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Journal of Clinical Microbiology, September 2003, p. 4448-4450, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4448-4450.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection of Mycoplasma pneumoniae by Real-Time Nucleic Acid Sequence-Based Amplification

K. Loens,1* M. Ieven,1 D. Ursi,1 T. Beck,1 M. Overdijk,2 P. Sillekens,2 and H. Goossens1

Department of Medical Microbiology, University of Antwerp UIA, Antwerp, Belgium,1 bioMérieux, Boxtel, The Netherlands2

Received 24 February 2003/ Returned for modification 17 April 2003/ Accepted 16 June 2003

Real-time isothermal nucleic acid sequence-based amplification (RT-NASBA) was applied to the detection of Mycoplasma pneumoniae. In vitro-generated M. pneumoniae RNA was used to assess the sensitivity of the assay. The 95% hit rate was 148 molecules of M. pneumoniae RNA in the amplification and 104 molecules of in vitro-generated RNA after nucleic acid extraction. The sensitivity of the RT-NASBA and the conventional NASBA assays corresponded to 5 color-changing units (CCU) of M. pneumoniae. In spiked throat swabs, nasopharyngeal aspirates, bronchoalveolar lavages, and sputum, the sensitivity of both NASBA assays corresponded to 5 to 50 CCU of M. pneumoniae. A total of 17 clinical specimens positive for M. pneumoniae by PCR were also positive by conventional NASBA, but one specimen was negative by RT-NASBA. These results indicate that the sensitivity of detection of M. pneumoniae by RT-NASBA in respiratory samples might be slightly reduced compared to that by conventional NASBA. However, the real-time assay is superior in speed and ease of handling.

* Corresponding author. Mailing address: Department of Medical Microbiology, University of Antwerp, Universiteitsplein 1 S3, B-2610 Wilrijk, Belgium. Phone: 32 3 820 25 51. Fax: 32 3 820 26 63. E-mail: katherine.loens{at}ua.ac.be.

Journal of Clinical Microbiology, September 2003, p. 4448-4450, Vol. 41, No. 9
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.9.4448-4450.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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