Summary
The detection of human spumaretrovirus (HSRV)-specific RNA and DNA sequences by polymerase chain reactions (PCR) in human cells is described. Various oligodeoxynucleotides of different chain lengths were derived from the prototypic HSRV sequence, synthesized, and employed as sense and antisense primers in several modified versions of a standard PCR. PCR seems to be the method of choice for detecting HSRV nucleic acids and is superior to other techniques. The only serious problem encountered is the oversensitivity of this method that can lead to false-positive results due to laboratory contaminations. Controls and the complete physical separation of all reagents and equipment are required to avoid any contaminations.
This research project was financed by a grant KI 18901/6 from the Bundesmimstermm für Forschung und Technologic
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Achong G, Mansell PWA, Epstein MA, Clifford P (1971) An unusual vims in cultures from a human nasopharyngeal carcinoma. J Natl Cancer Inst 42:299–307
Cullen B (1991) Human immunodeficiency virus as a prototype complex retrovirus. J Virol 65:1053–1056
Davis LG, Dibner MD, Battey JF (1986) Basic methods in molecular biology. Elsevier, New York, pp 79–152
Doolittle RF, Feng D-F, Johnson MS, McClure MA (1989) Origins and evolutionary relationship of retro viruses. Q Rev Biol 64:1–30
Flügel RM (1991) Spumaviruses: a group of complex retroviruses. J AIDS 4:739–750
Flügel RM, Rethwilm A, Maurer B, Darai G (1987) Nucleotide sequence analysis of the env gene and its flanking regions of the human spumaretrovirus reveals two novel genes. EMBO J 6:2077–2084
Frohman MA (1990) Race: rapid amplification of cDNAs ends. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols, a guide to methods and applications. Academic, San Diego, pp 28–53
Kestler HW, Ringler DJ, Mori K, Panicalli DL, Sehgal P, Daniel MD, Desrosiers RC (1991) Importance of the nef gene for maintenance of high virus loads and for development of AIDS. Cell 65:651–662
Kinoshita T, Shimoyana M, Tobinai K, Ito M, Ito S-I, Ikeda S, Tajima K, Shimötono K, Sugimura T (1989) Detection of mRNA for the tax/rex gene of human T-cell leukemia virus type 1 in fresh peripheral blood mononuclear cells of adult T-cell leukemia patients and viral carriers by using polymerase chain reactions. Proc Natl Acad Sci USA 86:5620–5624
Lewe G, Flügel RM (1989) Tracing genetic events in the evolutionary pathway of retroviruses: comparative analysis of the pol and env protein sequences. Virus Genes 2:195–204
Löchelt M, Zentgraf H, Flügel RM (1991) Construction of an infectious DNA clone of the full-length human spumaretrovirus genome and mutagenesis of the bel 1 gene. Virology 184 (in press)
Maurer B, Bannert H, Darai G, Flügel RM (1988) Analysis of the primary structure of the LTR, the gag and the pol gene of the human spumaretrovirus. J Virol 62:1590–1597
Maurer B, Flügel RM (1988) Genomic organization of the human spumaretrovirus and its relatedness to AIDS and other retroviruses. AIDS Res Hum Retroviruses 4:467–473
Muranyi W, Flügel RM (1991) Analysis of splicing patterns by polymerase chain reaction of the human spumaretrovirus reveals complex RNA structures. J Virol 65:727–735
Myers G, Pavlakis GN (1991) Evolutionary potential of complex retroviruses. In: Wagner RR, Frenkel-Conrat H Levy J (ed) The retroviridae: Plenum, New York, ppl-37
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor
Schwartz S, Felber B, Benko DM, Fenyö E-M, Pavlakis G (1990) Cloning and functional analysis of multiply spliced mRNA species of human immunodeficiency virus type 1. J Virol 64:2519–2529
Xiong Y, Eickbush TH (1988) Origin and evolution of retroelements based upon their reverse transcriptase sequences. EMBO J 10:3353–3362
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1992 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Muranyi, W., Flügel, R.M. (1992). Detection of Human Spumaviruses by Polymerase Chain Reaction. In: Becker, Y., Darai, G. (eds) Diagnosis of Human Viruses by Polymerase Chain Reaction Technology. Frontiers of Virology, vol 1. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-84766-0_5
Download citation
DOI: https://doi.org/10.1007/978-3-642-84766-0_5
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-84768-4
Online ISBN: 978-3-642-84766-0
eBook Packages: Springer Book Archive