Division of Clinical Microbiology, Department of Laboratory Medicine and Pathology, Mayo Clinic, 200 First St S.W., Rochester, MN 55905, USA.
J Clin Microbiol. 2010 Aug;48(8):2929-33. doi: 10.1128/JCM.00339-10. Epub 2010 Jun 2.
Campylobacter jejuni and Salmonella, Shigella, and Yersinia species (along with Shiga toxin-producing Escherichia coli) are the most common causes of acute bacterial diarrheal disease in the United States. Current detection techniques are time-consuming, limiting usefulness for patient care. We developed and validated a panel of rapid PCR assays for the detection and identification of C. jejuni, C. coli, Salmonella, and Yersinia species and Shigella and enteroinvasive E. coli in stool samples. A total of 392 archived stool specimens, previously cultured for enteric pathogens, were evaluated by PCR. Overall, 104 stool specimens had been culture positive (C. jejuni/coli [n = 51], Salmonella species [n = 42], Shigella species [n = 6], and Yersinia species [n = 5]). Compared to culture, the overall sensitivity and specificity of PCR detection of these organisms were 92 and 98% (96/104 and 283/288), respectively, from fresh or Cary Blair stool (P = 0.41); 87 and 98% (41/47 and 242/246), respectively, from fresh stool (P = 0.53); and 96 and 98% (55/57 and 41/42), respectively, from Cary Blair stool (P = 0.56). For individual genera, PCR was as sensitive as the culture method, with the exception of Salmonella culture using selenite enrichment for which PCR was less sensitive than culture from fresh, but not Cary Blair (P = 0.03 and 1.00, respectively) stools. This PCR assay panel for the rapid diagnosis of acute infectious bacterial diarrheal pathogens has a sensitivity and specificity equivalent to that of culture for stools in Cary Blair transport medium. Paired with reflexive culture of stools testing positive, this should provide an improvement in care for patients with acute infectious diarrheal disease.
空肠弯曲菌和沙门氏菌、志贺氏菌和耶尔森氏菌(以及产志贺毒素的大肠杆菌)是美国急性细菌性腹泻病最常见的病因。目前的检测技术耗时较长,限制了其在患者护理中的应用。我们开发并验证了一组快速 PCR 检测方法,用于检测和鉴定粪便样本中的空肠弯曲菌、大肠弯曲菌、沙门氏菌、耶尔森氏菌和志贺氏菌以及侵袭性大肠杆菌。总共评估了 392 份存档的粪便标本,这些标本之前曾培养过肠道病原体。总的来说,104 份粪便标本培养呈阳性(空肠弯曲菌/大肠弯曲菌[n = 51]、沙门氏菌[n = 42]、志贺氏菌[n = 6]和耶尔森氏菌[n = 5])。与培养相比,这些病原体的 PCR 检测总体敏感性和特异性分别为 92%和 98%(96/104 和 283/288),新鲜或 Cary Blair 粪便(P = 0.41);新鲜粪便时分别为 87%和 98%(41/47 和 242/246)(P = 0.53);Cary Blair 粪便时分别为 96%和 98%(55/57 和 41/42)(P = 0.56)。对于个别属,PCR 的敏感性与培养方法相当,除了使用亚硒酸盐富集法培养的沙门氏菌,PCR 检测从新鲜但不是 Cary Blair 粪便(P = 0.03 和 1.00)中检测沙门氏菌的敏感性低于培养法。该快速诊断急性感染性细菌性腹泻病原体的 PCR 检测方法在 Cary Blair 运输培养基中的粪便检测中具有与培养法相当的敏感性和特异性。与对检测呈阳性的粪便进行反射性培养相结合,这将为急性感染性腹泻病患者的治疗提供改善。