Ready to use dry-reagent PCR assays for the four common bacterial pathogens using switchable lanthanide luminescence probe system.
Ready to use dry-reagent PCR assays for Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas spp. and for broad-range bacteria detection were developed. The assays were based on novel switchable lanthanide probes that provide sensitive target DNA detection with exceptionally high signal-to-background ratio, thus enabling clear discrimination between positive and negative results. For example, sensitivity of three S. aureus and two S. pneumonia bacteria (colony forming units) per PCR assay was measured with fluorescence signal more than 30 times over the background signal level. The rapid and easy-to-use assays are suitable for routine clinical diagnostics without molecular biology expertise and facilities.
Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, Finland. Electronic address: artule@utu.fi.
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Lehmusvuori
Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, Finland.
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Soikkeli
Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, Finland.
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Tuunainen
Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, Finland.
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Seppä
Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, Finland.
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Spangar
Clinical Microbiology Laboratory, Turku University Hospital, 20521 Turku, Finland.
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Rantakokko-Jalava
Abacus Diagnostica Oy, 20520 Turku, Finland.
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von Lode
Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, Finland.
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Karhunen
Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, Finland.
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Soukka
Department of Biochemistry/Biotechnology, University of Turku, 20520 Turku, Finland.
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Wittfooth
D001424:Bacterial Infections / Q000175:diagnosis*; D001431:Bacteriological Techniques / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D028581:Lanthanoid Series Elements / Q000378:metabolism*; D008163:Luminescent Measurements; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011549:Pseudomonas / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification
D028581:Lanthanoid Series Elements; D015345:Oligonucleotide Probes
Bacteria detection; Dry-reagent PCR; Switchable lanthanide luminescence