Formalin-Fixed Paraffin-Embedded Tissue (FFPET) Sections for Nucleic Acid-Based Analysis in Biomarker Discovery and Early Drug Development

2015 ◽  
pp. 187-219
Author(s):  
Sabine Lohmann ◽  
Beatrix Bahle ◽  
Andrea Herold ◽  
Julian Schuster
Keyword(s):  
Nucleic Acid ◽  
Drug Development ◽  
Biomarker Discovery ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed ◽  
Early Drug

scholarly journals Detection protocols for biotinylated probes: optimization using multistep techniques.

1992 ◽  
Vol 40 (4) ◽  
pp. 569-574 ◽  
Author(s):  
S McQuaid ◽  
G M Allan
Keyword(s):  
Nucleic Acid ◽  
Measles Virus ◽  
Single Step ◽  
Step Detection ◽  
Formalin Fixed Paraffin ◽  
Wide Range ◽  
Formalin Fixed Paraffin Embedded ◽  
Infected Cells ◽  
Formalin Fixed

Recent studies using biotinylated in situ hybridization (ISH) have utilized a wide range of detection protocols for the biotinylated hybrids, leading to conflicting reports in the literature regarding sensitivity. In this study we compared 11 different detection protocols for biotinylated ISH using a measles virus-specific RNA probe on formalin-fixed, paraffin-embedded central nervous system tissue infected with measles virus. Maximum sensitivity was achieved with five-step detection protocols incorporating the use of a monoclonal antibody to biotin. Single-step detection protocols were found to be insensitive, as shown by their failure to detect viral nucleic acid in infected white-matter cells. Only by increasing the number of steps in the detection protocols were these infected cells demonstrable. Unless pre-hybridization, hybridization, and detection protocols are optimized, the results obtained in pathogenicity studies using ISH could be misinterpreted, leading to false conclusions about nucleic acid distribution. This also applies to the ever-increasing use of ISH for diagnostic purposes.

Sign in / Sign up

Export Citation Format

Share Document