Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
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Advances in Clinical and Experimental Medicine

2009, vol. 18, nr 4, July-August, p. 337–344

Publication type: original article

Language: English

Comparison of the Detectability of Mycoplasma pneumoniae Infection in Children Using PCR and Serological Methods: Indirect Immunofluorescence and Enzyme Immunnoassays

Porównanie wykrywalności zakażeń Mycoplasma pneumoniae u dzieci z użyciem metody PCR oraz metod serologicznych: immunofluorescencji pośredniej i odczynów immunoenzymatycznych

Beata Mączyńska1,, Marzenna Bartoszewicz1,, Urszula Kasprzykowska1,, Barbara Sozańska2,, Adam Junka1,

1 Department of Microbiology, Wrocław Medical University, Poland

2 Department of Pediatrics, Allergology and Cardiology, Wrocław Medical University, Poland

Abstract

Background. Mycoplasma pneumoniae is an important etiological agent of upper and lower respiratory tract infections, mainly atypical pneumonia, in children and young adults. A specific diagnosis is important because the treatment of M. pneumoniae infections with β−lactam antibiotics is ineffective, whereas macrolides or tetracyclines may markedly reduce the duration of the illness.
Objectives. Determination of the specificity and sensitivity of routine diagnostic procedures of M. pneumoniae using PCR and serological methods.
Material and Methods. Throat swabs and sera from children in the acute phase of atypical pneumonia were analyzed. The PCR assay was performed with the Venor Mp Diagnostic Kit (Minerva, Germany). The levels of IgM and IgG antibodies were evaluated in serum samples using ELISA from DiaSorin, USA, ELISA from Euroimmun, Germany, and an indirect immunofluorescence test with the BIOCHIP technique from Euroimmun.
Results. Fifty−two percent of the throat swabs were positive by PCR. Depending on the serological test used, different percentages of positive results (IgM or both IgM and IgG antibodies) were detected in the serum samples. In 15% of the cases, positive results with PCR were negative in all the serological methods.
Conclusion. The indirect fluorescence test was considered the most reliable and sensitive in comparison with PCR assay.

Streszczenie

Wprowadzenie. Mycoplasma pneumoniae jest ważnym etiologicznym czynnikiem zakażeń górnych i dolnych dróg oddechowych, głównie atypowego zapalenia płuc, u dzieci oraz młodych dorosłych. Postawienie trafnej diagnozy jest niezwykle istotne, ponieważ leczenie zakażeń Mycoplasma pneumoniae antybiotykami β−laktamowymi nie przynosi rezultatów, kuracja makrolidami lub tetracyklinami natomiast znacząco skraca czas trwania choroby.
Cel pracy. Oszacowanie swoistości i czułości procedur diagnostycznych stosowanych w diagnostyce zakażeń M. pneumoniae – metody PCR oraz metod serologicznych.
Materiał i metody. Pobierano wymazy gardła oraz surowicę od dzieci, w ostrej fazie infekcji zdiagnozowanej jako atypowe zapalenie płuc. Technikę PCR wykonano z użyciem Diagnostic Kit Venor Mp (Minerva, Germany). Stężenia przeciwciał IgG oraz IgM z próbek surowicy oznaczono z użyciem testu ELISA DiaSorin, USA, ELISA Euroimmun, Germany, test immunofluorescencji pośredniej przeprowadzono używając zestawu Biochip technique (Euroimmun, Germany).
Wyniki. Dla 52% przypadków wymazów z gardła zbadanych techniką PCR uzyskano wynik pozytywny. W zależności od rodzaju użytego testu serologicznego, uzyskano różny odsetek wyników dodatnich (dla przeciwciał IgM lub przeciwciał IgM oraz IgG) w zbadanych próbkach osocza. Dla 15% przypadków uzyskano pozytywne wyniki testu PCR i negatywne wyniki z użyciem wszystkich metod serologicznych.
Wnioski. Test immunofluorescencji pośredniej okazał się najbardziej wiarygodny i czuły w porównaniu z techniką PCR.

Key words

Mycoplasma pneumoniae, PCR, serological diagnosis

Słowa kluczowe

Mycoplasma pneumoniae, PCR, diagnostyka serologiczna

References (28)

