Grimes Kevin A, Mohamed Jamal A, Dupont Herbert L, Padda Ranjit S, Jiang Zhi-Dong, Flores Jose, Belkind-Gerson Jaime, Martinez-Sandoval Francisco G, Okhuysen Pablo C
Division of Infectious Diseases, The University of Texas Medical School, Houston, Texas, USA.
J Clin Microbiol. 2008 Jul;46(7):2227-30. doi: 10.1128/JCM.00073-08. Epub 2008 May 14.
Large field studies of travelers' diarrhea for multiple destinations are limited by the need to perform stool cultures on site in a timely manner. A method for the collection, transport, and storage of fecal specimens that does not require immediate processing and refrigeration and that is stable for months would be advantageous. This study was designed to determine if enterotoxigenic Escherichia coli (ETEC) and enteroaggregative E. coli (EAEC) DNA could be identified from cards that were processed for the evaluation of fecal occult blood. U.S. students traveling to Mexico during 2005 to 2007 were monitored for the occurrence of diarrheal illness. When ill, students provided a stool specimen for culture and occult blood by the standard methods. Cards then were stored at room temperature prior to DNA extraction. Fecal PCR was performed to identify ETEC and EAEC in DNA extracted from stools and from occult blood cards. Significantly more EAEC cases were identified by PCR that was performed on DNA that was extracted from cards (49%) or from frozen feces (40%) than from culture methods that used HEp-2 adherence assays (13%) (P < 0.001). Similarly, more ETEC cases were detected from card DNA (38%) than from fecal DNA (30%) or by culture that was followed by hybridization (10%) (P < 0.001). The sensitivity and specificity of the card test were 75 and 62%, respectively, compared to those for EAEC by culture and were 50 and 63%, respectively, compared to those for ETEC. DNA extracted from fecal cards that was used for the detection of occult blood is of use in identifying diarrheagenic E. coli.
针对多个目的地旅行者腹泻的大规模现场研究,因需要及时在现场进行粪便培养而受到限制。一种无需立即处理和冷藏且能稳定保存数月的粪便标本采集、运输及储存方法将很有优势。本研究旨在确定能否从用于粪便潜血评估的卡片中鉴定出产肠毒素大肠杆菌(ETEC)和肠聚集性大肠杆菌(EAEC)的DNA。对2005年至2007年前往墨西哥的美国学生的腹泻病发病情况进行了监测。学生生病时,按标准方法提供粪便标本用于培养和潜血检测。然后在进行DNA提取之前,将卡片室温保存。对从粪便和潜血卡片中提取的DNA进行粪便PCR,以鉴定ETEC和EAEC。通过对从卡片(49%)或冷冻粪便(40%)中提取的DNA进行PCR鉴定出的EAEC病例,显著多于使用HEP-2黏附试验的培养方法(13%)(P<0.001)。同样,从卡片DNA中检测到的ETEC病例(38%)多于从粪便DNA中检测到的(30%)或通过培养后杂交检测到的(10%)(P<0.001)。与通过培养检测EAEC相比,卡片检测的敏感性和特异性分别为75%和62%,与检测ETEC相比,分别为50%和63%。从用于检测潜血的粪便卡片中提取的DNA可用于鉴定致泻性大肠杆菌。