Detection of Viral Sequences in Formalin Fixed, Paraffin Embedded Tissues from HIV-1 Infected Patients Using the PCR
The development of new techniques can further the elucidation of the biology of many diseases. The invention of the polymerase chain reaction (PCR) in the 1980’s by Mullis and co-workers at the Cetus Corporation (Saiki et al. 1985) has revolutionized the field of molecular virology. The ability of the PCR to detect as few as one target present among 100,000 to 1,000,000 human cells has permitted the direct examination of in vivo viral infections. Prior to the PCR, extremely low numbers of virus were often “invisible” by many techniques. Culture techniques were generally the most sensitive method of direct viral detection. However, it was theoretically possible that some latent viruses would not grow in culture. Immunohistochemistry and in situ hybridization techniques can detect the expression of viral antigens or RNA, or the presence of multiple viral genomes. Unfortunately, these techniques suffer from various drawbacks. In particular, latent viral infection with small numbers of viral genomes (generally less than 50 copies per cell) would escape detection. The Southern blot technique can detect viral DNA regardless of its state of activation, but generally needs at least one viral genome present per 100 cells for detection.
KeywordsFormalin Lymphoma Leukemia Electrophoresis Anemia
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