Key result
Digoxigenin labelled probes achieved comparable sensitivity to biotin labelled probes for viral nucleic acid detection, and are recommended for tissues with endogenous biotin.
Digoxigenin labelled probes provide comparable sensitivity to biotin labelled probes for in situ hybridization and are advantageous in tissues with endogenous biotin.
Animal data support digoxigenin probes for biotin-rich tissues; human validation required before diagnostic adoption.
A number of in situ hybridization protocols using digoxigenin or biotin labelled probes were assessed for viral nucleic acid detection in formalin fixed, paraffin embedded tissue. Single-step detection protocols for biotin labelled probes produced low sensitivity; however, enzyme based one-step detection protocols for digoxigenin probes produced high sensitivity for both RNA and DNA systems. For both probe types, multistep detection protocols produced equally high sensitivity. Use of an enhanced APAAP procedure for digoxigenin labelled probes achieved maximal sensitivity without use of biotin-strep-tavidin reactions. The sensitivity of nucleic acid detection obtained with a digoxigenin labelled probe is comparable to that obtained using biotin. Digoxigenin labelled probes for nucleic acid detection are recommended for tissues with endogenous biotin.
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McQuaid et al. (1995) studied Viral nucleic acid detection in formalin fixed, paraffin embedded tissue. Digoxigenin labelled DNA and RNA probes vs. Biotin labelled DNA and RNA probes was evaluated on Sensitivity of viral nucleic acid detection. Digoxigenin labelled probes achieved comparable sensitivity to biotin labelled probes for viral nucleic acid detection, and are recommended for tissues with endogenous biotin.
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