自建分子检测方法在腹泻患者艰难梭菌感染调查中的应用

Application of a seLF-developed molecular detection method in investigation of Clostridium difficile infection in patients with diarrhea

目的评估自建的PCR方法对粪便标本中艰难梭菌的直接检测能力。方法收集云南省第一人民医院244例腹泻患者的粪便标本,分别采用PCR方法与传统厌氧培养法对标本进行直接检测,阳性标本进行毒素A/B的PCR直接检测和酶联免疫荧光法检测,利用MLST分型技术对PCR方法检测阳性的粪便标本进行艰难梭菌分型。结果收集的244例腹泻患者粪便标本中艰难梭菌厌氧培养检测出12例阳性,PCR法检测出28例阳性,检出率分别为4.92%和11.48%,差异有统计学意义(P=0.008)。对28例患者阳性粪便标本进行毒素A/B检测显示,PCR法检测出24例阳性,酶联免疫荧光法检出11例阳性,检出率分别为85.71%和39.28%,差异有统计学意义(P=0.0003)。对检出的阳性粪便标本进行MLST分型,结果显示仅存在散发性的患病,没有发生暴发性流行。结论自建的分子诊断方法对粪便标本中艰难梭菌的直接检测能力好,适于实验室对艰难梭菌的快速筛查。

Objective To evaluate the direct detection ability of a self-developed PCR detection method for Clostridium difficile in feces samples.Methods Feces of 244 patients with diarrhea in Yunnan First People′s Hospital were collected.Samples were tested by PCR and traditional anaerobic culture.The positive samples were detected by PCR and enzyme-linked immunofluorescence assay for Toxin A/B.Then MLST typing technology was used to classify Clostridium difficile in positive samples.Results A total of 12 positive clostridium difficile strains were detected in 244 samples by anaerobic culture and 28 were detected by PCR.The detection rate was 4.92%and 11.48%,respectively.The difference was statistically significant(P=0.008).Among the 28 positive samples tested for Toxin A/B,24 were detected by PCR and 11 were detected by enzyme-linked immunofluorescence assay.The detection rate was 85.71%and 39.28%,respectively.The difference was statistically significant(P=0.0003).MLST typing of positive samples showed that only sporadic diseases existed and no outbreaks occurred.Conclusion The self-developed PCR detection method has better direct detection ability for Clostridium difficile in feces samples,which is suitable for rapid and large-scale epidemiological investigation.