Molecular Imaging and Biology

, Volume 18, Issue 4, pp 527–534

Detection of Klebsiella. Pneumoniae Infection with an Antisense Oligomer Against its Ribosomal RNA

  • Ling Chen
  • Dengfeng Cheng
  • Guozheng Liu
  • Shuping Dou
  • Yuzhen Wang
  • Xinrong Liu
  • Yuxia Liu
  • Mary Rusckowski
Research Article

Abstract

Purpose

Previously, we demonstrated specific accumulation into bacteria of a 12-mer phosphorodiamidate morpholino (MORF) oligomer complementary to a ribosomal RNA (rRNA) segment found in all bacteria using the universal probe called Eub338 (Eub). Here, two MORF oligomers Eco and Kpn with sequences specific to the rRNA of Escherichia coli (Eco) and Klebsiella pneumoniae (Kpn) were investigated along with Eub and control (nonEub).

Procedures

To determine bacterial rRNA binding, oligomers were tagged with Alexa Fluor 633 (AF633) for fluorescence in situ hybridization (FISH) and fluorescence microscopy, and radiolabeled with technetium-99m (Tc-99m) for biodistribution and SPECT imaging in infected mice.

Results

By both FISH and fluorescence microscopy, Eub showed a positive signal in both E. coli and K. pneumoniae as expected, and Kpn showed significantly higher accumulation in K. pneumoniae with near background in E. coli (p < 0.01). Conversely, Eco was positive in both E. coli and K. pneumoniae, hence nonspecific. As determined by biodistribution, the accumulation of [99mTc]Kpn was higher in the thigh infected with live K. pneumoniae than with live E. coli (p = 0.05), and significantly higher than with heat-killed K. pneumoniae (p = 0.02) in the target thigh. By SPECT imaging, the accumulation of [99mTc]Kpn was obviously higher in its specific target of K. pneumoniae compared to an E. coli infected thigh.

Conclusions

Kpn complementary to the rRNA of K. pneumoniae, labeled with Tc-99m or AF633, demonstrated specific binding to fixed and live K. pneumoniae in culture and in infected mice such that Tc-99m-labeled Kpn as the MORF oligomer may be useful for K. pneumoniae infection detection through imaging.

Key words

Antisense MORF Bacterial ribosomal RNA Bacterial infection imaging 

Copyright information

© World Molecular Imaging Society 2016

Authors and Affiliations

  • Ling Chen
    • 1
  • Dengfeng Cheng
    • 2
  • Guozheng Liu
    • 1
  • Shuping Dou
    • 1
  • Yuzhen Wang
    • 1
  • Xinrong Liu
    • 3
  • Yuxia Liu
    • 4
  • Mary Rusckowski
    • 1
  1. 1.Division of Nuclear Medicine, Department of RadiologyUniversity of Massachusetts Medical SchoolWorcesterUSA
  2. 2.Department of Nuclear Medicine, Zhongshan HospitalFudan UniversityShanghaiChina
  3. 3.ImmunoGen IncWalthamUSA
  4. 4.Shanghai Institute of Applied Physics, Chinese Academy of Sciences, ShanghaiChina

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