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Advanced and conventional molecular techniques in the diagnosis of mycoplasma pneumoniae in patients with respiratory tract infection
Joint Authors
Muhammad, Umar A.
Hasan, Muntha M.
al-Uqayli, Mushtaq T. S.
Muhammad, Nidal A.
Source
Journal of the Faculty of Medicine Baghdad
Issue
Vol. 56, Issue 1 (31 Mar. 2014), pp.95-100, 6 p.
Publisher
University of Baghdad Faculty of Medicine
Publication Date
2014-03-31
Country of Publication
Iraq
No. of Pages
6
Main Subjects
Topics
Abstract EN
Background : M.
pneumoniae is an important human pathogen that produces community-acquired respiratory tract infection.
Diagnosis of M.
pneumoniae infection is challenging and crucial for the timely initiation of the effective antibiotic therapy.
Objective : This study has been undertaken to detect M.
pneumoniae in respiratory samples (throat swabs, throat wash and sputum) in patients with respiratory tract infection qualitatively by conventional polymerase chain reaction (PCR).
Also, more advanced one, real time PCR was used to determine mycoplasmal target gene qualitatively and quantitatively.
Patients and methods : The study was performed on Seventy-five patients and thirty healthy subject as control.
Genomic DNA was extracted and M.
pneumoniae target gene (lipoprotein gene) was amplified using conventional PCR.
Negative, positive controls and internal controls were involved in each experimental run.
The amplified products were analyzed in 2 % agarose gel and visualized using Red safe staining.
In real time PCR, specific primer and probe mix depending on TaqMan® principle was used to detect PI adhesion gene through FAM channel.
A fluorogenic probe was included in the same reaction mixture which consists of a DNA probe labeled with a 5'-dye and a 3'-quencher.
During PCR amplification Data were analyzed using Smart-cycler software and M.
pneumoniae DNA copy number was estimated from the cross point threshold relative to positive standard.
Results : Thirty five patients(45.5 %) were positive by PCR and Thirty two (42.6 %) were positive by Realtime PCR.
The highest rate of infection by using tow molecular methods were of less than 20 years of age.
The quantity of M.
pneumoniae DNA target gene in positive Real-time PCR were ranged between 10-2000copiesxl.
Conclusion : The study concluded that both of molecular techniques conventional and real-time PCR are a rapid, reliable and ideal in diagnosis of M.
pneumoniae using throat swabs, throat wash and sputum samples.
American Psychological Association (APA)
Hasan, Muntha M.& al-Uqayli, Mushtaq T. S.& Muhammad, Nidal A.& Muhammad, Umar A.. 2014. Advanced and conventional molecular techniques in the diagnosis of mycoplasma pneumoniae in patients with respiratory tract infection. Journal of the Faculty of Medicine Baghdad،Vol. 56, no. 1, pp.95-100.
https://search.emarefa.net/detail/BIM-385006
Modern Language Association (MLA)
Hasan, Muntha M.…[et al.]. Advanced and conventional molecular techniques in the diagnosis of mycoplasma pneumoniae in patients with respiratory tract infection. Journal of the Faculty of Medicine Baghdad Vol. 56, no. 1 (2014), pp.95-100.
https://search.emarefa.net/detail/BIM-385006
American Medical Association (AMA)
Hasan, Muntha M.& al-Uqayli, Mushtaq T. S.& Muhammad, Nidal A.& Muhammad, Umar A.. Advanced and conventional molecular techniques in the diagnosis of mycoplasma pneumoniae in patients with respiratory tract infection. Journal of the Faculty of Medicine Baghdad. 2014. Vol. 56, no. 1, pp.95-100.
https://search.emarefa.net/detail/BIM-385006
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references : p. 99-100
Record ID
BIM-385006