Detection of individual microbial pathogens by proximity ligation
2006 (English)In: Clinical Chemistry, ISSN 0009-9147, E-ISSN 1530-8561, Vol. 52, no 6, 1152-1160 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
2006. Vol. 52, no 6, 1152-1160 p.
Animals, Antibodies; Monoclonal, Bacterial Proteins/analysis/genetics, Bacteriological Techniques, Biotinylation, Enzyme-Linked Immunosorbent Assay, Feces/microbiology, Female, Fetus/virology, Lawsonia Bacteria/*classification/genetics/immunology, Mice, Mice; Inbred BALB C, Nucleic Acid Amplification Techniques, Parvoviridae Infections/veterinary/virology, Parvovirus; Porcine/*classification/genetics/immunology, Pregnancy, Pregnancy Complications; Infectious/veterinary/virology, Sensitivity and Specificity, Swine, Swine Diseases/virology, Viral Proteins/analysis/genetics, Virion/classification/genetics, Virology/methods
IdentifiersURN: urn:nbn:se:uu:diva-94310DOI: 10.1373/clinchem.2005.065847PubMedID: 16723682OAI: oai:DiVA.org:uu-94310DiVA: diva2:168118