Detection of Individual Microbial Pathogens by Proximity Ligation

• Published in: Clinical chemistry (Baltimore, Md.) Vol. 52; no. 6; pp. 1152 - 1160
• Main Authors: Gustafsdottir, Sigrun M, Nordengrahn, Ann, Fredriksson, Simon, Wallgren, Per, Rivera, Esteban, Schallmeiner, Edith, Merza, Malik, Landegren, Ulf
• Format: Journal Article
• Language: English
• Published: Washington, DC Am Assoc Clin Chem 01.06.2006; American Association for Clinical Chemistry; Oxford University Press
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Antibodies, Monoclonal; Bacterial Proteins - analysis; Bacterial Proteins - genetics; Bacterial Proteins/analysis/genetics; Bacteriological Techniques; Biological and medical sciences; Biotinylation; Enzyme-Linked Immunosorbent Assay; Feces - microbiology; Female; Fetus - virology; Fundamental and applied biological sciences. Psychology; Investigative techniques, diagnostic techniques (general aspects); Lawsonia Bacteria - classification; Lawsonia Bacteria - genetics; Lawsonia Bacteria - immunology; Lawsonia Bacteria/classification/genetics/immunology; Medical sciences; Mice; Mice, Inbred BALB C; Nucleic Acid Amplification Techniques; Parvoviridae Infections - veterinary; Parvoviridae Infections - virology; Parvoviridae Infections/veterinary/virology; Parvovirus; Parvovirus, Porcine - classification; Parvovirus, Porcine - genetics; Parvovirus, Porcine - immunology; Pregnancy; Pregnancy Complications, Infectious - veterinary; Pregnancy Complications, Infectious - virology; Sensitivity and Specificity; Swine; Swine Diseases - virology; Viral Proteins - analysis; Viral Proteins - genetics; Viral Proteins/analysis/genetics; Virion - classification; Virion - genetics; Virion/classification/genetics; Virology - methods; Human; Proximity; Pathogenic; Clinical biology; Biochemistry; Diagnosis; Molecular biology; Inbred BALB C; Parvovirus; Porcine/classification/genetics/immunology; Infectious/veterinary/virology; Pregnancy Complications; Antibodies; Mice; Monoclonal
• ISSN: 0009-9147; 1530-8561; 1530-8561
• DOI: 10.1373/clinchem.2005.065847

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