Real-time PCR for indication of representatives of Chlamydiales order: needs for development and main analytical characteristics of the method
https://doi.org/10.29235/1029-8940-2023-68-4-315-324
Abstract
Respiratory diseases occupy a dominant position in the structure of morbidity, often the etiology of which remains unclear. This situation is largely due to the lack of knowledge of the entire spectrum of pathogens that cause respiratory diseases, as well as the lack of approaches to their indication and identification. Thanks to constant progress of molecular biology, a number of bacteria related to chlamydia have been identified, some of which are associated with human respiratory diseases, including chlamydia-like microorganisms. The role of atypical pathogens such as chlamydia-like microorganisms Parachlamydia acanthamoebae, and Simkania negevensis in the pathology of the human respiratory tract is still not fully understood. A real-time PCR-based method has been developed and optimized. It is capable of detecting the DNA of both chlamydia and chlamydia-like microorganisms (Parachlamydia acanthamoebae and Simkania negevensis). The method is characterized by high specificity and reproducibility, as well as by analytical sensitivity at a level of 8.7∙103 GE/ml. Its use will make it possible to assess the contribution of these atypical pathogens to the etiological structure of infectious diseases of the respiratory tract.
About the Authors
Yu. M. KapustinaBelarus
Yulia M. Kapustina, Postgraduate student, Junior Researcher
220114
23, Filimonov Str.
Minsk
L. V. Rubanik
Belarus
Lyudmila V. Rubanik, Ph. D. (Biol.), Associate Professor, Head of the Laboratory
220114
23, Filimonov Str.
Minsk
N. P. Shmeleva
Belarus
Natalia P. Shmeleva, Ph. D. (Med.), Head of the Laboratory
220114
23, Filimonov Str.
Minsk
N. V. Sivets
Belarus
Natalia V. Sivets, Researcher
220114
23, Filimonov Str.
Minsk
E. E. Grigorieva
Belarus
Elena E. Grigorieva, Ph. D. (Biol.), Leading Researcher
220114
23, Filimonov Str.
Minsk
References
1. Ochilova S. S., Edgorova N. T., Ernaeva G. Kh. The role of Mycoplasma pneumonia as etiological agent in disease of respiration tracts. Biologiya i integrativnaya meditsina [Biology and Integrative medicine], 2017, no. 4, pp. 110–128 (in Russian).
2. Greub G. Parachlamydia acanthamoebae, an emerging agent of pneumonia. Clinical Microbiology and Infection, 2009, vol. 15, no. 1, pp. 18–28. doi: 10.1111/j.1469-0691.2008.02633.x
3. Kosyakov S. Ya., Angotoeva I. B. Acute respiratory infections in ENT practice. Meditsinskii sovet [Medical Council], 2013, no. 7, pp. 26–31 (in Russian).
4. Hokynar K., Kurkela S., Nieminen T., Saxen H., Vesterinen E. J., Mannonen L., Pietikäinen R., Puolakkainen M. Parachlamydia acanthamoebae detected during a pneumonia outbreak in southeastern Finland, in 2017–2018. Micro organisms, 2019, vol. 7, no. 5, p. 141. doi: 10.3390/microorganisms7050141
5. Lienard J., Croxatto A., Aeby S., Jaton K., Posfay-Barbe K., Gervaix A., Greub G. Development of a new chlamydiales-specific real-time PCR and its application to respiratory clinical samples. Journal of Clinical Microbiology, 2011, vol. 49, no. 7, pp. 2637–2642. doi: 10.1128/JCM.00114-11
6. Taylor-Brown A., Vaughan L., Greub G., Timms P., Polkinghorne A. Twenty years of research into Chlamydia-like organisms: a revolution in our understanding of the biology and pathogenicity of members of the phylum Chlamydiae. Pathogens and Disease, 2015, vol. 73, no. 1, pp. 1–15. doi: 10.1093/femspd/ftu009
7. Vouga M., Baud D., Greub G. Simkania negevensis, an insight into the biology and clinical importance of a novel member of the Chlamydiales order. Critical Reviews in Microbiology, 2017, vol. 43, no. 1, pp. 62–80. doi: 10.3109/1040841X.2016.1165650
8. Pillonel T., Bertelli C., Salamin N., Greub G. Taxogenomics of the order Chlamydiales. International Journal of Systematic and Evolutionary Microbiology, 2015, vol. 6, pp. 1381–1393. doi: 10.1099/ijs.0.000090
9. Al-Younes H. M., Al-Zereini W., Obeidat N. M. Molecular evidence for the absence of an association between Simkania negevensis and respiratory diseases. Journal of Medical Microbiology, 2017, vol. 66, no. 9, pp. 1324–1327. doi: 10.1099/jmm.0.000564
