A COMPARISON OF DIGOXIGENIN AND BIOTIN-LABELED DNA AND RNA PROBES FORIN-SITU HYBRIDIZATION

A number of in situ hybridization protocols using digoxigenin or bioti n labelled probes were assessed for viral nucleic acid detection in fo rmalin fixed, paraffin embedded tissue. Single-step detection protocol s for biotin labelled probes produced low sensitivity; however, enzyme based one-step detection protocols for digoxigenin probes produced hi gh sensitivity for both RNA and DNA systems. For both probe types, mul tistep detection protocols produced equally high sensitivity. Use of a n enhanced APAAP procedure for digoxigenin rebelled probes acheived ma ximal sensitivity without use of biotin-streptavidin reactions. The se nsitivity of nucleic acid detection obtained with a digoxigenin labell ed probe is comparable to that obtained using biotin. Digoxigenin labe lled probes for nucleic acid detection are recommended for tissues wit h endogenous biotin.