Rapid Detection of Mycoplasma pneumoniae by Loop-Mediated Isothermal Amplification (LAMP) in Clinical Respiratory Specimens

  • Maryam ARFAATABAR Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Narjes NOORI GOODARZI 1. Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Davoud AFSHAR Department of Microbiology and Virology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran
  • Hamed MEMARIANI Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran
  • Ghasem AZIMI Department of Internal Medicine, Shahed University of Medical Sciences, Tehran, Iran
  • Ensieh MASOORIAN Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Reza POURMAND 1. Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2. Biotechnology Research Center, Tehran University of Medical Sciences, Tehran, Iran
Keywords: Mycoplasma pneumoniae; PCR; Loop-mediated isothermal amplification (LAMP); Culture

Abstract

Background: Mycoplasma pneumoniae is a common cause of community-acquired pneumonia (CAP) worldwide, especially among children and debilitated populations. The present study aimed to investigate a loop-mediated isothermal amplification (LAMP) technique for rapid detection of M. pneumoniae in clinical specimens collected from patients with pneumonia. Methods: Throat swabs were collected from 110 outpatients who suffered from pneumonia. Throat swab samples were obtained from patients referred to the hospital outpatient clinics of Tehran University hospitals, Iran in 2017. The presence of M. pneumoniae in the clinical specimens was evaluated by LAMP, PCR and culture methods. Sensitivity and specificity of the LAMP and PCR assays were also determined. Results: Out of 110 specimens, LAMP assay detected M. pneumoniae in 35 specimens. Detection limit of the LAMP assay was determined to be 33fg /µL or ~ 40 genome copies/reaction. Moreover, no cross-reaction with genomic DNA from other bacteria was observed. Only 25 specimens were positive by the culture method. The congruence between LAMP assay and culture method was ‘substantial’ (κ=0.77). Specificity and sensitivity of LAMP assay were 88.2%, 100% in compare with culture. However, the congruence between LAMP assay and PCR assay was ‘almost perfect’ (κ=0.86). Specificity and sensitivity of LAMP assay were 92.5%, 100% in compare with PCR. Conclusion: Overall, the LAMP assay is a rapid and cost-efficient laboratory test in comparison to other methods including PCR and culture. Therefore, the LAMP method can be applied in identification of M. pneumoniae isolates in respiratory specimens.    

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Published
2019-05-15
How to Cite
1.
ARFAATABAR M, NOORI GOODARZI N, AFSHAR D, MEMARIANI H, AZIMI G, MASOORIAN E, POURMAND MR. Rapid Detection of Mycoplasma pneumoniae by Loop-Mediated Isothermal Amplification (LAMP) in Clinical Respiratory Specimens. Iran J Public Health. 48(5):917-924.
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Original Article(s)