|
|
|
|
 |
|||
|
|||
Detection protocols for biotinylated probes: optimization using multistep techniques
S McQuaid and GM Allan
Neuropathology Laboratory, Queen's University Belfast, Royal Victoria Hospital, Northern Ireland.
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.
Volume 40,
Issue 4,
pp. 569-574,
04/01/1992
Copyright © 1992 by The Histochemical Society
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Divjak, E. M. Glare, and E. H. Walters Improvement of Non-radioactive In Situ Hybridization in Human Airway Tissues: Use of PCR-generated Templates for Synthesis of Probes and an Antibody Sandwich Technique for Detection of Hybridization J. Histochem. Cytochem., April 1, 2002; 50(4): 541 - 548. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. W. Said, M. R. Rettig, K. Heppner, R. A. Vescio, G. Schiller, H. J. Ma, D. Belson, A. Savage, I. P. Shintaku, H. P. Koeffler, et al. Localization of Kaposi's Sarcoma-Associated Herpesvirus in Bone Marrow Biopsy Samples From Patients With Multiple Myeloma Blood, December 1, 1997; 90(11): 4278 - 4282. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. F. Schmidt, J. Chao, Z. Zhu, R. L. DeBiasio, and G. Fisher Signal Amplification in the Detection of Single-copy DNA and RNA by Enzyme-catalyzed Deposition (CARD) of the Novel Fluorescent Reporter Substrate Cy3.29-Tyramide J. Histochem. Cytochem., March 1, 1997; 45(3): 365 - 374. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. E. Martone, J. A. Pollock, Y. Z. Jones, and M. H. Ellisman Ultrastructural Localization of Dendritic Messenger RNA in Adult Rat Hippocampus J. Neurosci., December 1, 1996; 16(23): 7437 - 7446. [Abstract] [Full Text] [PDF]
|
|
| Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact |