[Application of in situ PCR to clinical pathology--detection of viral infection].
In situ polymerase chain reaction(PCR) combines the sensitivity of solution PCR with the subcellular localization provided by traditional in situ hybridization(ISH). Detection threshold levels for ISH are about 10-20 copies per cell. Therefore rare RNA and DNA sequences are often below detection. PCR is an extremely sensitive technique which amplifies single copy gene sequence to high levels, easily detectable by ISH. Thus, this new technique could have important applications in infectious disease and oncology. Studies on in situ PCR detection of viral DNA sequences include HIV, HPV, CMV, and HBV. The clinical aspects of viral disease have also been studied with a limited number of cases. For example, with HIV, it has been discovered that a larger number of cells previously thought to be uninfected, are latently infected with the virus. In spite of its potentially powerful application for diagnostics, the technique has its own limitation outside research laboratories at present. One of them is a lack of an standardized protocol which can be used for wide range of biological samples. The challenges of this technique and its potential for applications in pathology is discussed.
Aoyagi
K
K
PE Applied Biosystems Division, CA 94404, USA.
D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D017403:In Situ Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology
D004279:DNA, Viral