In situ amplification of single copy gene segments in individual cells by the polymerase chain reaction

Infection ◽  
1991 ◽  
Vol 19 (4) ◽  
pp. 242-244 ◽  
Author(s):  
W. Spann ◽  
Katharina Pachmann ◽  
Halina Zabnienska ◽  
Andrea Pielmeier ◽  
B. Emmerich
2004 ◽  
Vol 31 (12) ◽  
pp. 1054-1060 ◽  
Author(s):  
Juan Manuel Morillo ◽  
Laura Lau ◽  
Mariano Sanz ◽  
David Herrera ◽  
Conchita Martin ◽  
...  

Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  

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.


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