- INTRODUCTION
- MATERIALS AND METHODS
- RT-PCR primers, TaqMan®-MGB probe, and target sequences.
- Viral RNA propagation and RNA isolation.
- Real-time RT-PCR assay.
- DNA and RNA panel evaluations.
- Limit of detection.
- Indirect hemagglutination (HI), agarose gel diffusion and precipitation (AGDP), and complement fixation (CF).
- Sequencing the CCHF S segment.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Swanepoel R, Leman PA, Burt FJ, Jardine J, Verwoerd DJ, Capua I, Bruckner GK, Burger WP, 1998. Experimental infection of ostriches with Crimean-Congo haemorrhagic fever virus. Epidemiol Infect 121 :427–432.
-
2
Dohm DJ, Logan TM, Linthicum KJ, Rossi CA, Turell MJ, 1996. Transmission of Crimean-Congo hemorrhagic fever virus by Hyalomma impeltatum (Acari:Ixodidae) after experimental infection. J Med Entomol 33 :848–851.
-
3
Rodriguez LL, Maupin GO, Ksiazek TG, Rollin PE, Khan AS, Schwarz TF, Lofts RS, Smith JF, Noor AM, Peters CJ, Nichol ST, 1997. Molecular investigation of a multisource outbreak of Crimean-Congo hemorrhagic fever in the United Arab Emirates. Am J Trop Med Hyg 57 :512–518.
-
4
Swanepoel R, Shepherd AJ, Leman PA, Shepherd SP, McGillivray GM, Erasmus MJ, Searle LA, Gill DE, 1987. Epidemiologic and clinical features of Crimean-Congo hemorrhagic fever in southern Africa. Am J Trop Med Hyg 36 :120–132.
-
5
Burt FJ, Leman PA, Abbott JC, Swanepoel R, 1994. Serodiagnosis of Crimean-Congo haemorrhagic fever. Epidemiol Infect 113 :551–562.
-
6
Hoogstraal H, 1979. The epidemiology of tick-borne Crimean-Congo hemorrhagic fever in Asia, Europe, and Africa. J Med Entomol 15 :307–417.
-
7
Burney MI, Ghafoor A, Saleen M, Webb PA, Casals J, 1980. Nosocomial outbreak of viral hemorrhagic fever caused by Crimean Hemorrhagic fever-Congo virus in Pakistan, January 1976. Am J Trop Med Hyg 29 :941–947.
-
8
Papa A, Bino S, Llagami A, Brahimaj B, Papadimitriou E, Pavlidou V, Velo E, Cahani G, Hajdini M, Pilaca A, Harxhi A, Antoniadis A, 2002. Crimean-Congo hemorrhagic fever in Albania, 2001. Eur J Clin Microbiol Infect Dis 21 :603–606.
-
9
Shepherd AJ, Swanepoel R, Shepherd SP, Leman PA, Blackburn NK, Hallett AF, 1985. A nosocomial outbreak of Crimean-Congo haemorrhagic fever at Tygerberg Hospital. Part V. Virological and serological observations. S Afr Med J 68 :733–736.
-
10
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.
-
11
Honig JE, Osborne JC, Nichol ST, 2004. Crimean-Congo hemorrhagic fever virus genome L RNA segment and encoded protein. Virology 321 :29–35.
-
12
Kinsella E, Martin SG, Grolla A, Czub M, Feldmann H, Flick R, 2004. Sequence determination of the Crimean-Congo hemorrhagic fever virus L segment. Virology 321 :23–28.
-
13
Deyde VM, Khristova ML, Rollin PE, Ksiazek TG, Nichol ST, 2006. Crimean-Congo hemorrhagic fever virus genomics and global diversity. J Virol 80 :8834–8842.
-
14
Hewson R, Gmyl A, Gmyl L, Smirnova SE, Karganova G, Jamil B, Hasan R, Chamberlain J, Clegg C, 2004. Evidence of segment reassortment in Crimean-Congo haemorrhagic fever virus. J Gen Virol 85 :3059–3070.
-
15
Whitehouse CA, 2004. Crimean-Congo hemorrhagic fever. Antiviral Res 64 :145–160.
-
16
Yashina L, Petrova I, Seregin S, Vyshemirskii O, Lvov D, Aristova V, Kuhn J, Morzunov S, Gutorov V, Kuzina I, Tyunnikov G, Netesov S, Petrov V, 2003. Genetic variability of Crimean-Congo haemorrhagic fever virus in Russia and Central Asia. J Gen Virol 84 :1199–1206.
