Groseth A, Jones S, Artsob H, Feldmann H, 2005. Hemorrhagic fever viruses as biological weapons. Fong I, Alibek K, eds. Bioterrorism and Infectious Agents: A New Dilemma for the 21st Century. New York: Springer Publishing, 169–191.
Borio L, Inglesby T, Peters CJ, Schmaljohn AL, Hughes JM, Jahrling PB, Ksiazek T, Johnson KM, Meyerhoff A, O'Toole T, Ascher MS, Bartlett J, Breman JG, Eitzen EM Jr., Hamburg M, Hauer J, Henderson DA, Johnson RT, Kwik G, Layton M, Lillibridge S, Nabel GJ, Osterholm MT, Perl TM, Russell P, Tonat K for the Working Group on Civilian Biodefense, 2002. Hemorrhagic fever viruses as biological weapons: medical and public health management. JAMA 287: 2391–2405.
Gibb TR, Norwood DA Jr, Woollen N, Henchal EA, 2001. Development and evaluation of a fluorogenic 5′ nuclease assay to detect and differentiate between Ebola virus subtypes Zaire and Sudan. J Clin Microbiol 39: 4125–4130.
Weidmann M, Muhlberger E, Hufert FT, 2004. Rapid detection protocol for filoviruses. J Clin Virol 30: 94–99.
Sanchez A, Lukwiya M, Bausch D, Mahanty S, Sanchez AJ, Wagoner KD, Rollin PE, 2004. Analysis of human peripheral blood samples from fatal and nonfatal cases of Ebola (Sudan) hemorrhagic fever: cellular responses, virus load, and nitric oxide levels. J Virol 78: 10370–10377.
Towner JS, Sealy TK, Khristova ML, Albarino CG, Conlan S, Reeder SA, Quan PL, Lipkin WI, Downing R, Tappero JW, Okware S, Lutwama J, Bakamutumaho B, Kayiwa J, Comer JA, Rollin PE, Ksiazek TG, Nichol ST, 2008. Newly discovered ebola virus associated with hemorrhagic fever outbreak in Uganda. PLoS Pathog 4: e1000212.
Towner JS, Rollin PE, Bausch DG, Sanchez A, Crary SM, Vincent M, Lee WF, Spiropoulou CF, Ksiazek TG, Lukwiya M, Kaducu F, Downing R, Nichol ST, 2004. 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. J Virol 78: 4330–4341.
Drosten C, Gottig S, Schilling S, Asper M, Panning M, Schmitz H, Gunther S, 2002. Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean–Congo hemorrhagic fever virus, Rift valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR. J Clin Microbiol 40: 2323–2330.
Gibb TR, Norwood DA Jr, Woollen N, Henchal EA, 2001. Development and evaluation of a fluorogenic 5′-nuclease assay to identify Marburg virus. Mol Cell Probes 15: 259–266.
Trappier SG, Conaty AL, Farrar BB, Auperin DD, McCormick JB, Fisher-Hoch SP, 1993. Evaluation of the polymerase chain reaction for diagnosis of Lassa virus infection. Am J Trop Med Hyg 49: 214–221.
Demby AH, Chamberlain J, Brown DW, Clegg CS, 1994. Early diagnosis of Lassa fever by reverse transcription-PCR. J Clin Microbiol 32: 2898–2903.
Lunkenheimer K, Hufert FT, Schmitz H, 1990. Detection of Lassa virus RNA in specimens from patients with Lassa fever by using the polymerase chain reaction. J Clin Microbiol 28: 2689–2692.
Lozano ME, Enria D, Maiztegui JI, Grau O, Romanowski V, 1995. Rapid diagnosis of Argentine hemorrhagic fever by reverse transcriptase PCR-based assay. J Clin Microbiol 33: 1327–1332.
Garin D, Peyrefitte C, Crance JM, Le FA, Jouan A, Bouloy M, 2001. Highly sensitive Taqman PCR detection of Puumala hantavirus. Microbes Infect 3: 739–745.
