By Jan Albert, Eva Maria Fenyö (auth.), Professor Yechiel Becker, Professor Gholamreza Darai (eds.)
The foundation for the powerful therapy and remedy of a sufferer is the fast prognosis of the affliction and its causative agent, that is in line with the research of the scientific signs coupled with laboratory exams. even though quick enhance ments were made within the laboratory prognosis of virus ailments, the neces sary isolation of the causative virus from the scientific specimens is a comparatively lengthy process. Viruses which combine into the mobile DNA (such as human immunodeficiency virus, HIV -1, or hepatitis B virus) are tough to spot via molecular innovations, whereas viruses which exist within the medical fabric in low concentrations are much more daunting to determine. lately, the applying of the polymerase chain response (peR) strategy built by means of ok. D. Mullis and specified within the learn via Saiki et al. (1985) resulted in a revolution in virus prognosis. The consistent with strategy was once quickly utilized to the prognosis of viruses in medical fabric. quantity 1 of Frontiers of Virology presents new details at the advan tages of using the in step with for the prognosis of many human disease-causing viruses, in addition to on a few issues of its use.
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Extra info for Diagnosis of Human Viruses by Polymerase Chain Reaction Technology
Typically, very small amounts of virus are produced; a small fraction of cells contain proviral DNA (estimates of < 1 in 500 to > 1 in 750 have been published) (Rosenblatt et al. 1990; Matsumoto et al. 1990). Serologic Diagnosis and Isolation Numerous methods for serologic diagnosis have been described, and many early reports ofHTLV-I seroprevalence undoubtedly erroneously overestimated the rate of infection with this virus. This occurred because a reliable "confirmatory" test for infection was not available until the late 1980s.
The following is a recommended protocol. Method Synthesis of Oligonucleotides. Oligonucleotides were synthesized on an Applied Biosystems 381A DNA synthesizer by the phosphoramidate method and purified with oligonucleotide purification cartridges (Applied Biosystems). c. 26 Matsumoto and K. Nishioka Source and Isolation of DNA. DNA from two human T-cell lines or PBMC was isolated by the phenol-chloroform method. One of the cell lines, CEM, was uninfected with HTLV-I, and the other, HUT 102, was HTLV-I positive (Poiesz et al.
Methods to distinguish HTLV-I and -II using short synthetic peptide epitopes specific for each virus are under development. In the past, HTLV culture had been difficult because positive cultures produced very little reverse transcriptase; there were thus few convenient and reliable means to monitor in vitro replication of virus. The recent commercial availability of antigen-capture EIA kits has made culture a much more practical option. 40 B. Hjelle Materials and Methods I favor dot-blot hybridization for the detection ofPCR products because of the simplicity and high sensitivity of this method.