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. 1997 May;35(5):1203–1208. doi: 10.1128/jcm.35.5.1203-1208.1997

Detection of Colorado tick fever virus by using reverse transcriptase PCR and application of the technique in laboratory diagnosis.

A J Johnson 1, N Karabatsos 1, R S Lanciotti 1
PMCID: PMC232730  PMID: 9114408

Abstract

Colorado tick fever (CTF) virus elicits an acute illness in humans, producing nonspecific flu-like symptoms and a biphasic fever in approximately 50% of patients. The disease is transmitted by the adult Rocky Mountain wood tick (Dermacentor andersoni), and therefore incidence is limited by the habitat and life cycle of that vector. The early symptoms of infection are difficult to distinguish from those of several other agents, especially Rickettsia rickettsii. Serologic testing is usually unable to provide evidence of CTF viral infection during the acute phase because of the late appearance of the various antibodies. Here we report the development and clinical application of a test to diagnose this disease during the acute stages. Oligonucleotide primers to the S2 segment of CTF (Florio) virus were made, and these were used in the amplification of a 528-bp fragment of DNA, transcribed from the double-stranded CTF virus RNA template by reverse transcriptase PCR. RNAs processed from 16 CTF virus isolates yielded similar results when analyzed on agarose gels. These were distinguishable from their antigenic relatives Eyach, S6-14-03, and T5-2092 and from other coltiviruses and an orbivirus but not from the antigenically distinct CTF virus-related isolate 720896. A mouse model demonstrated the utility of this method with whole-blood specimens, and CTF virus was successfully detected in human sera from the initial day of the onset of symptoms to 8 days later. The reverse transcriptase PCR method is a promising tool for the early diagnosis of CTF viral infection, or for ruling out CTF virus as the etiologic agent, in order to facilitate appropriate medical support.

