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.
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.
Authors: Nithya Jambunathan; Anu-Susan Charles; Ramesh Subramanian; Ahmad A Saied; Misagh Naderi; Paul Rider; Michal Brylinski; Vladimir N Chouljenko; Konstantin G Kousoulas Journal: J Virol Date: 2015-12-09 Impact factor: 5.103
Authors: James Flanigon; Masood Kamali-Moghaddam; Ian Burbulis; Carla Annink; Martin Steffen; Paul Oeth; Roger Brent; Dirk van den Boom; Ulf Landegren; Charles Cantor Journal: N Biotechnol Date: 2012-11-28 Impact factor: 5.079
Authors: Sigrun M Gustafsdottir; Stefan Wennström; Simon Fredriksson; Edith Schallmeiner; Andrew D Hamilton; Said M Sebti; Ulf Landegren Journal: Clin Chem Date: 2008-05-22 Impact factor: 8.327
Authors: Michael S Akhras; Sreedevi Thiyagarajan; Andrea C Villablanca; Ronald W Davis; Pål Nyrén; Nader Pourmand Journal: PLoS One Date: 2007-02-21 Impact factor: 3.240