JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2014, Vol. 52 ›› Issue (10): 55-60.doi: 10.6040/j.issn.1671-7554.0.2014.426

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Detection of Mycoplasma pneumoniae in nasopharyngeal swab samples collected from children with acute respiratory infections by loop-mediated isothermal amplification

LI Dan, LI Jingyi, DONG Yanqing, REN Yi, TIAN Xiujun, XIN Deli   

  1. Beijing Friendship Hospital Affiliated to Capital Medical University;
    Beijing Tropical Medicine Research Institute;
    Beijing Key Laboratory for Research on Prevention and Treatment of Tropical Diseases, Beijing 100050, China
  • Received:2014-06-30 Revised:2014-09-17 Online:2014-10-10 Published:2014-10-10

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Abstract: Objective To establish a method of loop-mediated isothermal amplification(LAMP) for rapidly detecting Mycoplasma pneumonia (MP) in nasopharyngeal swab samples collected from children with acute respiratory infections. Methods According to the repeat sequence SDC1 of MP genome, 6 special primers for LAMP(SMP) were designed and the method for detecting MP DNA was developed. Sensitivity of the LAMP using SMP primers was tested using MP type strain FH DNA, and specificity was evaluated through cross-reaction with other Mycoplasmas and bacterial DNAs. The sensitivity of SMP was also compared with the reported LAMP primers(JMP) for MP. To test the reliability of SMP, 55 nasopharyngeal swab samples were detected by the LAMP using SMP primers and real-time PCR (Q-PCR), respectively. Results The LAMP using SMP primers could detect 6 copies of FH DNA, and no amplification was shown in DNA of other Mycoplasma or bacterials. The sensitivity of SMP was ten times higher than that of JMP. For 55 clinical specimens, the total consistency rate of LAMP methods using SMP and JMP primers, respectively, was 98.2%, and the total consistency rate of Q-PCR and LAMP using SMP primers was 96.4%. Conclusion LAMP method using the SMP primers designed in this research is more sensitive compared with the reported LAMP method for MP. Also, the LAMP method is more rapid and convenient for detecting MP from nasopharyngeal swab samples, so this method should be very useful for the rapid detection of MP in clinical diagnosis of MP infection.

Key words: Mycoplasma pneumoniae, Child, Respiratory tract infections, Loop-mediated isothermal amplification

CLC Number: 

  • R446.5
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