Effect of the psoralen-based photochemical pathogen inactivation on mitochondrial DNA in platelets.

Abstract:

:Photochemical treatment (PCT) of platelet concentrates, using amotosalen HCl and UVA-light, inactivates pathogens by forming adducts between amotosalen and nucleic acids. The impact of the photochemical treatment on pathogens and leukocytes has been studied extensively. Yet little is known about the effect of PCT on nucleic acids in platelets. Platelets contain viable mitochondria and mitochondrial DNA (mtDNA) and this study aimed at evaluating the amotosalen modifications on platelet mtDNA. We applied two independent but complementary molecular assays to investigate qualitative as well as quantitative aspects of the psoralen-mediated DNA modifications in platelet mtDNA. The amotosalen-DNA modification density was measured using (14)C-labeled amotosalen. Amotosalen (150 microM) yielded 4.0 +/- 1.2 psoralen adducts per 1,000 bp in mtDNA after irradiation with 3 J/cm(2) UVA. Furthermore, we tested if the PCT-induced DNA modifications could be detected by a PCR assay. On the basis of PCR inhibition due to amotosalen-DNA adducts, mtDNA-specific PCR assays were developed and tested for their specificity and sensitivity. Our data revealed that mtDNA in platelets is substantially modified by PCT and that these modifications can be documented by a PCR inhibition system.

journal_name

Platelets

journal_title

Platelets

authors

Bruchmüller I,Lösel R,Bugert P,Corash L,Lin L,Klüter H,Janetzko K

doi

10.1080/09537100500129300

subject

Has Abstract

pub_date

2005-12-01 00:00:00

pages

441-5

issue

8

eissn

0953-7104

issn

1369-1635

pii

M128717716582W4K

journal_volume

16

pub_type

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