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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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Original Research Articles                      Volume : 13, Issue:6, June, 2024

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com /
submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2024.13(6): 24-31
DOI: https://doi.org/10.20546/ijcmas.2024.1306.002


Direct Detection of Atypical Bacteria in Clinical Samples from Patients with Chronic Obstructive Pulmonary Disease (COPD) by PCR and RT-PCR
Department of Microbiology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi- 110007, India
*Corresponding author
Abstract:

Chronic obstructive pulmonary disease (COPD) represents an important public health challenge and is a major cause of morbidity and mortality throughout the world. The chronic course of COPD is often accompanied by acute exacerbations of COPD (AECOPD) where 70% of exacerbations are caused by infection due to aerobic bacteria, 30% due to viruses and 5-10 % by atypical bacteria, most commonly Mycoplasma pneumoniae (M. pneumoniae), Chlamydia pneumoniae (C. pneumoniae) and Legionella pneumophila (L. pneumophila). Identification of these atypical microorganism is challenging, however, with the availability of newer molecular diagnostic techniques such as PCR and Real time PCR (RT-PCR) the sensitivity in detecting these pathogens has improved and can be tested simultaneously. So, the present prospective study was undertaken to determine the atypical pathogens in patients with COPD by PCR and RT-PCR in the clinical samples. 196 hospitalized patients were included in the study. They were categorized into different stages of COPD and AECOPD using GOLD and Anthonisen criteria. Clinical samples such as throat swab, nasopharyngeal swab, sputum, were collected. Atypical bacteria were identified by using PCR and RT-PCR methods. All these patients showed varying degree of exacerbation. Majority, 78/196 (39.79%) of the patients belonged to severe COPD category. Whereas, 96/193(49.74%) patients had only mild exacerbation. PCR was found to be positive in 11 and 22 numbers for M. pneumoniae and L.pneumophila. None of the samples were positive for Chlamydia pneumophila. Presence of pathogens, atypical bacteria did not show any correlation with the staging of AECOPD. The results were not statistically significant. No one method is suitable for the detection of atypical bacteria. Hence, a combination of tests for identification of Atypical pathogens is required for better sensitivity.


Keywords: Atypical bacteria, COPD, AECOPD, PCR, RT-PCR


References:

Anthonisen NR, Manfreda J, Warren CP, Hershfield ES, Harding GK and Nelson NA. Antibiotic therapy in exacerbations of chronic obstructive pulmonary disease. Ann Intern Med 1987;106:196–204.

Beaty C D, Grayston J T, Wang S P, Kuo C C, Reto C S, Martin T R. Chlamydia pneumoniae, strain TWAR, infection in patients with chronic obstructive pulmonary disease. Am Rev Respir Dis 1991; 144: 1408–1410. https://doi.org/10.1164/ajrccm/144.6.1408

Criner, G. J., Bourbeau, J., Diekemper, R. L., Ouellette, D. R., Goodridge, D., Hernandez, P., et al., (2015). Prevention of acute exacerbations of COPD: American College of Chest Physicians and Canadian Thoracic Society Guideline. Chest 147, 894–942 https://doi.org/10.1378/chest.14-1676

Diederen B M W, Valk P D L P M van der, Kluytmans J A. W J, Peeters M F, Hendrix R. The role of atypical respiratory pathogens in exacerbations of chronic obstructive pulmonary disease. European Respiratory Journal (Internet). 2007 Aug 1 (cited 2022 Feb 8);30(2):240–4. https://doi.org/10.1183/09031936.00012707

Ewig S. Legionella spp. in acute exacerbations of chronic obstructive pulmonary disease: what is the evidence? Eur Respir J 2002; 19: 387–389. https://doi.org/10.1183/09031936.02.00281402

Global initiative for chronic obstructive lung disease (GOLD) 2018.

Global Initiative for Chronic Obstructive Lung Disease. (2021). Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease. 2022 report (Online).  

Gnarpe, J., A. Lunbäck, H. Gnarpe, and B. Sundelöf. 1997. Comparison of nasopharyngeal and throat swabs for the detection of Chlamydia pneumoniae and Mycoplasma pneumoniae by polymerase chain reaction. Scand. J. Infect. Dis. Suppl. 104:11-12.

