Abstract
Mycoplasma pneumoniae is a significant cause of respiratory disease, accounting for approximately 20% of cases of community-acquired pneumonia. Although several diagnostic methods exist to detect M. pneumoniae in respiratory specimens, real-time PCR has emerged as a significant improvement for the rapid diagnosis of this pathogen. The method described herein details the procedure for the detection of M. pneumoniae by real-time PCR (qPCR). The qPCR assay described can be performed with three targets specific for M. pneumoniae (Mp181, Mp3, and Mp7) and one marker for the detection of the RNaseP gene found in human nucleic acid as an internal control reaction. Recent studies have demonstrated the ability of this procedure to reliably identify this agent and facilitate the timely recognition of an outbreak.
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References
Walter ND, Grant GB, Bandy U, Alexander NE, Winchell JM, Jordan HT, Sejvar JJ, Hicks LA, Gifford DR, Alexander NT, Thurman KA, Schwartz SB, Dennehy PH, Nino K, Fields BS, Dillon MT, Erdman DD, Whitney CG, Moore MR (2008) Community outbreak of Mycoplasma pneumoniae: school-based cluster of neurologic disease associated with household transmission of respiratory illness. J Infect Dis 198:1365–1374
Klement E, Talkington DF, Wasserzug O, Kayouf R, Davidovitch N, Dumke R, Bar-Zeev Y, Merav R, Boxman J, Thacker WL, Wolf D, Lazarovich T, Shemer-Avni Y, Glikman D, Jacobs E, Grotto I, Block C, Nir-Paz R (2006) Identification of risk factors for infection in an outbreak of Mycoplasma pneumoniae respiratory tract disease. Clin Infect Dis 43:1239–1245
Mezarina KB, Huffmire A, Downing J, Core N, Gershman K, Hoffman R (2001) Outbreak of community-acquired pneumonia caused by Mycoplasma pneumoniae—Colorado 2000. MMWR 50:227–230
Waring AL, Halse TA, Csiza CK, Carlyn CJ, Musser KA, Limberger RJ (2001) Development of a genomics-based PCR assay for detection of Mycoplasma pneumoniae in a large outbreak in New York State. J Clin Microbiol 39:1385–1390
Waites KB, Talkington DF (2004) Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev 17:697–728
Waites KB, Balish MF, Atkinson TP (2008) New insights into the pathogenesis and detection of Mycoplasma pneumoniae infections. Future Microbiol 3:635–648
Hammerschlag MR (2001) Mycoplasma pneumoniae infections. Curr Opin Infect Dis 14:181–186
Winchell JM, Thurman KA, Mitchell SL, Thacker WL, Fields BS (2008) Evaluation of three real-time PCR assays for the detection of Mycoplasma pneumoniae in an outbreak investigation. J Clin Microbiol 46:3116–3118
Ursi D, Dirven K, Loens K, Ieven M, Goossens H (2003) Detection of Mycoplasma pneumoniae in respiratory samples by real-time PCR using an inhibition control. J Microbiol Methods 55:149–153
Loens K, Leven M, Ursi D, Beck T, Overdijk M, Sillekens P, Goossens H (2003) Detection of Mycoplasma pneumoniae by real-time nucleic acid sequence-based amplification. J Clin Microbiol 41:4448–4450
Thurman KA, Walter ND, Schwartz SB, Mitchell SL, Dillon MT, Deutscher M, Fulton JP, Tongren JE, Hicks LA, Winchell JM (2008) Comparison of laboratory diagnostic procedures for detection of Mycoplasma pneumoniae in community outbreaks. Clin Infect Dis 48:1244–1249
Kannan TR, Baseman JB (2006) ADP-ribosylating and vacuolating cytotoxin of Mycoplasma pneumoniae represents unique virulence determinant among bacterial pathogens. Proc Natl Acad Sci USA 103:6724–6729
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Winchell, J.M., Mitchell, S.L. (2013). Detection of Mycoplasma pneumoniae by Real-Time PCR. In: Wilks, M. (eds) PCR Detection of Microbial Pathogens. Methods in Molecular Biology, vol 943. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-353-4_10
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DOI: https://doi.org/10.1007/978-1-60327-353-4_10
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