A comparison of digoxigenin and biotin labelled DNA and RNA probes for in situ hybridization.
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.
McQuaid
S
S
Neuropathology Laboratory, Queen's University, Belfast, Royal Victoria Hospital, Northern Ireland.
McMahon
J
J
Allan
G M
GM
D000818:Animals; D001710:Biotin; D001921:Brain / Q000821:virology; D002645:Chickens; D004279:DNA, Viral / Q000032:analysis*; D004076:Digoxigenin; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008459:Measles virus / Q000235:genetics / Q000302:isolation & purification; D015336:Molecular Probe Techniques; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity
D004279:DNA, Viral; D012367:RNA, Viral; D001710:Biotin; D004076:Digoxigenin