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标题：Nanolitre real-time PCR detection of bacterial, parasitic, and viral agents from patients with diarrhoea in Nunavut, Canada
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作者：David M. Goldfarb ; Brent Dixon ; Ioana Moldovan 等
期刊名称：International Journal of Circumpolar Health
印刷版ISSN：1239-9736
电子版ISSN：2242-3982
出版年度：2013
卷号：72
DOI：10.3402/ijch.v72i0.19903
语种：English
摘要：Background. Little is known about the microbiology of diarrhoeal disease in Canada's Arctic regions. There are a number of limitations of conventional microbiology testing techniques for diarrhoeal pathogens, and these may be further compromised in the Arctic, given the often long distances for specimen transport. Objective. To develop a novel multiple-target nanolitre real-time reverse transcriptase (RT)-PCR platform to simultaneously test diarrhoeal specimens collected from residents of the Qikiqtani (Baffin Island) Region of Nunavut, Canada, for a wide range of bacterial, parasitic and viral agents. Study design/methods. Diarrhoeal stool samples submitted for bacterial culture to Qikiqtani General Hospital in Nunavut over an 18-month period were tested with a multiple-target nanolitre real-time PCR panel for major diarrhoeal pathogens including 8 bacterial, 6 viral and 2 parasitic targets. Results. Among 86 stool specimens tested by PCR, a total of 50 pathogens were detected with 1 or more pathogens found in 40 (46.5%) stool specimens. The organisms detected comprised 17 Cryptosporidium spp., 5 Clostridium difficile with toxin B, 6 Campylobacter spp., 6 Salmonella spp., 4 astroviruses, 3 noroviruses, 1 rotavirus, 1 Shigella spp. and 1 Giardia spp. The frequency of detection by PCR and bacterial culture was similar for Salmonella spp., but discrepant for Campylobacter spp., as Campylobacter was detected by culture from only 1/86 specimens. Similarly, Cryptosporidium spp. was detected in multiple samples by PCR but was not detected by microscopy or enzyme immunoassay. Conclusions. Cryptosporidium spp., Campylobacter spp. and Clostridium difficile may be relatively common but possibly under-recognised pathogens in this region. Further study is needed to determine the regional epidemiology and clinical significance of these organisms. This method appears to be a useful tool for gastrointestinal pathogen research and may also be helpful for clinical diagnostics and outbreak investigation in remote regions where the yield of routine testing may be compromised.Keywords: multiplex real-time PCR; detection; pathogens; Campylobacter spp.; Cryptosporidium spp.; Clostridium difficile; viral agents; diarrhoea; gastroenteritis; Nunavut; Arctic(Published: 8 April 2013)Citation: Int J Circumpolar Health 2013, 72: 19903 - http://dx.doi.org/10.3402/ijch.v72i0.19903
关键词：Microbiology; Infectious diseases; Gastroenterology;Multiplex Real-Time PCR; Detection; Pathogens, Campylobacter spp.; Cryptosporidium spp.; Clostridium difficile; Viral Agents; Diarrhea; Gastroenteritis; Nunavut; Arctic