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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

Rapid Diagnosis of Ebola Hemorrhagic Fever by Reverse Transcription-PCR in an Outbreak Setting... 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 Jonathan S. Towner 1 , Pierre E. Rollin 1 , Daniel G. Bausch 1 , Anthony Sanchez 1 , Sharon M. Crary 1 , 2 , Martin Vincent 1 , William F. Lee 3 , Christina F. Spiropoulou 1 , Thomas G. Ksiazek 1 , Mathew Lukwiya 4 , Felix Kaducu 5 , Robert Downing 6 , and Stuart T. Nichol 1 , * 1 Special Pathogens Branch 3 Department of Pathology, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease ControlPrevention, Atlanta, Georgia 2 Depauw University, Greencastle, Indiana 4 Saint Mary's Lacor Hospital 5 Gulu Regional Hospital, Ministry of Health, Gulu 6 Uganda Virus Research Institute, Centers for Disease Control and Prevention Uganda, Entebbe, Uganda ABSTRACT 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. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Virology American Society For Microbiology

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

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

Journal of Virology , Volume 78 (8): 4330 – Apr 15, 2004

Abstract

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 Jonathan S. Towner 1 , Pierre E. Rollin 1 , Daniel G. Bausch 1 , Anthony Sanchez 1 , Sharon M. Crary 1 , 2 , Martin Vincent 1 , William F. Lee 3 , Christina F. Spiropoulou 1 , Thomas G. Ksiazek 1 , Mathew Lukwiya 4 , Felix Kaducu 5 , Robert Downing 6 , and Stuart T. Nichol 1 , * 1 Special Pathogens Branch 3 Department of Pathology, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease ControlPrevention, Atlanta, Georgia 2 Depauw University, Greencastle, Indiana 4 Saint Mary's Lacor Hospital 5 Gulu Regional Hospital, Ministry of Health, Gulu 6 Uganda Virus Research Institute, Centers for Disease Control and Prevention Uganda, Entebbe, Uganda ABSTRACT 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.

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References (21)

Publisher
American Society For Microbiology
Copyright
Copyright © 2004 by the American society for Microbiology.
ISSN
0022-538X
eISSN
1098-5514
DOI
10.1128/JVI.78.8.4330-4341.2004
Publisher site
See Article on Publisher Site

Abstract

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 Jonathan S. Towner 1 , Pierre E. Rollin 1 , Daniel G. Bausch 1 , Anthony Sanchez 1 , Sharon M. Crary 1 , 2 , Martin Vincent 1 , William F. Lee 3 , Christina F. Spiropoulou 1 , Thomas G. Ksiazek 1 , Mathew Lukwiya 4 , Felix Kaducu 5 , Robert Downing 6 , and Stuart T. Nichol 1 , * 1 Special Pathogens Branch 3 Department of Pathology, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease ControlPrevention, Atlanta, Georgia 2 Depauw University, Greencastle, Indiana 4 Saint Mary's Lacor Hospital 5 Gulu Regional Hospital, Ministry of Health, Gulu 6 Uganda Virus Research Institute, Centers for Disease Control and Prevention Uganda, Entebbe, Uganda ABSTRACT 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.

Journal

Journal of VirologyAmerican Society For Microbiology

Published: Apr 15, 2004

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