-
17
Drosten C, Kummerer BM, Schmitz H, Gunther S, 2003. Molecular diagnostics of viral hemorrhagic fevers. Antiviral Res 57 :61–87.
-
18
Burt FJ, Spencer DC, Leman PA, Patterson B, Swanepoel R, 1996. Investigation of tick-borne viruses as pathogens of humans in South Africa and evidence of Dugbe virus infection in a patient with prolonged thrombocytopenia. Epidemiol Infect 116 :353–361.
-
19
Burt FJ, Leman PA, Smith JF, Swanepoel R, 1998. The use of a reverse transcription-polymerase chain reaction for the detection of viral nucleic acid in the diagnosis of Crimean-Congo haemorrhagic fever. J Virol Methods 70 :129–137.
-
20
Duh D, Saksida A, Petrovec M, Dedushaj I, Avsic-Zupanc T, 2006. Novel one-step real-time RT-PCR assay for rapid and specific diagnosis of Crimean-Congo hemorrhagic fever encountered in the Balkans. J Virol Methods 133 :175–179.
-
21
Yapar M, Aydogan H, Pahsa A, Besirbellioglu BA, Bodur H, Basustaoglu AC, Guney C, Avci IY, Sener K, Setteh MH, Kubar A, 2005. Rapid and quantitative detection of Crimean-Congo hemorrhagic fever virus by one-step real-time reverse transcriptase-PCR. Jpn J Infect Dis 58 :358–362.
-
22
Afonina I, Zivarts M, Kutyavin I, Lukhtanov E, Gamper H, Meyer RB, 1997. Efficient priming of PCR with short oligonucleotides conjugated to a minor groove binder. Nucleic Acids Res 25 :2657–2660.
-
23
Afonina IA, Reed MW, Lusby E, Shishkina IG, Belousov YS, 2002. Minor groove binder-conjugated DNA probes for quantitative DNA detection by hybridization-triggered fluorescence. Biotechniques 32: 940–944, 946–949.
-
24
Saidi S, Casals J, Faghih MA, 1975. Crimean hemorrhagic fever-Congo (CHF-C) virus antibodies in man, and in domestic and small mammals, in Iran. Am J Trop Med Hyg 24 :353–357.
-
25
Kulesh DA, Baker RO, Loveless BM, Norwood D, Zwiers SH, Mucker E, Hartmann C, Herrera R, Miller D, Christensen D, Wasieloski LP Jr, Huggins J, Jahrling PB, 2004. Smallpox and pan-orthopox virus detection by real-time 3′-minor groove binder TaqMan assays on the roche LightCycler and the Cepheid smart Cycler platforms. J Clin Microbiol 42 :601–609.
-
26
Yashina L, Vyshemirskii O, Seregin S, Petrova I, Samokhvalov E, Lvov D, Gutorov V, Kuzina I, Tyunnikov G, Tang YW, Netesov S, Petrov V, 2003. Genetic analysis of Crimean-Congo hemorrhagic fever virus in Russia. J Clin Microbiol 41 :860–862.
-
27
Belousov YS, Welch RA, Sanders S, Mills A, Kulchenko A, Dempcy R, Afonina IA, Walburger DK, Glaser CL, Yadavalli S, Vermeulen NM, Mahoney W, 2004. Single nucleotide polymorphism genotyping by two colour melting curve analysis using the MGB Eclipse Probe System in challenging sequence environment. Hum Genomics 1 :209–217.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 2038 | 1883 | 26 |
| Full Text Views | 315 | 12 | 0 |
| PDF Downloads | 117 | 11 | 0 |
Development of a TaqMan®–Minor Groove Binding Protein Assay for the Detection and Quantification of Crimean-Congo Hemorrhagic Fever Virus
A. R. Garrison
A. R. Garrison
Virology Division and Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; Institute of Virology, Ministry of Health, Tashkent, Uzbekistan
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S. Alakbarova
S. Alakbarova
Virology Division and Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; Institute of Virology, Ministry of Health, Tashkent, Uzbekistan
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D. A. Kulesh
D. A. Kulesh
Virology Division and Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; Institute of Virology, Ministry of Health, Tashkent, Uzbekistan
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D. Shezmukhamedova
D. Shezmukhamedova
Virology Division and Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; Institute of Virology, Ministry of Health, Tashkent, Uzbekistan
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S. Khodjaev
S. Khodjaev
Virology Division and Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; Institute of Virology, Ministry of Health, Tashkent, Uzbekistan
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T. P. Endy
T. P. Endy
Virology Division and Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; Institute of Virology, Ministry of Health, Tashkent, Uzbekistan
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J. Paragas
J. Paragas
Virology Division and Diagnostic Systems Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; Institute of Virology, Ministry of Health, Tashkent, Uzbekistan
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Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus of the genus Nairovirus and the family Bunyaviridae. It is a negative-strand RNA virus comprised of small (S), medium (M), and large (L) genome segments. The S segment encodes for nucleocapsid protein, the M segment codes for envelope glycoproteins (Gn and Gc), and the L segment codes for the RNA-dependent RNA polymerase. Currently, there are a limited number of methods for rapidly diagnosing CCHFV infections. We developed a real-time, reverse transcription–polymerase chain reaction assay for the rapid detection of CCHFV by using the TaqMan®–minor groove binding protein probe technology. The primers and probes were designed to amplify and detect a region in the S segment of CCHFV that is conserved across multiple strains. The limit of detection of the assay was 10 genome copies of RNA. This primer and probe set was specific to 18 strains of CCHFV tested and did not cross-react with either a DNA panel of 78 organisms or a panel of 28 diverse RNA viruses. This will rapidly and specifically detect CCHFV, and it has been used to detect CCHFV infection in samples from humans, animals, and ticks.