Weidmann M, Rudaz V, Nunes MR, Vasconcelos PF, Hufert FT, 2003. Rapid detection of human pathogenic orthobunyaviruses. J Clin Microbiol 41: 3299–3305.
Botten J, Mirowsky K, Kusewitt D, Ye C, Gottlieb K, Prescott J, Hjelle B, 2003. Persistent Sin Nombre virus infection in the deer mouse (Peromyscus maniculatus) model: sites of replication and strand-specific expression. J Virol 77: 1540–1550.
Evander M, Eriksson I, Pettersson L, Juto P, Ahlm C, Olsson GE, Bucht G, Allard A, 2007. Puumala hantavirus viremia diagnosed by real-time reverse transcriptase PCR using samples from patients with hemorrhagic fever and renal syndrome. J Clin Microbiol 45: 2491–2497.
Aitichou M, Saleh SS, McElroy AK, Schmaljohn C, Ibrahim MS, 2005. Identification of Dobrava, Hantaan, Seoul, and Puumala viruses by one-step real-time RT-PCR. J Virol Methods 124: 21–26.
Coyne SR, Trombley A, Craw PD, Kulesh DA, Norwood DA, 2008. Extraction of RNA from virus samples in TRIzol using manual and automated magnetic bead systems. American Society for Microbiology Biodefense and Emerging Diseases Research Meeting. Abstract 178 (B): 60–61.
Christensen DR, Hartman LJ, Loveless BM, Frye MS, Shipley MA, Bridge DL, Richards MJ, Kaplan RS, Garrison J, Baldwin CD, Kulesh DA, Norwood DA, 2006. Detection of biological threat agents by real-time PCR: comparison of assay performance on the R.A.P.I.D., the LightCycler, and the Smart Cycler platforms. Clin Chem 52: 141–145.
McCormick JB, Webb PA, Krebs JW, Johnson KM, Smith ES, 1987. A prospective study of the epidemiology and ecology of Lassa fever. J Infect Dis 155: 437–444.
Khan SH, Goba A, Chu M, Roth C, Healing T, Marx A, Fair J, Guttieri MC, Ferro P, Imes T, Monagin C, Garry RF, Bausch DG, 2008. New opportunities for field research on the pathogenesis and treatment of Lassa fever. Antiviral Res 78: 103–115.
Bausch DG, Demby AH, Coulibaly M, Kanu J, Goba A, Bah A, Conde N, Wurtzel HL, Cavallaro KF, Lloyd E, Baldet FB, Cisse SD, Fofona D, Savane IK, Tolno RT, Mahy B, Wagoner KD, Ksiazek TG, Peters CJ, Rollin PE, 2001. Lassa fever in Guinea: I. Epidemiology of human disease and clinical observations. Vector Borne Zoonotic Dis 1: 269–281.
Bausch DG, Sesay SS, Oshin B, 2004. On the front lines of Lassa fever. Emerg Infect Dis 10: 1889–1890.
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Viral hemorrhagic fever is caused by a diverse group of single-stranded, negative-sense or positive-sense RNA viruses belonging to the families Filoviridae (Ebola and Marburg), Arenaviridae (Lassa, Junin, Machupo, Sabia, and Guanarito), and Bunyaviridae (hantavirus). Disease characteristics in these families mark each with the potential to be used as a biological threat agent. Because other diseases have similar clinical symptoms, specific laboratory diagnostic tests are necessary to provide the differential diagnosis during outbreaks and for instituting acceptable quarantine procedures. We designed 48 TaqMan™-based polymerase chain reaction (PCR) assays for specific and absolute quantitative detection of multiple hemorrhagic fever viruses. Forty-six assays were determined to be virus-specific, and two were designated as pan assays for Marburg virus. The limit of detection for the assays ranged from 10 to 0.001 plaque-forming units (PFU)/PCR. Although these real-time hemorrhagic fever virus assays are qualitative (presence of target), they are also quantitative (measure a single DNA/RNA target sequence in an unknown sample and express the final results as an absolute value (e.g., viral load, PFUs, or copies/mL) on the basis of concentration of standard samples and can be used in viral load, vaccine, and antiviral drug studies.