10. Greub G., Mege J.-L., Raoult D. Parachlamydia acanthamoeba enters and multiplies within human macrophages and induces their apoptosis. Infection and Immunity, 2003, vol. 71, no. 10, pp. 5979–5985. doi: 10.1128/IAI.71.10.5979-5985.2003
11. Corsaro D., Venditti D., Valassina V. New chlamydial lineages from freshwater samples. Microbiology, 2002, vol. 148, no. 2, pp. 343–344. doi: 10.1099/00221287-148-2-343
12. Casson N., Medico N., Bille J., Greub G. Parachlamydia acanthamoebae enters and multiplies within pneumocytes and lung fibroblasts. Microbes and Infection, 2006, vol. 8, pp. 1294–1300. doi: 10.1016/j.micinf.2005.12.011
13. Pilloux L., Casson N., Sommer K., Klos A., Stehle J. C., Pusztaszeri M., Greub G. Severe pneumonia due to Parachlamydia acanthamoebae following intranasal inoculation: A mice model. Microbes and Infection, 2015, vol. 17, no. 11–12, pp. 755–760. doi: 10.1016/j.micinf.2015.08.007
14. Greub G., Hartung O., Adekambi T., Alimi Y. S., Raoult D. Chlamydialike organisms and atherosclerosis. Emerging Infectious Diseases, 2006, vol. 12, no. 4, pp. 705–706. doi: 10.3201/eid1204.050751
15. Kahane S., Greenberg D., Friedman M. G., Haikin H., Dagan R. High prevalence of “Simkania Z”, a novel Chlamydia-like bacterium, in infants with acute bronchiolitis. Journal of Infectious Diseases, 1998, vol. 177, no. 5, pp. 1425–1429. doi: 10.1086/517830
16. Kahane S., Greenberg D., Newman N., Dvoskin B., Friedman M. G. Domestic water supplies as a possible source of infection with Simkania. Journal of Infection, 2007, vol. 54, no. 1, pp. 75–81. doi: 10.1016/j.jinf.2006.01.011
17. Greenberg D., Banerji A., Friedman M. G., Chiu C. H., Kahane S. High rate of Simkania negevensis among Canadian inuit infants hospitalized with lower respiratory tract infections. Scandinavian Journal of Infectious Diseases, 2003, vol. 35, pp. 506–508. doi: 10.1080/00365540310014648
18. Kumar S., Kohlhoff S. A., Gelling M., Roblin P. M., Kutlin A., Kahane S., Friedman M. G., Hammerschlag M. R. Infection with Simkania negevensis in Brooklyn, New York. Pediatric Infectious Disease Journal, 2005, vol. 24, no. 11, pp. 989–992. doi: 10.1097/01.inf.0000183755.24578.0b
19. Friedman M. G., Kahane S., Dvoskin B., Hartley J. W. Detection of Simkania negevensis by culture, PCR, and serology in respiratory tract infection in Cornwall, UK. Journal of Clinical Pathology, 2006, vol. 59, no. 3, pp. 331–333. doi: 10.1136/jcp.2004.025601
20. Casson N., Posfay-Barbe K. M., Gervaix A., Greub G. New diagnostic real-time PCR for specific detection of Parachlamydia acanthamoebae DNA in clinical samples. Journal of Clinical Microbiology, 2008, vol. 46, no. 4, pp. 1491–1493. doi: 10.1128/JCM.02302-07
21. Lamoth F., Aeby S., Schneider A., Jaton-Ogay K., Vaudaux B., Greub G. Parachlamydia and Rhabdochlamydia in premature neonates. Emerging Infectious Diseases, 2009, vol. 15, no. 12, pp. 2072–2075. doi: 10.3201/eid1512.090267
22. Kose M., Ekinci D., Gokahmetoglu S., Elmas T., Öztürk M. K. Simkania negevensis: Is it a real respiratory pathogen? Acta Microbiologica et Immunologica Hungarica, 2015, vol. 62, no. 2, pp. 161–166. doi: 10.1556/030.62.2015.2.6
23. Niemi S., Greub G., Puolakkainen M. Chlamydia-related bacteria in respiratory samples in Finland. Microbes and Infection, 2011, vol. 13, no. 10, pp. 824–827. doi: 10.1016/j.micinf.2011.04.012
24. Fomina E. G., Rubanik L. V., Kapustina Yu. M., Grigor’eva E. E., Zverko V. V. Construction of recombinant plasmids containing diagnostically significant regions of the Chlamydia trachomatis genome. Sovremennye problemy infektsionnoi patologii cheloveka: sbornik nauchnykh trudov [Modern problems of human infectious pathology: a collection of scientific papers]. Minsk, 2020, vol. 13, pp. 104–109 (in Russian).
25. Lorenz T. C. Polymerase chain reaction: basic protocol plus troubleshooting and optimization strategies. Journal of Visualized Experiments, 2012, vol. 22, no. 63, p. e3998. doi: 10.3791/3998
26. Vodchits N. V., Yurchenko E. O., Volotovich A. A. Study of the optimal conditions for ISSR–PCR in marking of Vaccinium corymbosum cultivars. Vesnіk Paleskaga dzyarzhaunaga unіversіteta. Seryya pryrodaznauchykh navuk [Bulletin of Polessky State University. Natural Sciences Series], 2015, no. 1, pp. 3–7 (in Russian).