Author Notes
- INTRODUCTION
- MATERIALS AND METHODS
- RT-PCR primers, TaqMan®-MGB probe, and target sequences.
- Viral RNA propagation and RNA isolation.
- Real-time RT-PCR assay.
- DNA and RNA panel evaluations.
- Limit of detection.
- Indirect hemagglutination (HI), agarose gel diffusion and precipitation (AGDP), and complement fixation (CF).
- Sequencing the CCHF S segment.
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Swanepoel R, Leman PA, Burt FJ, Jardine J, Verwoerd DJ, Capua I, Bruckner GK, Burger WP, 1998. Experimental infection of ostriches with Crimean-Congo haemorrhagic fever virus. Epidemiol Infect 121 :427–432.
-
2
Dohm DJ, Logan TM, Linthicum KJ, Rossi CA, Turell MJ, 1996. Transmission of Crimean-Congo hemorrhagic fever virus by Hyalomma impeltatum (Acari:Ixodidae) after experimental infection. J Med Entomol 33 :848–851.
-
3
Rodriguez LL, Maupin GO, Ksiazek TG, Rollin PE, Khan AS, Schwarz TF, Lofts RS, Smith JF, Noor AM, Peters CJ, Nichol ST, 1997. Molecular investigation of a multisource outbreak of Crimean-Congo hemorrhagic fever in the United Arab Emirates. Am J Trop Med Hyg 57 :512–518.
-
4
Swanepoel R, Shepherd AJ, Leman PA, Shepherd SP, McGillivray GM, Erasmus MJ, Searle LA, Gill DE, 1987. Epidemiologic and clinical features of Crimean-Congo hemorrhagic fever in southern Africa. Am J Trop Med Hyg 36 :120–132.
-
5
Burt FJ, Leman PA, Abbott JC, Swanepoel R, 1994. Serodiagnosis of Crimean-Congo haemorrhagic fever. Epidemiol Infect 113 :551–562.
-
6
Hoogstraal H, 1979. The epidemiology of tick-borne Crimean-Congo hemorrhagic fever in Asia, Europe, and Africa. J Med Entomol 15 :307–417.
-
7
Burney MI, Ghafoor A, Saleen M, Webb PA, Casals J, 1980. Nosocomial outbreak of viral hemorrhagic fever caused by Crimean Hemorrhagic fever-Congo virus in Pakistan, January 1976. Am J Trop Med Hyg 29 :941–947.
-
8
Papa A, Bino S, Llagami A, Brahimaj B, Papadimitriou E, Pavlidou V, Velo E, Cahani G, Hajdini M, Pilaca A, Harxhi A, Antoniadis A, 2002. Crimean-Congo hemorrhagic fever in Albania, 2001. Eur J Clin Microbiol Infect Dis 21 :603–606.
-
9
Shepherd AJ, Swanepoel R, Shepherd SP, Leman PA, Blackburn NK, Hallett AF, 1985. A nosocomial outbreak of Crimean-Congo haemorrhagic fever at Tygerberg Hospital. Part V. Virological and serological observations. S Afr Med J 68 :733–736.
-
10
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.
-
11
Honig JE, Osborne JC, Nichol ST, 2004. Crimean-Congo hemorrhagic fever virus genome L RNA segment and encoded protein. Virology 321 :29–35.
-
12
Kinsella E, Martin SG, Grolla A, Czub M, Feldmann H, Flick R, 2004. Sequence determination of the Crimean-Congo hemorrhagic fever virus L segment. Virology 321 :23–28.