Gullsby K, Storm M, Bondeson K. Simultaneous detection of Chlamydophila pneumoniae and Mycoplasma pneumoniae by use of molecular beacons in a duplex real-time PCR. J Clin Microbiol 2008; 46: 727–731. https://doi.org/10.1128/JCM.01540-07

Hahn D L. Chlamydia pneumoniae, asthma, and COPD: what is the evidence?Ann Allergy Asthma Immunol 1999; 83: 271–288. https://doi.org/10.1016/S1081-1206(10)62666-X

Honda, J., T. Yano, M. Kusaba, J. Yonemitsu, H. Kitajima, M. Masuoka, K. Hamada, and K. Oizumi. 2000. Clinical use of capillary PCR to diagnose Mycoplasma pneumoniae. J. Clin. Microbiol. 38:1382-1384. https://doi.org/10.1128/jcm.38.4.1382-1384.2000  

Lieberman D, Ben-Yaakov M, Lazarovich Z, Ohana B, Boldur I. Chlamydia pneumoniae infection in acute exacerbations of chronic obstructive pulmonary disease: analysis of 250 hospitalizations. Eur J Clin Microbiol Infect Dis 2001; 20: 698–704. https://doi.org/10.1007/s100960100596.

Lieberman D, Dvoskin B, Lieberman D V, Kahane S, Friedman M G. Serological evidence of acute infection with the Chlamydia-like microorganism Simkania negevensis (Z) in acute exacerbation of chronic obstructive pulmonary disease. Eur J Clin Microbiol Infect Dis 2002; 21: 307–309. https://doi.org/10.1007/s10096-002-0703-7

Lieberman D, Lieberman D, Ben-Yaakov M, et al., Infectious etiologies in acute exacerbation of COPD. Diagn Microbiol Infect Dis 2001; 40: 95–102. https://doi.org/10.1016/s0732-8893(01)00255-3

Lieberman D, Lieberman D, Ben-Yaakov M, et al., Serological evidence of Mycoplasma pneumoniae infection in acute exacerbation of COPD. Diagn Microbiol Infect Dis 2002; 44: 1–6. https://doi.org/10.1016/s0732-8893(02)00421-2

Lieberman D, Lieberman D, Shmarkov O, et al., Serological evidence of Legionella species infection in acute exacerbation of COPD. Eur Respir J 2002; 19: 392–397. https://doi.org/10.1183/09031936.02.00256702

Murdoch D R. Molecular genetic methods in the diagnosis of lower respiratory tract infections. APMIS 2004; 112: 713–727. https://doi.org/10.1111/j.1600-0463.2004.apm11211-1202.x

Murdoch, D R., Diagnosis of Legionella infection. Clin Infect Dis 2003; 36: 64–69. https://doi.org/10.1086/345529

Schoonbroodt S, Ichanté J L, Boffé S, Devos N, Devaster J M, Taddei L, Rondini S, Arora A K, Pascal T, Malvaux L. Real-time PCR has advantages over culture-based methods in identifying major airway bacterial pathogens in chronic obstructive pulmonary disease: Results from three clinical studies in Europe and North America. Front. Microbiol. 2023;13:1098133. https://doi.org/10.3389/fmicb.2022.1098133

Sethi S. Infectious etiology of acute exacerbations of chronic bronchitis. Chest 2000;117:380. https://doi.org/10.1378/chest.117.5_suppl_2.380s

Welti M, Jaton K, Altwegg M, Sahli R, Wenger A, Bille J. Development of a multiplex real-time quantitative PCR assay to detect Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoniae in respiratory tract secretions. Diagn Microbiol Infect Dis 2003; 45: 85–95. https://doi.org/10.1016/s0732-8893(02)00484-4

Williamson J, Marmion BP, Worswick D A, Kok T W, Tannock G, Herd R, Harris R J, 1992. Laboratory diagnosis of Mycoplasma pneumoniae infection, antigen capture and PCR-gene amplification for detection of the Mycoplasma, problems of clinical correlation. Epidemiol Infect 1992; 109: 519–537 https://doi.org/10.1017/s0950268800050512

World Health Organization. Global Surveillance, Prevention and Control of Chronic Respiratory Diseases: A Comprehensive Approach. Geneva, Switzerland: World Health Organization; 2007.


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How to cite this article:

Tonushyam Sonowal and Malini Shariff. 2024. Direct Detection of Atypical Bacteria in Clinical Samples from Patients with Chronic Obstructive Pulmonary Disease (COPD) by PCR and RT-PCR.Int.J.Curr.Microbiol.App.Sci. 13(6): 24-31. doi: https://doi.org/10.20546/ijcmas.2024.1306.002
Copyright: This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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