Financial support: This study was supported by the Defense Threat Reduction Agency, project ID no. 8.10007_05_RD_B.
Authors' addresses: Adrienne R. Trombley, Randal J. Schoepp, David A. Norwood, and David A. Kulesh, Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD. Leslie Wachter, Medimmune, Gaithersburg, MD. Jeff Garrison, Battelle, Columbus, OH. Valerie A. Buckley-Beason and Lisa E. Hensley, Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD. Jordan Jahrling, Department of Neuroscience, University of Texas Medical Branch, Galveston, TX. Augustine Goba, Lassa Fever Laboratory, Kenema Government Hospital, Kenema, Sierra Leone. Joseph N. Fair, The Global Viral Forecasting Initiative, San Francisco, CA.
Groseth A, Jones S, Artsob H, Feldmann H, 2005. Hemorrhagic fever viruses as biological weapons. Fong I, Alibek K, eds. Bioterrorism and Infectious Agents: A New Dilemma for the 21st Century. New York: Springer Publishing, 169–191.
Borio L, Inglesby T, Peters CJ, Schmaljohn AL, Hughes JM, Jahrling PB, Ksiazek T, Johnson KM, Meyerhoff A, O'Toole T, Ascher MS, Bartlett J, Breman JG, Eitzen EM Jr., Hamburg M, Hauer J, Henderson DA, Johnson RT, Kwik G, Layton M, Lillibridge S, Nabel GJ, Osterholm MT, Perl TM, Russell P, Tonat K for the Working Group on Civilian Biodefense, 2002. Hemorrhagic fever viruses as biological weapons: medical and public health management. JAMA 287: 2391–2405.
Gibb TR, Norwood DA Jr, Woollen N, Henchal EA, 2001. Development and evaluation of a fluorogenic 5′ nuclease assay to detect and differentiate between Ebola virus subtypes Zaire and Sudan. J Clin Microbiol 39: 4125–4130.
Weidmann M, Muhlberger E, Hufert FT, 2004. Rapid detection protocol for filoviruses. J Clin Virol 30: 94–99.
Sanchez A, Lukwiya M, Bausch D, Mahanty S, Sanchez AJ, Wagoner KD, Rollin PE, 2004. Analysis of human peripheral blood samples from fatal and nonfatal cases of Ebola (Sudan) hemorrhagic fever: cellular responses, virus load, and nitric oxide levels. J Virol 78: 10370–10377.
Towner JS, Sealy TK, Khristova ML, Albarino CG, Conlan S, Reeder SA, Quan PL, Lipkin WI, Downing R, Tappero JW, Okware S, Lutwama J, Bakamutumaho B, Kayiwa J, Comer JA, Rollin PE, Ksiazek TG, Nichol ST, 2008. Newly discovered ebola virus associated with hemorrhagic fever outbreak in Uganda. PLoS Pathog 4: e1000212.
Towner JS, Rollin PE, Bausch DG, Sanchez A, Crary SM, Vincent M, Lee WF, Spiropoulou CF, Ksiazek TG, Lukwiya M, Kaducu F, Downing R, Nichol ST, 2004. 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. J Virol 78: 4330–4341.
Drosten C, Gottig S, Schilling S, Asper M, Panning M, Schmitz H, Gunther S, 2002. Rapid detection and quantification of RNA of Ebola and Marburg viruses, Lassa virus, Crimean–Congo hemorrhagic fever virus, Rift valley fever virus, dengue virus, and yellow fever virus by real-time reverse transcription-PCR. J Clin Microbiol 40: 2323–2330.
Gibb TR, Norwood DA Jr, Woollen N, Henchal EA, 2001. Development and evaluation of a fluorogenic 5′-nuclease assay to identify Marburg virus. Mol Cell Probes 15: 259–266.
Trappier SG, Conaty AL, Farrar BB, Auperin DD, McCormick JB, Fisher-Hoch SP, 1993. Evaluation of the polymerase chain reaction for diagnosis of Lassa virus infection. Am J Trop Med Hyg 49: 214–221.
