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Clinical Chemistry 52: 1152-1160, 2006; 10.1373/clinchem.2005.065847
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(Clinical Chemistry. 2006;52:1152-1160.)
© 2006 American Association for Clinical Chemistry, Inc.

Automation and Analytical Techniques

Detection of Individual Microbial Pathogens by Proximity Ligation

Sigrun M. Gustafsdottir1,2, Ann Nordengrahn2,2, Simon Fredriksson3, Per Wallgren4, Esteban Rivera5, Edith Schallmeiner1, Malik Merza2 and Ulf Landegren1,a

1 The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala, Sweden.
2 Svanova Biotech AB, Uppsala Science Park, Uppsala, Sweden.
3 Stanford Genome Technology Center, Stanford University, Palo Alto, CA.
Departments of4 Pigs, Poultry, and Ruminants and 5 Vaccine Research, National Veterinary Institute, Uppsala, Sweden.

aAddress correspondence to this author at: The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, 75185 Uppsala, Sweden. Fax 46-18-526849; e-mail ulf.landegren{at}genpat.uu.se.

Background: Nucleic acid amplification allows the detection of single infectious agents. Protein-based assays, although they provide information on ongoing infections, have substantially less detection sensitivity.

Methods: We used proximity ligation reactions to detect proteins on bacteria and virus particles via nucleic acid amplification. Antibodies recognizing viral or bacterial surface proteins were equipped with DNA strands that could be joined by ligation when several antibodies were bound in proximity to surface proteins of individual infectious agents.

Results: Detection sensitivities similar to those of nucleic acid-based detection reactions were achieved directly in infected samples for a parvovirus and an intracellular bacterium.

Conclusions: This method enables detection of ligated DNA strands with good sensitivity by real-time PCR and could be of value for early diagnosis of infectious disease and in biodefense.




The following articles in journals at HighWire Press have cited this article:


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S. M. Gustafsdottir, S. Wennstrom, S. Fredriksson, E. Schallmeiner, A. D. Hamilton, S. M. Sebti, and U. Landegren
Use of Proximity Ligation to Screen for Inhibitors of Interactions between Vascular Endothelial Growth Factor A and Its Receptors
Clin. Chem., July 1, 2008; 54(7): 1218 - 1225.
[Abstract] [Full Text] [PDF]


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