PCR检测粪便中志贺菌

Detection of Shigella in Feces by PCR

目的建立一种粪便样本中快速、特异、敏感的志贺菌检测方法。方法运用PCR对痢疾、福氏、鲍氏和宋内4群志贺菌进行侵袭性质粒H抗原(ipaH)基因的检测,并对该PCR方法进行反应的灵敏度、特异性以及模拟粪便样品的最低检出限测定。结果4群志贺菌均能检出侵袭性质粒H抗原(ipaH)基因,纯菌检测灵敏度达102cfu/ml,特异性实验中志贺菌菌株PCR扩增均为阳性,阴性对照菌株未扩增出特异性条带,混合菌株PCR扩增均为阳性;接种志贺菌到粪便样品中,当粪便样本中菌量达104cfu/ml以上时,不需要增菌,志贺菌均可立即检出。结论PCR方法检测志贺菌灵敏度高、快速、特异性强,可以很好地应用于粪便等临床样本中志贺菌的检测与鉴定。

Objective To establish a rapid, sensitive and accurate detection technique for Shigella in the feces. Methods Polymerase chain reaction （PCR） was used to detect ipaH gene from Shigella, The method＇s sentivity, accurate and the detection limit in the artificial feces sample were stndied. Results All Shigella strains could be detected by PCR, the negative control strains couldn＇t be detected, and the mix strains could also be detected. The detection sensitivity was 10^2 cfu/ml in pure culture, while detection limit of artificially contaminated feces sample was 10^4 cfu/ml and didn＇ t need to enrichment. Conclusion Detection of feces sample shows that this method is accurate, sensitive and rapid. PCR assay should be widely used to detect ShigeUa in feces, even in other clinical samples.