Journal of Reproduction & Infertility

Journal of Reproduction & Infertility

Detection of Legionella pneumophila by nested PCR-RFLP and ELISA on urine specimens of pregnant women with respiratory infections

Authors
Leili Chamani Tabriz 1
Mahmood Jeddi-Tehrani 2
Shila Motahari 3
Hojjat Zeraati 4
Soheila Asgari 1
Mohammad Karegar 3
Mahyar Ostad Karampour 2
1 Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
2 Monoclonal Antibody Research Center, Avicenna Research Institute (ACECR), Tehran, Iran
3 Department of Microbiology, Azad University of Jahrom, Jahrom, Iran
4 Epidemiology Department, Faculty of Publi Health, Tehran Medical Sciences University, Tehran, Iran
Abstract
Introduction: Pneumonia during pregnancy can induce serious consequences to the mother and the fetus, therefore its diagnosis and therapy is very important. There are few published jri_articles on Legionella infection prevalence during pregnancy. In patients with Legionellosis, bacterial LPS and DNA are excreted into urine for extended periods, so combination of PCR and ELISA methods would be a good diagnostic tool. This research was done to determine the prevalence of L. pneumophila in pregnant women with respiratory infections. Materials & Methods: This is a cross-sectional study on 95 pregnant women with respiratory infection carried out during winter to summer 2006. Presence of Legionella infection was con-firmed by nested PCR-RFLP and antigen detection in urine specimens by ELISA method. The data were analyzed by SPSS, version 13, by using independent t tests, Fisher’s exact test, χ2, a logistic model and McNemar’s test, while considering p < 0.05 as significant. Results: The prevalence of infection using PCR was 22.1% (CI=14.1%-30.1%) and by ELISA it was 4.2% (CI=2%-8.2%); this difference was statistically significant (p < 0.005). The most pre-valent clinical features were Cough (56.8%), headache (54.7%), abdominal pain (38.9%), chills (35.8%), fever (22.1%) and diarrhea (8.4%). There were significant statistical relationships bet-ween cases with a positive CRP and fever, chills and abdominal pain and previous liver or renal problems (p < 0.05, p < 0.001). There were significant relationships between fever and chills with ELISA results (p < 0.05) but no relationships with other variables. Conclusion: There was a considerable prevalence of this infection in the studied population (22.1%). It seems that performing PCR & ELISA tests on urine sample is suitable in detecting Legionella species and it can provide results in a less than a day_ a great help in the diagnosis and treatment of pneumonia especially during pregnancy.
Keywords
ELISA
Legionella pneumophila
molecular
Pneumonia
Pregnancy
  1. Sohn J W, Park S C, Choi Y H, Woo H J, Cho Y K, Lee J S, et al. Atypical pathogens as etiologic agents in hospitalized patients with community-acquired pneu-monia in Korea: a prospective multi-center study. J Korean Med Sci. 2006;21(4):602-7.
  2. Ngeow Y F, Suwanjutha S, Chantarojanasriri T, Wang F, Saniel M, Alejandria M, et al. An Asian study on the prevalence of atypical respiratory pathogens in community-acquired pneumonia. Int J Infect Dis. 2005; 9(3):144-53.
  3. Behbehani N, Mahmood A, Mokaddas E M, Bittar Z, Jayakrishnan B, Khadadah M, et al. Significance of atypical pathogens among community-acquired pneu-monia adult patients admitted to hospital in Kuwait. Med Princ Pract. 2005;14(4):235-40.
  4. Nagai T, Sobajima H, Iwasa M, Tsuzuki T, Kura F, Amemura-Maekawa J, et al. Neonatal sudden death due to legionella pneumophila associated with water birth in a domestic spa bath, J Clin Microbiol. 2003; 41(5):2227-9.
  5. Montagna M T, Napoli C, Tato D, Spilotros G, Barbuti G, Barbuti S. Clinical-environmental surveil-lance of legionellosis: an experience in southern Italy. Eur J Epidemiol. 2006;21(4):325-31.
