猪链球菌2型荧光PCR检测方法与常规检测方法的比较

Comparative Detection of Streptococcus suis Serotype 2 by Real-time PCR and conventional detection methods

关键词:  猪链球菌2型  检测方法  比较

Key words:  Streptococcus suis serotypeⅡ  test method  comparison

摘要:

荧光PCR检测技术是90年代末推出的一种分子生物学检测新技术。为了验证猪链球菌2型荧光PCR检测方法对临床样品检测的敏感性和适用性，以及验证该方法所拥有的独特优点，本项研究用荧光PCR法、常规PCR法和细菌分离法对人工感染猪链球菌2型的小鼠肝脏和发病猪的心、肝、、脾、肾等实质器官和血液、喉拭子进行抗原检测。结果荧光PCR法检出率为70.8%，明显高于普通PCR法（检出率为20.8%），也高于常规细菌分离法（检出率为42.8%）。由于临床样品常常会被其他细菌污染，细菌分离法很难准确分离到链球菌，该方法不受其他细菌污染的影响。对实验室培养的猪链球菌2型菌液，该方法检测滴度可达10-6（42-52CFU/0.1ml），而普通PCR方法检测滴度仅为10－4。

Abstract:

Real-time PCR detecting Technique was successfully developed in the end of 1990s. In order to validate the clinical sensitivity and applicability of Streptococcus suis serotype 2 Real-time PCR assay, rat liver of artificial infection, parenchymal organs（e.g. heart, liver, spleen, kidney and so on）as well as blood and throat swabs of swine which were challenged Streptococcus suis serotype Ⅱ was performed by comparison with conventional PCR method and bacterial isolation method. The results indicated that detection rate of Real-time PCR assay was 70.8% which obviously higher than that of conventional PCR method (20.8%) and higher than bacterial isolation method (42.8%). Meanwhile, clinical samples were generally contaminated by other bacterium, and could be hardly isolated specific to streptococcus accurately, whereas this assay was not interfered with bacterial contamination. For liquid cultures of Streptococcus suis serotype Ⅱ in laboratory, detected titre of Real-time PCR was 10-6 /0.1 ml （42-45 CFU/0.1ml）, while that of conventional PCR method was just 10-4.