Caeiro J P, Estrada-Garcia M T, Jiang Z D, Mathewson J J, Adachi J A, Steffen R, DuPont H L
Department of Medicine, Baylor College of Medicine, Houston, Texas, USA.
J Infect Dis. 1999 Dec;180(6):2053-5. doi: 10.1086/315121.
This study sought to determine whether a specific polymerase chain reaction (PCR) for enterotoxigenic Escherichia coli (ETEC) toxins after chaotropic extraction of DNA from stool would increase the detection of ETEC over that of conventional oligonucleotide probe hybridization of 5 E. coli colonies per stool sample (a standard method). By DNA hybridization, 29 (21%) of 140 patients were positive for ETEC, and 59 (42%) of 140 were positive for ETEC when PCR was used. Sensitivity of the PCR assay was confirmed through spiked stool experiments to be approximately 100-1000 ETEC colonies per sample. Specificity of the assay was determined by showing an absence of ETEC by the PCR technique in a subgroup of 48 subjects and by confirming the presence of ETEC DNA of positive samples by dot blot procedure. PCR technique detected significantly more ETEC infections in these subjects than did the hybridization method (P<.0001).
本研究旨在确定,在对粪便中的DNA进行离液剂提取后,针对产肠毒素大肠杆菌(ETEC)毒素的特异性聚合酶链反应(PCR),与传统的每个粪便样本对5个大肠杆菌菌落进行寡核苷酸探针杂交法(一种标准方法)相比,是否能提高ETEC的检测率。通过DNA杂交,140名患者中有29名(21%)ETEC呈阳性;使用PCR时,140名患者中有59名(42%)ETEC呈阳性。通过加标粪便实验证实,PCR检测的灵敏度约为每个样本100 - 1000个ETEC菌落。通过在48名受试者的亚组中用PCR技术显示未检测到ETEC,并通过斑点印迹法确认阳性样本中ETEC DNA的存在,来确定该检测方法的特异性。在这些受试者中,PCR技术检测到的ETEC感染明显多于杂交法(P <.0001)。