Diagnosis of Human Viruses by Polymerase Chain Reaction by Jan Albert, Eva Maria Fenyö (auth.), Professor Yechiel

By Jan Albert, Eva Maria Fenyö (auth.), Professor Yechiel Becker, Professor Gholamreza Darai (eds.)

The foundation for the powerful therapy and medication of a sufferer is the fast prognosis of the disorder and its causative agent, that is in line with the research of the medical indicators coupled with laboratory assessments. even if swift enhance­ ments were made within the laboratory prognosis of virus ailments, the neces­ sary isolation of the causative virus from the medical 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 tricky to spot by way of molecular options, whereas viruses which exist within the scientific fabric in low concentrations are much more daunting to establish. lately, the applying of the polymerase chain response (peR) strategy built by way of ok. D. Mullis and specific within the examine through Saiki et al. (1985) resulted in a revolution in virus analysis. The consistent with method used to be swiftly utilized to the prognosis of viruses in scientific fabric. quantity 1 of Frontiers of Virology presents new details at the advan­ tages of using the in keeping 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 resources for Diagnosis of Human Viruses by Polymerase Chain Reaction Technology

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Only 256 bp bands with several nonspecific background bands were observed. with an endpoint of 15 pg of HUT 102 cell DNA, which was nine times more sensitive than that obtained with single PCR. Products of the nested double PCR are shown in Fig. 2 c. 6 pg of HUT 102 cell DNA, which is calculated to contain 28 C. Matsumoto and K. 2a-d. Polyacrylamide gel electrophoresis of polymerase chain reaction (PCR) products. a HUT 102 cell and CEM cell DNAs were amplified for 30 cycles with primers CD and ~.

In each amplification in a-c. 1g of CEM cell DNA plus O. 7 X 10 3 pg. 2 X 10 3 pg. 6 pg of HUT 102 cell DNA (lanes 1-11. respectively) were used as template DNA. d HUT 102 cell and CEM cell DNA was amplified by the nested double PCR with primers CD and ~ in the first amplification stage and with nested primers Q) and @ in the second amplification stage. 1g CEM cell DNA were used as template DNA in each lane about 2 copies of the pX region. Therefore, the sensitivity of the nested double peR was more than 80 times higher than that of the single peR.

The results of IF for HTLV-I antibody coincided completely with the results of the nested double PCR for detection of provirus DNA in PBMC. Therefore, the IF test correlates well with the presence of provirus in PBMC by PCR, and it is considered as the most appropriate confirmatory test routinely available for screening out donated blood infected with HTLV -I. As for PA, a correlation of the PA titer and the amount of proviral DNA in PBMC was observed. However, in most of the low titer group and even in a small number of the high titer group, the presence of proviral DNA was not detected.

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