Development of a TaqMan Array Card for Acute-Febrile-Illness Outbreak Investigation and Surveillance of Emerging Pathogens, Including Ebola Virus.
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2016-01
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Abstract
Acute febrile illness (AFI) is associated with substantial morbidity and mortality worldwide, yet an etiologic agent is often not identified. Convalescent-phase serology is impractical, blood culture is slow, and many pathogens are fastidious or impossible to cultivate. We developed a real-time PCR-based TaqMan array card (TAC) that can test six to eight samples within 2.5 h from sample to results and can simultaneously detect 26 AFI-associated organisms, including 15 viruses (chikungunya, Crimean-Congo hemorrhagic fever [CCHF] virus, dengue, Ebola virus, Bundibugyo virus, Sudan virus, hantaviruses [Hantaan and Seoul], hepatitis E, Marburg, Nipah virus, o'nyong-nyong virus, Rift Valley fever virus, West Nile virus, and yellow fever virus), 8 bacteria (Bartonella spp., Brucella spp., Coxiella burnetii, Leptospira spp., Rickettsia spp., Salmonella enterica and Salmonella enterica serovar Typhi, and Yersinia pestis), and 3 protozoa (Leishmania spp., Plasmodium spp., and Trypanosoma brucei). Two extrinsic controls (phocine herpesvirus 1 and bacteriophage MS2) were included to ensure extraction and amplification efficiency. Analytical validation was performed on spiked specimens for linearity, intra-assay precision, interassay precision, limit of detection, and specificity. The performance of the card on clinical specimens was evaluated with 1,050 blood samples by comparison to the individual real-time PCR assays, and the TAC exhibited an overall 88% (278/315; 95% confidence interval [CI], 84% to 92%) sensitivity and a 99% (5,261/5,326, 98% to 99%) specificity. This TaqMan array card can be used in field settings as a rapid screen for outbreak investigation or for the surveillance of pathogens, including Ebola virus.
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Liu, Jie, Caroline Ochieng, Steve Wiersma, Ute Ströher, Jonathan S Towner, Shannon Whitmer, Stuart T Nichol, Christopher C Moore, et al. (2016). Development of a TaqMan Array Card for Acute-Febrile-Illness Outbreak Investigation and Surveillance of Emerging Pathogens, Including Ebola Virus. J Clin Microbiol, 54(1). pp. 49–58. 10.1128/JCM.02257-15 Retrieved from https://hdl.handle.net/10161/13764.
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John Andrew Crump
I am an Adjunct Professor of Medicine, Pathology, and Global Health. My work with Duke University is primarily based in northern Tanzania where I am former Site Leader and current Principal Investigator on projects linked to Duke University’s collaborative research program at Kilimanjaro Christian Medical Centre. I oversee the design and implementation of research studies on infectious diseases, particularly febrile illness, invasive bacterial disease, zoonotic infections, and infectious diseases diagnostics. In addition, I am Professor of Medicine, Pathology, and Global Health at the University of Otago and a medical epidemiologist with the US Centers for Disease Control and Prevention (CDC). My CDC work focuses on non-malaria febrile illness.
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