Open AccessApplicazioni della PCR e PCR in situ nella diagnosi di infezioni batteriche e virali da biopsie fissate in formalina e incluse in paraffina
Author(s) -
S. Cazzavillan,
Emanuela Bonoldi,
Chiara Segala,
Mariella Rossi,
Emanuele D’Amore,
Mario Rassu
Publication year - 2003
Publication title -
microbiologia medica
Language(s) - English
Resource type - Journals
eISSN - 2280-6423
pISSN - 1120-0146
DOI - 10.4081/mm.2003.3060
Subject(s) - biology , in situ , amplicon , primer (cosmetics) , polymerase chain reaction , fastidious organism , microbiology and biotechnology , virology , genetics , chemistry , bacteria , gene , organic chemistry
In situ PCR, amplification of target DNA sequences in fixed cells, is a very useful molecular biology tecnique with potential to combine the high sensitivity of tube PCR with the precise anatomical localization of the targeted bsequences. It allows the study of low copy viral or bacterial DNA. In this study we document the utility of directin situ PCR with single primer pair by applying it to 3 infectious agents in different model systems: Borrelia burgdorferi in 5 Eritema migrans lesions and 55 primitive cutaneous B cell lymphomas, Chlamydia pneumoniae in 200 autoptic atheromasic lesions, and Papilloma virus in 20 CIN 1 (mild cervical dysplasia). In situ PCR seems to be a very promising tecnique; however, the prerequisite for the success of in situ PCR is conditioned by optimal standardization of the key variables which, on the other hand, are influenced by tissue composition