Previous studies showed that Microsatellite DNA Instability is a common genetic alteration in sputum cells of COPD and Asthmatic patients. The aim of this study was to investigate the presence of the phenomenon in nasal aspirates from COPD and Asthmatic patients.
Nasal aspirates and peripheral blood from 3 COPD and 4 asthmatic patients were analyzed. For nasal aspirates 1ml of phosphate buffered saline was inserted directly into nasal cavity via a tapered end syringe and the secretions were aspirated back into the syringe. DNA was extracted using QIAamp tissue and blood kits according to manufacturer’s instructions with some modification. Microsatellite DNA amplification was peformed using primers D14S588, G29802, D6S344, D5S207, D13S71, harboring susceptibility genes for Asthma, Allergy and irreversible airway obstruction. PCR was carried out to amplify Microsatellite DNA sequences. The presence of MSI and/or LOH were scored using a Licor 2400 DNA Sequencer.
Microsatellite Instability and/or Loss of Heterozygosity (LOH) are detectable phenomena in nasal aspirates in humans. LOH was detected at the Asthma group in one Microsatellite marker, namely D14S588 located at chromosome 14 related up to now with a1-antitrypsine gene.
Our preliminary results showed that genetic alterations could easily be detected in nasal aspirates from COPD and Asthmatic patients, at the Microsatellite DNA level.
Moreover, nasal aspirates can be safely and noninvasively obtained. Thus could potentialy be used to evaluate somatic mutations in the upper airways of those patients.
A.D. Karatzanis, None.