Chemiluminescent in situ hybridization for the detection of cytomegalovirus DNA.

摘要: A chemiluminescent in situ hybridization assay that could combine the sensitivity of substrates, specificity digoxigenin-labeled probes, and spatial morphological resolution localization signal was developed for detection cytomegalovirus (CMV) DNA. CMV DNA cultured CMV-infected cells different clinical samples (tissue sections cellular smears) detected using probes constructed our laboratory were immunoenzymatically visualized employing anti-digoxigenin Fab fragments labeled with alkaline phosphatase adamantil-1,2-dioxetane phenyl phosphate substrate phosphatase. The luminescent from hybrid formation detected, analyzed, measured a high performance, low light level imaging luminograph apparatus connected to an optical microscope personal computer quantitative image analysis. Increasing values emitted photons per second infected cell, corresponding presence hybridized DNA, be found fixed at various times after infection, following replication cycle. When performed on patients acute infections, all positive tested, both frozen paraffin-embedded tissue sections, proving specific sensitive. this work can useful tool sensitive diagnosis viral infection easily adapted detect study any gene sequence inside cells. may also promising estimation quantification nucleic acids present or smears expression