[Sensitive and rapid detection of Mycoplasma pneumoniae by loop-mediated isothermal amplification].
For establishment of a method for rapid diagnosis of Mycoplasma pneumoniae in clinical specimens, we designed and evaluated a loop-mediated isothermal amplification (LAMP) assay, using a set of primers targeting the SDC1 repetitive element of the M. pneumoniae genome. The method showed rapid and specific amplification for all the strains of M. pneumoniae tested (Type I and II), within 1 hour at 65 degrees C. No cross-reactivity with the most common causative organisms bacterial pneumonia was observed. The detection limit was shown as 6 copies, which was equal to or higher than that of the two nested PCRs used as references. Two hundred four clinical samples, comprising sputum samples, throat swabs, etc., were tested by both LAMP and PCR, and determination of the correlations revealed complete agreement individually (24 positives). The LAMP assay, a simple procedure in a closed system, allowed rapid amplification and accurate detection consistently; therefore we considered that it could become an caeasily available diagnostic method suitable for clinical situations requiring quick and appropriate decisions for treatments and care.
Eiken Chemical Co, Ltd Biochemical Research Laboratory.
Manabu
M
Yoshino
Toshimitsu
T
Annaka
Tadashi
T
Kojima
Masanari
M
Ikedo
D017714:Community-Acquired Infections / Q000175:diagnosis* / Q000382:microbiology*; D017931:DNA Primers; D016680:Genome, Bacterial / Q000235:genetics; D006801:Humans; D009177:Mycoplasma pneumoniae / Q000235:genetics* / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011019:Pneumonia, Mycoplasma / Q000175:diagnosis* / Q000382:microbiology*; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D012680:Sensitivity and Specificity
D017931:DNA Primers