Development of a TaqMan Array Card for Acute-Febrile-Illness Outbreak Investigation and Surveillance of Emerging Pathogens, Including Ebola Virus.
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.
Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, USA.
Jie
J
Liu
Center for Global Health Research, Kenya Medical Research Institute, Nairobi, Kenya.
Caroline
C
Ochieng
Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Dar es Salaam, Tanzania.
Steve
S
Wiersma
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of High Consequence Pathogens and Pathology, Atlanta, Georgia, USA.
Ute
U
Ströher
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of High Consequence Pathogens and Pathology, Atlanta, Georgia, USA.
Jonathan S
JS
Towner
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of High Consequence Pathogens and Pathology, Atlanta, Georgia, USA.
Shannon
S
Whitmer
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of High Consequence Pathogens and Pathology, Atlanta, Georgia, USA.
Stuart T
ST
Nichol
Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, USA.
Christopher C
CC
Moore
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of Vector Borne Diseases, Atlanta, Georgia, USA.
Gilbert J
GJ
Kersh
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of Vector Borne Diseases, Atlanta, Georgia, USA.
Cecilia
C
Kato
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of Vector Borne Diseases, Atlanta, Georgia, USA.
Christopher
C
Sexton
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of Vector Borne Diseases, Atlanta, Georgia, USA.
Jeannine
J
Petersen
Centers for Disease Control and Prevention, National Center for Emerging and Zoonotic Infectious Diseases, Division of High Consequence Pathogens and Pathology, Atlanta, Georgia, USA.
Robert
R
Massung
Georgetown University, Washington, DC, USA.
Christine
C
Hercik
Division of Infectious Diseases and International Health, Duke University, Durham, North Carolina, USA Centre for International Health, University of Otago, Dunedin, New Zealand.
John A
JA
Crump
Kilimanjaro Clinical Research Institute, Moshi, Tanzania.
Gibson
G
Kibiki
Kilimanjaro Clinical Research Institute, Moshi, Tanzania.
Athanasia
A
Maro
Kilimanjaro Clinical Research Institute, Moshi, Tanzania.
Buliga
B
Mujaga
Kilimanjaro Clinical Research Institute, Moshi, Tanzania.
Jean
J
Gratz
Division of Allergy and Infectious Diseases, University of Washington, Seattle, Washington, USA.
Shevin T
ST
Jacob
Masaka Regional Referral Hospital, Masaka, Uganda.
Patrick
P
Banura
Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, USA.
W Michael
WM
Scheld
Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Nairobi, Kenya.
Bonventure
B
Juma
Centers for Disease Control and Prevention, Center for Global Health, Division of Global Health Protection, Nairobi, Kenya.
Clayton O
CO
Onyango
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.
Joel M
JM
Montgomery
Division of Infectious Diseases and International Health, University of Virginia, Charlottesville, Virginia, USA erh6k@virginia.edu bsf2@cdc.gov.
Eric
E
Houpt
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.
Barry
B
Fields
D000328:Adult; D003141:Communicable Diseases / Q000175:diagnosis* / Q000453:epidemiology*; D004196:Disease Outbreaks; D062665:Epidemiological Monitoring; D005335:Fever of Unknown Origin / Q000175:diagnosis* / Q000453:epidemiology*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000592:standards; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity; D013997:Time Factors