Abstract
Polymerase chain reaction is a useful technique in microbial diagnostics to detect and quantify DNA or RNA of low abundance.
Bacterial and viral nucleic acid can be amplified by PCR upon clinical sample extraction using specific primers for classical qualitative PCR and primers and probes for real-time PCR.
Here we describe the Scorpion-probe real-time PCR-based assay that offers thermodynamic advantages due to its kinetic reaction and provides faster performances compared to a classical double-labeled probe-based assays.
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References
Blasi F, Tarsia P, Aliberti S et al (2005) Chlamydia pneumoniae and Mycoplasma pneumoniae. Semin Respir Crit Care Med 26:617–624
Talkington DF, Shott S, Fallon MT et al (2004) Analysis of eight commercial enzyme immunoassay tests for detection of antibodies to Mycoplasma pneumoniae in human serum. Clin Diagn Lab Immunol 11:862–867
Mackay IM (2004) Real-time PCR in the microbiology laboratory. Clin Microbiol Infect 10:190–212
Welti M, Jaton K, Altwegg M et al (2003) Development of a multiplex real-time quantitative PCR assay to detect Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae in respiratory tract secretions. Diagn Microbiol Infect Dis 45:85–95
Stralin K, Backman A, Holmberg H et al (2005) Design of a multiplex PCR for Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae and Chlamydophila pneumoniae to be used on sputum samples. APMIS 113:99–111
Wong ML, Medrano JF (2005) Real-time PCR for mRNA quantitation. Biotechniques 39:75–85
Whitcombe D, Theaker J, Guy SP et al (1999) Detection of PCR products using self-probing amplicons and fluorescence. Nat Biotechnol 17:804–807
Di Marco E, Cangemi G, Filippetti M et al (2007) Development and clinical validation of a real-time PCR using uni-molecular Scorpion-based probe for the detection of Mycoplasma pneumoniae in clinical isolates. New Microbiol 30:415–421
Jie Z (2006) Spectroscopy-based quantitative fluorescence resonance energy transfer analysis. In: Stockand JD, Shapiro MS (eds) Ion channels: methods and protocols, methods in molecular biology, vol 337. Humana, Totowa, NJ, pp 65–77
Zuker M (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31:3406–3415
SantaLucia J Jr (1998) A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics. Proc Natl Acad Sci U S A 95:1460–1465
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Di Marco, E. (2014). Real-Time PCR Detection of Mycoplasma pneumoniae in the Diagnosis of Community-Acquired Pneumonia. In: Biassoni, R., Raso, A. (eds) Quantitative Real-Time PCR. Methods in Molecular Biology, vol 1160. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0733-5_9
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DOI: https://doi.org/10.1007/978-1-4939-0733-5_9
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0732-8
Online ISBN: 978-1-4939-0733-5
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