Sethabutr O, Echeverria P, Hoge C W, Bodhidatta L, Pitarangsi C
Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand.
J Diarrhoeal Dis Res. 1994 Dec;12(4):265-9.
The rate of detection of Shigella and enteroinvasive Escherichia coli (EIEC) using a PCR technique was compared with the rate detected by standard microbiological methods (bacteriology plus hybridization of E. coli colonies with a 17 kb EIEC probe) among patients with dysentery before and after antibiotic therapy. The PCR amplified DNA sequences encoding IpaH, a multiple copy sequence located on the chromosome and the invasion plasmid. Shigella or EIEC were detected using the IpaH PCR system among 72 (61%) of 119 patients with dysentery on the first day they were seen at hospital, compared to 50 (42%) using standard microbiological methods (p = 0.006). After three days of antibiotic therapy, IpaH sequences were detected in stools from 38 percent of patients, compared to 10 percent using standard microbiology (p < 0.001). After seven days of therapy, the rates were 26 percent vs. 8 percent respectively (p < 0.001). The IpaH PCR system appeared to be specific for Shigella or EIEC based on low rates of positive reactions among non-diarrhoea controls, and a strong correlation between persistently positive reactions and antibiotic resistance of bacterial isolates. IpaH sequences were detected in 10 (8%) of 119 drinking water samples from homes of patients with disease; none of these specimens were positive for Shigella or EIEC by standard microbiology. In conclusion, PCR amplification of IpaH sequences and detection of target DNA with a non-radioactive probe increased the rates of identification of Shigella and EIEC by 45% in initial clinical specimens and by nearly 300% in specimens obtained from patients receiving antibiotic therapy.
采用聚合酶链反应(PCR)技术检测志贺氏菌和肠侵袭性大肠杆菌(EIEC)的检出率,并与标准微生物学方法(细菌学检测加用17kb EIEC探针与大肠杆菌菌落杂交)在抗生素治疗前后的痢疾患者中的检出率进行比较。PCR扩增编码IpaH的DNA序列,IpaH是位于染色体和侵袭质粒上的多拷贝序列。在119例痢疾患者入院第一天,使用IpaH PCR系统检测到72例(61%)志贺氏菌或EIEC,而使用标准微生物学方法检测到50例(42%)(p = 0.006)。抗生素治疗三天后,38%的患者粪便中检测到IpaH序列,而使用标准微生物学方法检测到的比例为10%(p < 0.001)。治疗七天后,这一比例分别为26%和8%(p < 0.001)。基于非腹泻对照中阳性反应率较低以及持续阳性反应与细菌分离株抗生素耐药性之间的强相关性,IpaH PCR系统似乎对志贺氏菌或EIEC具有特异性。在119份来自患病患者家庭的饮用水样本中,有10份(8%)检测到IpaH序列;通过标准微生物学方法,这些样本中均未检测到志贺氏菌或EIEC阳性。总之,IpaH序列的PCR扩增和使用非放射性探针检测靶DNA,使初始临床标本中志贺氏菌和EIEC的鉴定率提高了45%,在接受抗生素治疗患者的标本中提高了近300%。