AIDS demential complex is unexplained in terms of a correlation between the presence of HIV-1 in the brain and the severity of the demential. The objective is to develop simple and reproducible methods for the polymerase chain reaction (PCR) amplification and detection in situ of HIV-1 DNA and RNA (Genomic & mRNA) in post mortem formalin fixed human tissues. With this method, the precise localization of HIV-1 infection and expression patterns can be assigned to specific cell types. Standard oligonucleotide primers for amplification, that discriminate between HIV-1 DNA, and viral messenger RNA's, are to be developed which recognize conserved regions of the virus, thus allowing maximum flexibility in detecting many strains of HIV-1, without sacrificing specificity. The in situ amplification methodology shall be developed with the sensitivity to detect single-copy genes and rare messenger RNA species. Various tissue preservation methods are to be tested for maintenance of morphology after thermocycling amplification in order to allow post-amplification immunohistochemical assignment of cell-type within the brain tissue. Since combining in situ PCR amplification with immunocytochemistry has been cumbersome and difficult for research laboratories to perform with success, it is anticipated that the development of these methods will provide unprecedented technical innovation that will find widespread use throughout the research and diagnostic communities.
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