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J Nucl Med. 2008; 49 (Supplement 1):161P
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Novel Approaches to Molecular Imaging

Novel Approaches to Molecular Imaging Posters

Successful binding to bacterial ribosomal RNA by a radiolabeled MORF as a potential infection specific imaging agent

Ling Chen1, Donald Hnatowich1 and Mary Rusckowski1

1 Radiology, University of Massachusetts Medical School, Worcester, Massachusetts

703

Objectives: Bacteria share a common ribosomal RNA sequence that could serve as a target for a radiolabeled oligomer with an antisense sequence designed for infection imaging. We are evaluating whether a MORF oliogomer with a sequence specific to a segment of bacterial 16S rRNA can be used to identify bacterial infections through imaging.

Methods: A MORF sequence universal to all bacterial 16S rRNA (Eub338: 5'GCTGCCTCCCGT3') and a control sequence (nonEub338: 5'AGGGCATCCTCA3') were obtained with an amine on the 3’ end for conjugation. The MORFs were conjugated with both the NHS derivatives of Alexa 633 as a fluorescence label and MAG3 for radiolabeling with 99mTc. The study and control MORFs were evaluated in binding studies with Klebsiella (gram negative) and Staphylococcus (gram positive) bacteria. After incubation, bacterial binding of the Alexa 633-MORFs was evaluated by flow cytometry while binding of the 99mTc-MORFs to the RNA fraction was evaluated by isolating the RNA with TRIzol® and counting the total RNA.

Results: By flow cytometry, the study MORF sequence showed an 8 and 80-fold higher binding to Klebsiella and Staphylococcus respectively compared to the control. For reasons not yet established, accumulations of both fluorescent MORFs were 2-fold higher in bacteria Klebsiella than Staphylococcus. In agreement with the flow cytometry results, higher counts were associated with bacteria for the study compared to the control MORF. Isolating and counting total RNA confirmed that this binding was to the target RNA.

Conclusions: A labeled MORF antisense to a segment of bacterial 16S rRNA showed higher binding to bacterial RNA than a control MORF and may be considered for infection specific imaging.





This Article
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Right arrow Articles by Rusckowski, M.
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Right arrow Articles by Chen, L.
Right arrow Articles by Rusckowski, M.