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J Clin Microbiol. 2016 Jan;54(1):49-58. doi: 10.1128/JCM.02257-15. Epub 2015 Oct 21.

Development of a TaqMan Array Card for Acute-Febrile-Illness Outbreak Investigation and Surveillance of Emerging Pathogens, Including Ebola Virus.

Author information

1
Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, USA.
2
Center for Global Health Research, Kenya Medical Research Institute, Nairobi, Kenya.
3
Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Dar es Salaam, Tanzania.
4
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of High Consequence Pathogens and Pathology, Atlanta, Georgia, USA.
5
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of Vector Borne Diseases, Atlanta, Georgia, USA.
6
Georgetown University, Washington, DC, USA.
7
Division of Infectious Diseases and International Health, Duke University, Durham, North Carolina, USA Centre for International Health, University of Otago, Dunedin, New Zealand.
8
Kilimanjaro Clinical Research Institute, Moshi, Tanzania.
9
Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, USA.
10
Masaka Regional Referral Hospital, Masaka, Uganda.
11
Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Nairobi, Kenya.
12
Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Nairobi, Kenya Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Atlanta, Georgia, USA.
13
Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, USA erh6k@virginia.edu bsf2@cdc.gov.
14
Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Nairobi, Kenya Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Atlanta, Georgia, USA erh6k@virginia.edu bsf2@cdc.gov.

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.

PMID:
26491176
PMCID:
PMC4702733
DOI:
10.1128/JCM.02257-15
[Indexed for MEDLINE]
Free PMC Article

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