  6. Murdoch D R, Walford E J, Jennings L C, Light G j, Schouseboe M I, Chershsky AY, et al. Use the poly-merase chain reaction to detect legionella DNA in urine and serum samples from patient with pneumonia. Clin Infect Dis. 1997;25(4):939.
  7. Blazquez R M, Espinosa F J, Martinez-Toldos C M, Alemany L, Garcia-Orenes M C, Segovia M. Sensiti- vity of urinary antigen test in relation to clinical severity in a large outbreak of Legionella pneumonia in Spain. Eur J Clin Microbiol Infect Dis. 2005;24(7): 488-91.
  8. KO K S, Hong S K, Lee K H, Lee H K, Park M Y, Miyamoto H, et al. Detection and identification of legionella pneumophila by PCR-restriction fragment length polymorphism analysis of the RNA polymerase gene (rpoB). J Microbial Methods. 2003; 54:325-337.
  9. Murdoch D R. Diagnosis of legionella infection. Clin Infect Dis. 2003;36(1):64-9
  10. Edelstein P H, Cianciotto N P. Legionella, Mandell G L, Dolin R, Bennette J E. 6th Editin, Published by Else-vier Churchill Livingstone, Philadelphia. 2005;p: 2239-51.
  11. Fields B S, Benson R F, Besset R E. Legionella and Legionnaires' disease: 25 years of investigation. Clin Microbiol Rev. 2002;15(3):506-26.
  12. Kim M J, Sohn J W, Park D W, Park S C, Chun B C. Characterization of a lipoprotein common to Legio-nella species as a urinary broad-spectrum antigen for diagnosis of Legionnaires' disease. J Clin Microbiol. 2003;41(7):2974-9.
  13. Eisenberg V H, Eidelman L A, Arbel R, Ezra Y. Legionnaire's disease during pregnancy: a case presen-tation and review of the literature. Eur J Obstet Gynecol Reprod Biol. 1997;72(1):15-8.
  14. Vimercati A, Greco P, Bettocchi S, Resta L, Selvaggi L. Legionnaire's disease complicating pregnancy: a case report with intrauterine fetal demise. J Perinat Med. 2000;28(2):147-50.
  15. Sambrook J, Russell DW. Preparation and analysis of eukaryotic genomic DNA, Molecular Cloning: A Laboratory Manual. 3rd Edition. Published by Cold Spring Harbor Laboratory Press, 2001;pp:6.28-6.30.
  16. Nielsen K, Hindersson P, Hoiby N, Bangsborg J M. Sequencing of the rpoB Gene in Legionella pneu-mophila and Characterization of Mutations Associated with Rifampin Resistance in the Legionellaceae. Anti-microb Agents Chemother. 2000;44(10):2679-2683.
  17. Sopena N, Pedro-botet M L, Sabria M, Garsia D. Comparative study of community-acquired pneumonia caused by streptococcus pneumoniae, legionella pneu-mophila or chlamydia pneumoniae. Scand J Infect Disease. 2004;36:330-334.
  18. Welte T, Marre R, Suttorp N. What is new in the treatment of community-acquired pneumonia. Med Klin. 2006;101(4):313-20.
  19. Huang H H, Zhang Y Y, Xiu Q Y, Zhou X, Huang S G, Lu Q, et al. Community-acquired pneumonia in Shanghai, China: microbial etiology and implications for empirical therapy in a prospective study of 389 patients. Eur J Clin Microbiol Infect Dis. 2006;25(6): 369-74.
  20. Al-Ali M K, Batchoun R G, Al-Nour T M. Etiology of community-acquired pneumonia in hospitalized patients in Jordan. Saudi Med J. 2006; 27(6):813-6.
  21. Gaillac N, Floccard B, Ould T, Benatir F, Levrat A, Meunier P, et al. Legionella pneumophila pneumonia during pregnancy: a case report, J Infect. 2006;52(6): 63-4.
  22. Su H P, Tseng L R, Chou C Y, Chung T C, Pan T M. Legionella pneumophila infection in the Taiwan area. J Infect Chemother. 2005;11(5):244-9.
  23. حسینی دوست سید رضا، سیل دیوید. جداسازی لژیونلا از سیستم آب بیمارستانها (بخشهای پیوند عضو). مجله پزشکی کوثر. سال سوم(1377)، شماره سوم، صفحات150-145.
  24. خسرو شاهی نادر، موسویان مجتبی. جداسازی و تعیین هویت عوامل بیماری لژیونر از تجهیزات درمانی و منابع محیطی، مجله دانشگاه علوم پزشکی و خدمات بهداشتی- درمانی قزوین. 1382، شماره 29، صفحات 74-70.
Journal of Reproduction & Infertility
Volume 8, Issue 3
Autumn 2007
Pages 221-229