-
13
Deyde VM, Khristova ML, Rollin PE, Ksiazek TG, Nichol ST, 2006. Crimean-Congo hemorrhagic fever virus genomics and global diversity. J Virol 80 :8834–8842.
-
14
Hewson R, Gmyl A, Gmyl L, Smirnova SE, Karganova G, Jamil B, Hasan R, Chamberlain J, Clegg C, 2004. Evidence of segment reassortment in Crimean-Congo haemorrhagic fever virus. J Gen Virol 85 :3059–3070.
-
15
Whitehouse CA, 2004. Crimean-Congo hemorrhagic fever. Antiviral Res 64 :145–160.
-
16
Yashina L, Petrova I, Seregin S, Vyshemirskii O, Lvov D, Aristova V, Kuhn J, Morzunov S, Gutorov V, Kuzina I, Tyunnikov G, Netesov S, Petrov V, 2003. Genetic variability of Crimean-Congo haemorrhagic fever virus in Russia and Central Asia. J Gen Virol 84 :1199–1206.
-
17
Drosten C, Kummerer BM, Schmitz H, Gunther S, 2003. Molecular diagnostics of viral hemorrhagic fevers. Antiviral Res 57 :61–87.
-
18
Burt FJ, Spencer DC, Leman PA, Patterson B, Swanepoel R, 1996. Investigation of tick-borne viruses as pathogens of humans in South Africa and evidence of Dugbe virus infection in a patient with prolonged thrombocytopenia. Epidemiol Infect 116 :353–361.
-
19
Burt FJ, Leman PA, Smith JF, Swanepoel R, 1998. The use of a reverse transcription-polymerase chain reaction for the detection of viral nucleic acid in the diagnosis of Crimean-Congo haemorrhagic fever. J Virol Methods 70 :129–137.
-
20
Duh D, Saksida A, Petrovec M, Dedushaj I, Avsic-Zupanc T, 2006. Novel one-step real-time RT-PCR assay for rapid and specific diagnosis of Crimean-Congo hemorrhagic fever encountered in the Balkans. J Virol Methods 133 :175–179.
-
21
Yapar M, Aydogan H, Pahsa A, Besirbellioglu BA, Bodur H, Basustaoglu AC, Guney C, Avci IY, Sener K, Setteh MH, Kubar A, 2005. Rapid and quantitative detection of Crimean-Congo hemorrhagic fever virus by one-step real-time reverse transcriptase-PCR. Jpn J Infect Dis 58 :358–362.
-
22
Afonina I, Zivarts M, Kutyavin I, Lukhtanov E, Gamper H, Meyer RB, 1997. Efficient priming of PCR with short oligonucleotides conjugated to a minor groove binder. Nucleic Acids Res 25 :2657–2660.
-
23
Afonina IA, Reed MW, Lusby E, Shishkina IG, Belousov YS, 2002. Minor groove binder-conjugated DNA probes for quantitative DNA detection by hybridization-triggered fluorescence. Biotechniques 32: 940–944, 946–949.
-
24
Saidi S, Casals J, Faghih MA, 1975. Crimean hemorrhagic fever-Congo (CHF-C) virus antibodies in man, and in domestic and small mammals, in Iran. Am J Trop Med Hyg 24 :353–357.
-
25
Kulesh DA, Baker RO, Loveless BM, Norwood D, Zwiers SH, Mucker E, Hartmann C, Herrera R, Miller D, Christensen D, Wasieloski LP Jr, Huggins J, Jahrling PB, 2004. Smallpox and pan-orthopox virus detection by real-time 3′-minor groove binder TaqMan assays on the roche LightCycler and the Cepheid smart Cycler platforms. J Clin Microbiol 42 :601–609.
-
26
Yashina L, Vyshemirskii O, Seregin S, Petrova I, Samokhvalov E, Lvov D, Gutorov V, Kuzina I, Tyunnikov G, Tang YW, Netesov S, Petrov V, 2003. Genetic analysis of Crimean-Congo hemorrhagic fever virus in Russia. J Clin Microbiol 41 :860–862.
-
27
Belousov YS, Welch RA, Sanders S, Mills A, Kulchenko A, Dempcy R, Afonina IA, Walburger DK, Glaser CL, Yadavalli S, Vermeulen NM, Mahoney W, 2004. Single nucleotide polymorphism genotyping by two colour melting curve analysis using the MGB Eclipse Probe System in challenging sequence environment. Hum Genomics 1 :209–217.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 2038 | 1883 | 26 |
| Full Text Views | 315 | 12 | 0 |
| PDF Downloads | 117 | 11 | 0 |