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Selected References

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  1. Akita G. Y., Chinsangaram J., Osburn B. I., Ianconescu M., Kaufman R. Detection of bluetongue virus serogroup by polymerase chain reaction. J Vet Diagn Invest. 1992 Oct;4(4):400–405. doi: 10.1177/104063879200400405. [DOI] [PubMed] [Google Scholar]
  2. Akita G. Y., Glenn J., Castro A. E., Osburn B. I. Detection of bluetongue virus in clinical samples by polymerase chain reaction. J Vet Diagn Invest. 1993 Apr;5(2):154–158. doi: 10.1177/104063879300500202. [DOI] [PubMed] [Google Scholar]
  3. Bodkin D. K., Knudson D. L. Genetic relatedness of Colorado tick fever virus isolates by RNA-RNA blot hybridization. J Gen Virol. 1987 Apr;68(Pt 4):1199–1204. doi: 10.1099/0022-1317-68-4-1199. [DOI] [PubMed] [Google Scholar]
  4. Bowen G. S., Shriner R. B., Pokorny K. S., Kirk L. J., McLean R. G. Experimental Colorado tick fever virus infection in Colorado mammals. Am J Trop Med Hyg. 1981 Jan;30(1):224–229. doi: 10.4269/ajtmh.1981.30.224. [DOI] [PubMed] [Google Scholar]
  5. Brown S. E., Gorman B. M., Tesh R. B., Knudson D. L. Coltiviruses isolated from mosquitoes collected in Indonesia. Virology. 1993 Sep;196(1):363–367. doi: 10.1006/viro.1993.1490. [DOI] [PubMed] [Google Scholar]
  6. Brown S. E., Miller B. R., McLean R. G., Knudson D. L. Co-circulation of multiple Colorado tick fever virus genotypes. Am J Trop Med Hyg. 1989 Jan;40(1):94–101. doi: 10.4269/ajtmh.1989.40.94. [DOI] [PubMed] [Google Scholar]
  7. Burgdorfer W. Tick-borne diseases in the United States: Rocky Mountain spotted fever and Colorado tick fever. A review. Acta Trop. 1977 Jun;34(2):103–126. [PubMed] [Google Scholar]
  8. Calisher C. H., Poland J. D., Calisher S. B., Warmoth L. A. Diagnosis of Colorado tick fever virus infection by enzyme immunoassays for immunoglobulin M and G antibodies. J Clin Microbiol. 1985 Jul;22(1):84–88. doi: 10.1128/jcm.22.1.84-88.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chang G. J., Trent D. W., Vorndam A. V., Vergne E., Kinney R. M., Mitchell C. J. An integrated target sequence and signal amplification assay, reverse transcriptase-PCR-enzyme-linked immunosorbent assay, to detect and characterize flaviviruses. J Clin Microbiol. 1994 Feb;32(2):477–483. doi: 10.1128/jcm.32.2.477-483.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dangler C. A., de Mattos C. A., de Mattos C. C., Osburn B. I. Identifying bluetongue virus ribonucleic acid sequences by the polymerase chain reaction. J Virol Methods. 1990 Jun;28(3):281–292. doi: 10.1016/0166-0934(90)90121-u. [DOI] [PubMed] [Google Scholar]
  11. Emmons R. W., Lennette E. H. Immunofluorescent staining in the laboratory diagnosis of Colorado tick fever. J Lab Clin Med. 1966 Dec;68(6):923–929. [PubMed] [Google Scholar]
  12. Goodpasture H. C., Poland J. D., Francy D. B., Bowen G. S., Horn K. A. Colorado tick fever: clinical, epidemiologic, and laboratory aspects of 228 cases in Colorado in 1973-1974. Ann Intern Med. 1978 Mar;88(3):303–310. doi: 10.7326/0003-4819-88-3-303. [DOI] [PubMed] [Google Scholar]
  13. Holodniy M., Kim S., Katzenstein D., Konrad M., Groves E., Merigan T. C. Inhibition of human immunodeficiency virus gene amplification by heparin. J Clin Microbiol. 1991 Apr;29(4):676–679. doi: 10.1128/jcm.29.4.676-679.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Karabatsos N., Poland J. D., Emmons R. W., Mathews J. H., Calisher C. H., Wolff K. L. Antigenic variants of Colorado tick fever virus. J Gen Virol. 1987 May;68(Pt 5):1463–1469. doi: 10.1099/0022-1317-68-5-1463. [DOI] [PubMed] [Google Scholar]
  15. Kox L. F., Rhienthong D., Miranda A. M., Udomsantisuk N., Ellis K., van Leeuwen J., van Heusden S., Kuijper S., Kolk A. H. A more reliable PCR for detection of Mycobacterium tuberculosis in clinical samples. J Clin Microbiol. 1994 Mar;32(3):672–678. doi: 10.1128/jcm.32.3.672-678.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kuno G., Mitchell C. J., Chang G. J., Smith G. C. Detecting bunyaviruses of the Bunyamwera and California serogroups by a PCR technique. J Clin Microbiol. 1996 May;34(5):1184–1188. doi: 10.1128/jcm.34.5.1184-1188.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lanciotti R. S., Calisher C. H., Gubler D. J., Chang G. J., Vorndam A. V. Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction. J Clin Microbiol. 1992 Mar;30(3):545–551. doi: 10.1128/jcm.30.3.545-551.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sakamoto K., Punyahotra R., Mizukoshi N., Ueda S., Imagawa H., Sugiura T., Kamada M., Fukusho A. Rapid detection of African horsesickness virus by the reverse transcriptase polymerase chain reaction (RT-PCR) using the amplimer for segment 3 (VP3 gene). Arch Virol. 1994;136(1-2):87–97. doi: 10.1007/BF01538819. [DOI] [PubMed] [Google Scholar]
  19. Sexton D. J., Kanj S. S., Wilson K., Corey G. R., Hegarty B. C., Levy M. G., Breitschwerdt E. B. The use of a polymerase chain reaction as a diagnostic test for Rocky Mountain spotted fever. Am J Trop Med Hyg. 1994 Jan;50(1):59–63. [PubMed] [Google Scholar]

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