. equi。此外,seM分型显示,在所检测的样本中,两种S. equi菌株在新西兰北岛共同流行,但在南岛只有一种菌株。
与培养方法相比,PCR减少了获得腺疫实验室确诊所需的时间。它在检测S. equi感染方面也具有更高的灵敏度,这在检测通常排出少量细菌的携带动物时尤为重要。此外,seM分子分型可以区分细菌菌株,有助于监测引起疾病的马链球菌兽疫亚种的本地菌株。