Rapid Visual Detection of Highly Pathogenic Streptococcus suis Serotype 2 Isolates by Use of Loop-Mediated Isothermal Amplification
Rapid Visual Detection of Highly Pathogenic Streptococcus suis Serotype 2 Isolates by Use of Loop-Mediated Isothermal Amplification
复制标题
使用环介导等温扩增快速视觉检测高致病性猪链球菌血清型 2 分离株
DOI：
10.1128/jcm.01183-13
复制
发表时间：
2013-10-01
影响因子：
9.4
通讯作者：
Qia, Zhongtian
中科院分区：
文献类型：
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作者：
Zhang, Jinhai;Zhu, Jing;Qia, Zhongtian
猪链球菌2型（S.猪流感是一种重要的人畜共患病原体，给养猪业造成巨大的经济损失，并严重威胁全球公众健康。高致病性S.含有89 K致病岛（派）的suis 2已在人类中引起大规模感染暴发，导致高死亡率。在这项研究中，我们建立了两种基于环介导等温扩增（LAMP）的方法，可以快速检测S。suis 2和89 K派，并且可以在相同条件下同时进行。此外，基于这两种LAMP检测的结果，并使用同一组连续稀释的DNA样品，我们比较了不同LAMP产物检测方法的灵敏度，包括SYBR绿色检测、凝胶电泳、比浊法、钙黄绿素检测和羟基萘酚蓝检测。结果表明，在63 ℃恒温条件下，48 min内可扩增和检测目的基因。S. suis 2的检测因检测方法和反应体系而异，表明对于每种LAMP反应体系，应进行多种检测方法以选择最佳方法。优化后的LAMP方法的灵敏度（7.16拷贝/反应）与实时荧光PCR方法的灵敏度相当，对参考菌株和临床样本的检测结果表明，该LAMP系统具有较高的特异性。因此，由于本研究建立的LAMP系统是简单，快速，敏感，他们可能有很好的临床潜力，用于检测高致病性S。suis 2.
Streptococcus suis serotype 2 (S. suis 2) is an important zoonotic pathogen that causes considerable economic losses to the pig industry and significantly threatens public health worldwide. The highly pathogenic S. suis 2, which contains the 89K pathogenicity island (PAI), has caused large-scale outbreaks of infections in humans, resulting in high mortality rates. In this study, we established two loop-mediated isothermal amplification (LAMP)-based assays that can rapidly detect S. suis 2 and the 89K PAI and can be performed simultaneously under the same conditions. Further, based on the findings of these two LAMP assays and using the same set of serially diluted DNA samples, we compared the sensitivities of different LAMP product detection methods, including SYBR green detection, gel electrophoresis, turbidimetry, calcein assays, and hydroxynaphthol blue detection. The results suggest that target genes can be amplified and detected within 48 min under 63 degrees C isothermal conditions. The sensitivity of tests for S. suis 2 detection varies between detection methods and reaction systems, indicating that for each LAMP reaction system, multiple detection methods should be performed to select the optimal one. The sensitivities of the optimized methods (7.16 copies/reaction) in the present study were identical to those of the real-time PCR assay, and the test results for reference strains and clinical samples showed that these LAMP systems have high specificities. Thus, since the LAMP systems established in this study are simple, fast, and sensitive, they may have good clinical potential for detecting the highly pathogenic S. suis 2.