Abstract
We developed a loop-mediated isothermal amplification (LAMP) method to detect Chlamydophila pneumoniae infection. This assay exclusively amplified C. pneumoniae sequences and no cross-reactivity was observed for other Chlamydia species. The detection limit for this assay was found to be ten elementary bodies in 25 min, as observed in a real-time turbidimeter and electrophoretic analysis. The specificity of the LAMP reaction was confirmed by restriction endonuclease analysis, as well as direct sequencing of the amplified product. Among nasopharyngeal swab specimens from 120 patients with acute respiratory tract infections and 40 healthy individuals, the LAMP results showed 100% agreement with the results of real-time polymerase chain reaction (PCR) assays.
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Acknowledgments
The authors wish to thank Masako Endou and Asami Okazaki, Department of Pediatrics, Kawasaki Medical School, Japan, for their technical assistance. This work was supported by the MEXT KAKENHI (19591190) and Project Research Grants from Kawasaki Medical School (13–401, 14–402, 15–405A, 16–405M, 17–402M, 18–401, 19–402M, and 20–4030).
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Kawai, Y., Miyashita, N., Kishi, F. et al. Development and evaluation of a loop-mediated isothermal amplification method for the rapid detection of Chlamydophila pneumoniae . Eur J Clin Microbiol Infect Dis 28, 801–805 (2009). https://doi.org/10.1007/s10096-009-0710-z
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DOI: https://doi.org/10.1007/s10096-009-0710-z