  1. Mac Farlane J: An overview of community−acquired pneumonia with lessons learned from the British Thoracic Society Study. Semin Respir Infect 1994, 9, 153–165.
  2. Grendrel D, Raymond J, Moulin F, Inigues JL, Ravilly S, Habb F, Lebon P, Kalifa G: Etiology and response to antibiotic therapy of community−acquired pneumonia in French children. Eur J Clin Microbiol Infect Dis 1997, 16, 388–391.
  3. Principi N, Esposito S, Blasi F, Allegra L: Role of Mycoplasma pneumonia in children with communityacquired lower respiratory tract infections. Clin Infect Dis 2001, 32, 1281–1289.
  4. Kałużewski S, Jagielski M, Rastawicki W, Kochman M: The estimation of appearance of Mycoplasma pneumoniae infections in Poland 1970–1993 years, on the basis of serological study (in Polish). Przeg Epid 1994, 48, 166–175.
  5. Waris ME, Toikka P, Nikkari S, Meurman O, Vainionpaa R, Mertsol J, Ruuskanen O: Diagnosis of Mycoplasma pneumoniae pneumonia in children. J Clin Microbiol 1998, 36, 3155–3159.
  6. Mączyńska B, Stankiewicz M: Detection of Mycoplasma pneumoniae antibodies in patients with respiratory tract infections. Clin Microbiol Infect 2000, 6, (Suppl. 1), abstr TuP81: 55.
  7. Foy HM: Infections caused by Mycoplasma pneumoniae and possible carrier state in different populations of patients. Clin Infect Dis 1993, 17 (1), 37–47.
  8. Kenny RT, Li JS, Clyde WA, Wall TC, O’Connor CM, Campbell PT, Van Trigt P, Corey GR: Mycoplasmal pericarditis: evidence of invasive disease. Clin Infect Dis 1993, 17, 558–562.
  9. Rastawicki W, Jagielski M: Mycoplasma pneumoniae; Characteristic of the microbe, mechanisms of pathogenesis and immunological response of man to M pneumoniae infection (in Polish). Post Mikrobiol 1998, 37, 261–271.
  10. Bencina D, Dove P, Muelle−Premru M, Avsic−Zupanc T, Socan M, Beovic B, Arnez M, Narat M: Intrathecal synthesis of specific antibodies in patients with invasion of the central nervous system by Mycoplasma pneumoniae. Eur J Clin Microbiol Infect Dis 2000, 19, 521–530.
  11. Cassell GH, Waites KB, Pate MS, Canupp KC, Duffy LB: Comparative susceptibility of Mycoplasma pneumoniae to erythromycin, ciprofloxacin and lomefloxacin. Diagn Microbiol Infect Dis 1989, 12, 433–435.
  12. Waites KB, Duffy LB, Schmid T, Crabb D, Pate MS, Cassell GH: In vitro susceptibilities of Mycoplasma pneumoniae, Mycoplasma hominis and Ureaplasma urealyticum to sparfloxacin and PD 127391. Antimicrob Agents Chemother 1991, 6, 1181–1187.
  13. Rennie KA, Prasad ES, Wenman WN: In vitro activity of Dirithromycin, a new macrolide antibiotic, against Mycoplasma species. Diagn Microbiol Infect Dis 1994, 20, 57–61.
  14. Lind K, Madsen H, Wiik A: Autoantibodies to the mitotic spindle apparatus in Mycoplasma pneumoniae disease. Infect Immun 1998, 56, 714–715.
  15. Jacobs E: Serological diagnosis of Mycoplasma pneumoniae infections: a critical review of current procedures. Clin Infect Dis 1993, 17 (Suppl.1), abstr S79–S82.
  16. Tuuminen T, Suni J, Kleemola M, Jacobs E: Improved sensitivity and specificity of enzyme immunoassays with P1−adhesin enriched antigen to detect acute Mycoplasma pneumoniae infection. J Microbiol Methods 2001, 1, 44, 27–37.
  17. Sillis M: The limitations of IgM assays in the serological diagnosis of Mycoplasma pneumoniae infections. J Med Microbiol 1990, 33, 253–258.
  18. Thacker WL, Talkington DF: Analysis of complement fixation and commercial enzyme immunoassays for detection of antibodies to Mycoplasma pneumoniae in human serum. Clin Diagn Labor Immunol 2000, 7, 778–787.
  19. Lüneberg E, Jensen JS, Frosch M: Detection of Mycoplasma pneumoniae by polymerase chain reaction and nonradioactive hybridization in microtiter plates. J Clin Microbiol 1993, 31, 1088–1094.
  20. Blackmore TK, Reznikov M, Gordon DL: Clinical utility of the polymerase chain reaction to diagnose Mycoplasma pneumoniae infection. Pathology 1995, 27, 177–181.
  21. Grattard F, Bourlet T, Galambrun C, Berger C, Stephan JL, Lauras B, Pozzetto B: Interest of gene amplification by PCR for the diagnosis of Mycoplasma pneumoniae infections in the child. Pathol Biol 1998, 46, 464–469.
  22. Rastawicki W, Jagielski M, Gierczyński R: Application of PCR for detection of the selected Mycoplasma pneumoniae genes (in Polish). Med Dośw Mikrobiol 2000, 52, 67–74.
  23. Matas L, Dominquez J, De Ory F, Garcia N, Cardona PJ, Hernandez A, Rodrigo C, Ausina V: Evaluation of Meridian ImmunoCard Mycoplasma test for the detection of Mycoplasma pneumoniae−specific IgM in pediatric patients. Scand J Infect Dis 1998, 30, 289–293.
  24. Mączyńska B, Chiciak J, Stankiewicz M, Przondo−Mordarska A: Detectability of Mycoplasma pneumoniae antibodies using different serological methods in children with atypical pneumonia (in Polish). Adv Clin Exp Med 2002, 11, 451–456.
  25. Cimolai N, Cheong ACH: An assessment of new diagnosis indirect enzyme immunoassay for the detection of anti−Mycoplasma pneumoniae IgM. Am J Clin Pathol 1996, 105, 205–209.
  26. Rastawicki W, Jagielski M: Mycoplasma pneumoniae. II. Clinical features, epidemiology and diagnosis of M. pneumoniae infections (in Polish). Post Mikrobiol 1998, 37, 273–288.
  27. Kai M, Kamiya S, Yale H, Tkatura I, Shiozawa K, Ozawa A: Rapid detection of Mycoplasma pneumoniae in clinical samples by the polymerase chain reaction. J Med Microbiol 1993, 38, 166–170.
  28. Haima P, Oerdijk M, Loens K, Ieven M, Van Roosmalen I, Sillekens P: Molecular diagnosis of Mycoplasma pneumoniae infections by using NASBA with “Real time” detection. Clin Microbol Infect 2001, 8 (Suppl. 1), abstr. P731: 148.