Rapid diagnosis of Ebola hemorrhagic fever by reverse transcription-PCR in an outbreak setting and assessment of patient viral load as a predictor of outcome.
The largest outbreak on record of Ebola hemorrhagic fever (EHF) occurred in Uganda from August 2000 to January 2001. The outbreak was centered in the Gulu district of northern Uganda, with secondary transmission to other districts. After the initial diagnosis of Sudan ebolavirus by the National Institute for Virology in Johannesburg, South Africa, a temporary diagnostic laboratory was established within the Gulu district at St. Mary's Lacor Hospital. The laboratory used antigen capture and reverse transcription-PCR (RT-PCR) to diagnose Sudan ebolavirus infection in suspect patients. The RT-PCR and antigen-capture diagnostic assays proved very effective for detecting ebolavirus in patient serum, plasma, and whole blood. In samples collected very early in the course of infection, the RT-PCR assay could detect ebolavirus 24 to 48 h prior to detection by antigen capture. More than 1,000 blood samples were collected, with multiple samples obtained from many patients throughout the course of infection. Real-time quantitative RT-PCR was used to determine the viral load in multiple samples from patients with fatal and nonfatal cases, and these data were correlated with the disease outcome. RNA copy levels in patients who died averaged 2 log(10) higher than those in patients who survived. Using clinical material from multiple EHF patients, we sequenced the variable region of the glycoprotein. This Sudan ebolavirus strain was not derived from either the earlier Boniface (1976) or Maleo (1979) strain, but it shares a common ancestor with both. Furthermore, both sequence and epidemiologic data are consistent with the outbreak having originated from a single introduction into the human population.
Towner
Jonathan S
JS
Special Pathogens Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
Rollin
Pierre E
PE
Bausch
Daniel G
DG
Sanchez
Anthony
A
Crary
Sharon M
SM
Vincent
Martin
M
Lee
William F
WF
Spiropoulou
Christina F
CF
Ksiazek
Thomas G
TG
Lukwiya
Mathew
M
Kaducu
Felix
F
Downing
Robert
R
Nichol
Stuart T
ST
D000956:Antigens, Viral / Q000097:blood; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D004196:Disease Outbreaks; D029043:Ebolavirus / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis* / Q000453:epidemiology* / Q000821:virology; D006801:Humans; D017720:Molecular Epidemiology; D011379:Prognosis; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D014454:Uganda / Q000453:epidemiology; D014764:Viral Proteins / Q000235:genetics; D014766:Viremia / Q000821:virology
D000956:Antigens, Viral; D004279:DNA, Viral; D012367:RNA, Viral; D014764:Viral Proteins