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
[1] Waites K B, Balish M F, Atkinson T P. New insights into the pathogenesis and detection of Mycoplasma pneumoniae infections[J]. Future Microbiol, 2008, 3(6): 635-648.
[2] Ieven M, Ursi D, Van Bever H, et al. Detection of Mycoplasma pneumoniae by two polymerase chain reactions and role of M. pneumoniae in acute respiratory tract infections in pediatric patients[J]. J Infect Dis, 1996, 173(6): 1445-1452.
[3] Welti M, Jaton K, Altwegg M, et al. Development of a multiplex real-time quantitative PCR assay to detect Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae in respiratory tract secretions[J]. Diagn Microbiol Infect Dis, 2003, 45(2): 85-95.
[4] Sánchez-Vargas F M, Gómez-Duarte O G. Mycoplasma pneumoniae-an emerging extra-pulmonary pathogen[J]. Clin Microbiol Infect, 2008, 14(2): 105-117.
[5] Ni X, McManus D P, Yan H, et al. Loop-Mediated Isothermal Amplification (LAMP) assay for the identification of Echinococcus multilocularis infections in canine definitive hosts[J]. Parasit Vectors, 2014, 7(1): 254.
[6] Fallahi S, Seyyed Tabaei S J, Pournia Y, et al. Comparison of loop-mediated isothermal amplification (LAMP) and nested-PCR assay targeting the RE and B1 gene for detection of Toxoplasma gondii in blood samples of children with leukaemia[J]. Diagn Microbiol Infect Dis, 2014, 79(3): 347-354.
[7] Dhama K, Karthik K, Chakraborty S, et al. Loop-mediated isothermal amplification of DNA (LAMP): a new diagnostic tool lights the world of diagnosis of animal and human pathogens: a review[J]. Pak J Biol Sci, 2014, 17(2): 151-166.
[8] Zhou D, Guo J, Xu L, et al. Establishment and application of a loop-mediated isothermal amplification (LAMP) system for detection of cry1Ac transgenic sugarcane[J]. Sci Rep, 2014, 4: 4912.
[9] 何旭峰, 彭焕, 丁中, 等. 植物罹病组织中象耳豆根结线虫的LAMP快速检测方法[J]. 中国农业科学, 2013, 46(3):534-544. HE Xufeng, PENG Huan, DING Zhong, et al. Loop-mediated isothermal amplification assay for rapid diagnosis of meloidogyne enterolobii directly from infected plants[J]. Scientia Agricultura Sinica, 2013, 46(3):534-544.
[10] Gotoh K, Nishimura N, Ohshima Y, et al. Detection of Mycoplasma pneumoniae by loop-mediated isothermal amplification (LAMP) assay and serology in pediatric community-acquired pneumonia[J]. J Infect Chemother, 2012, 18(5): 662-667.
[11] 侯安存, 刘玉华, 辛德莉, 等. 健康儿童鼻咽部常见致病微生物携带状况及临床意义[J]. 中华儿科杂志, 2002, 40(1):45-49. HOU Ancun, LIU Yuhua, XIN Deli, et al. Nasopharyngeal carriage status of common pathogenic microorganisms in healthy children and its clinical significance[J]. Chin J Pediatr, 2002, 40(1): 45-49.
[12] 胡亚美, 江载芳, 诸福棠. 实用儿科学[M]. 7版.北京:人民卫生出版社, 2002: 1204-1205.
[13] 黄庆华, 叶嗣颖, 陈桂蓝. 不同临床样品肺炎支原体的分离培养[J]. 临床检验杂志, 1998, 16(04): 51-52.
[14] 赵茹, 黄莉, 季伟. 双份血清MP抗体及痰MP-PCR检测在儿童肺炎支原体感染诊断中的价值[J]. 江苏医药, 2008, 34(12): 1282-1283.
[15] Welti M, Jaton K, Altwegg M, et al. Development of a multiplex real-time quantitative PCR assay to detect Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae in respiratory tract secretions[J]. Diagn Microbiol Infect Dis, 2003, 45(2): 85-95.
[16] Kenri T, Taniguchi R, Sasaki Y, et al. Identification of a new variable sequence in the P1 cytadhesin gene of Mycoplasma pneumoniae: evidence for the generation of antigenic variation by DNA recombination between repetitive sequences[J]. Infect Immun, 1999, 67(9): 4557-4562.
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