Rundell Mark S, Pingle Maneesh, Das Sanchita, Hussain Aashiq, Ocheretina Oksana, Charles Macarthur, Larone Davise H, Spitzer Eric D, Golightly Linnie, Barany Francis
Department of Microbiology and Immunology, Weill Medical College of Cornell University, Box 62, New York, NY 10021.
Department of Medicine, Division of International Medicine and Infectious Diseases, Weill Medical College of Cornell University, Box 62, New York, NY 10021.
Diagn Microbiol Infect Dis. 2014 Jun;79(2):135-40. doi: 10.1016/j.diagmicrobio.2014.02.022. Epub 2014 Mar 12.
Enteric pathogens that cause gastroenteritis remain a major global health concern. The goal of this study was to develop a multiplex PCR/ligation detection reaction (LDR) assay for the detection of all NIAID category B bacterial food and water-borne pathogens directly from stool specimens. To validate the PCR/LDR assay, clinical isolates of Campylobacter spp., Vibrio spp., Shigella spp., Salmonella spp., Listeria monocytogenes, Yersinia enterocolitica, and diarrheagenic Escherichia coli were tested. The sensitivity and specificity of the assay were assessed using a large number of seeded culture-negative stool specimens and a smaller set of clinical specimens from Haiti. The overall sensitivity ranged from 91% to 100% (median 100%) depending on the species. For the majority of organisms, the sensitivity was 100%. The overall specificity based on initial testing ranged from 98% to 100% depending on the species. After additional testing of discordant samples, the lowest specificity was 99.4%. PCR/LDR detected additional category B agents (particularly diarrheagenic E. coli) in 11/40 specimens from Haiti that were culture-positive for V. cholerae and in approximately 1% of routine culture-negative stool specimens from a hospital in New York. This study demonstrated the ability of the PCR/LDR assay to detect a large comprehensive panel of category B enteric bacterial pathogens as well as mixed infections. This type of assay has the potential to provide earlier warnings of possible public health threats and more accurate surveillance of food and water-borne pathogens.
引起肠胃炎的肠道病原体仍是全球主要的健康问题。本研究的目的是开发一种多重聚合酶链反应/连接检测反应（LDR）检测方法，用于直接从粪便标本中检测美国国立过敏与传染病研究所（NIAID）B类所有食源和水源性细菌病原体。为验证聚合酶链反应/LDR检测方法，对弯曲杆菌属、弧菌属、志贺菌属、沙门菌属、单核细胞增生李斯特菌、小肠结肠炎耶尔森菌和致泻性大肠杆菌的临床分离株进行了检测。使用大量接种培养阴性的粪便标本和来自海地的少量临床标本评估了该检测方法的敏感性和特异性。总体敏感性根据菌种不同在91%至100%之间（中位数为100%）。对于大多数生物体，敏感性为100%。基于初始检测的总体特异性根据菌种不同在98%至100%之间。对不一致样本进行额外检测后，最低特异性为99.4%。聚合酶链反应/LDR在来自海地的40份霍乱弧菌培养阳性标本中的11份以及纽约一家医院约1%的常规培养阴性粪便标本中检测到了其他B类病原体（特别是致泻性大肠杆菌）。本研究证明了聚合酶链反应/LDR检测方法能够检测一大组综合性的B类肠道细菌病原体以及混合感染。这种检测方法有可能对可能的公共卫生威胁提供早期预警，并对食源和水源性病原体进行更准确的监测。
