High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

Author:

Hacker G. W.,Zehbe I.,Hainfeld J.,Graf A.-H.,Hauser-Kronberger C.,Schiechl A.,Su H.,Dietze O.

Abstract

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

Publisher

Cambridge University Press (CUP)

Subject

General Medicine

Reference13 articles.

1. 13. This work was funded by the Medical Research Society of Salzburg, Austria, and the Lions Cancer Foundation, Uppsala, Sweden.

2. Immunogold-Silver Staining (IGSS) for Detection of Antigenic Sites and DNA Sequences

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