Demby AH, Chamberlain J, Brown DW, Clegg CS, 1994. Early diagnosis of Lassa fever by reverse transcription-PCR. J Clin Microbiol 32: 2898–2903.
Lunkenheimer K, Hufert FT, Schmitz H, 1990. Detection of Lassa virus RNA in specimens from patients with Lassa fever by using the polymerase chain reaction. J Clin Microbiol 28: 2689–2692.
Lozano ME, Enria D, Maiztegui JI, Grau O, Romanowski V, 1995. Rapid diagnosis of Argentine hemorrhagic fever by reverse transcriptase PCR-based assay. J Clin Microbiol 33: 1327–1332.
Garin D, Peyrefitte C, Crance JM, Le FA, Jouan A, Bouloy M, 2001. Highly sensitive Taqman PCR detection of Puumala hantavirus. Microbes Infect 3: 739–745.
Weidmann M, Rudaz V, Nunes MR, Vasconcelos PF, Hufert FT, 2003. Rapid detection of human pathogenic orthobunyaviruses. J Clin Microbiol 41: 3299–3305.
Botten J, Mirowsky K, Kusewitt D, Ye C, Gottlieb K, Prescott J, Hjelle B, 2003. Persistent Sin Nombre virus infection in the deer mouse (Peromyscus maniculatus) model: sites of replication and strand-specific expression. J Virol 77: 1540–1550.
Evander M, Eriksson I, Pettersson L, Juto P, Ahlm C, Olsson GE, Bucht G, Allard A, 2007. Puumala hantavirus viremia diagnosed by real-time reverse transcriptase PCR using samples from patients with hemorrhagic fever and renal syndrome. J Clin Microbiol 45: 2491–2497.
Aitichou M, Saleh SS, McElroy AK, Schmaljohn C, Ibrahim MS, 2005. Identification of Dobrava, Hantaan, Seoul, and Puumala viruses by one-step real-time RT-PCR. J Virol Methods 124: 21–26.
Coyne SR, Trombley A, Craw PD, Kulesh DA, Norwood DA, 2008. Extraction of RNA from virus samples in TRIzol using manual and automated magnetic bead systems. American Society for Microbiology Biodefense and Emerging Diseases Research Meeting. Abstract 178 (B): 60–61.
Christensen DR, Hartman LJ, Loveless BM, Frye MS, Shipley MA, Bridge DL, Richards MJ, Kaplan RS, Garrison J, Baldwin CD, Kulesh DA, Norwood DA, 2006. Detection of biological threat agents by real-time PCR: comparison of assay performance on the R.A.P.I.D., the LightCycler, and the Smart Cycler platforms. Clin Chem 52: 141–145.
McCormick JB, Webb PA, Krebs JW, Johnson KM, Smith ES, 1987. A prospective study of the epidemiology and ecology of Lassa fever. J Infect Dis 155: 437–444.
Khan SH, Goba A, Chu M, Roth C, Healing T, Marx A, Fair J, Guttieri MC, Ferro P, Imes T, Monagin C, Garry RF, Bausch DG, 2008. New opportunities for field research on the pathogenesis and treatment of Lassa fever. Antiviral Res 78: 103–115.
Bausch DG, Demby AH, Coulibaly M, Kanu J, Goba A, Bah A, Conde N, Wurtzel HL, Cavallaro KF, Lloyd E, Baldet FB, Cisse SD, Fofona D, Savane IK, Tolno RT, Mahy B, Wagoner KD, Ksiazek TG, Peters CJ, Rollin PE, 2001. Lassa fever in Guinea: I. Epidemiology of human disease and clinical observations. Vector Borne Zoonotic Dis 1: 269–281.
Bausch DG, Sesay SS, Oshin B, 2004. On the front lines of Lassa fever. Emerg Infect Dis 10: 1889–1890.
Past two years | Past Year | Past 30 Days | |
---|---|---|---|
Abstract Views | 3422 | 2747 | 120 |
Full Text Views | 1067 | 54 | 22 |
PDF Downloads | 617 | 51 | 5 |