Mycoplasma: Immunologic and Molecular Diagnostic Methods

Ken B. Waites; Mary B. Brown; Jerry W. Simecka

doi:10.1128/9781555818722.ch49

Chapter 49 : Mycoplasma: Immunologic and Molecular Diagnostic Methods

Authors: Ken B. Waites¹, Mary B. Brown², Jerry W. Simecka³

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Affiliations: 1: Department of Pathology, University of Alabama at Birmingham, WP 230, 619 S. 19th St., Birmingham, AL 35249; 2: Department of Infectious Diseases and Pathology, College of Veterinary Medicine, University of Florida, P.O. Box 110880, 2015 S.W. 16th Ave., Gainesville, FL 32611; 3: Department of Cell Biology and Immunology, University of North Texas Health Science Center, RES 402A 3500 Camp Bowie Blvd., Fort Worth, TX 76107

Content Type: Reference

Publication Year: 2016

Category: Immunology

Book DOI: 10.1128/9781555818722

Chapter DOI: 10.1128/9781555818722.ch49

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Abstract:

Mycoplasmas and ureaplasmas are members of a unique group of organisms (class Mollicutes) that are characterized by their small genomes, lack of cell walls, sterols in cell membranes, and complex nutritional requirements. The role of Mycoplasma and Ureaplasma species in human diseases was largely underappreciated until recent years. As a result, most diagnostic laboratories ignored them. Because of their complex nutritional requirements as well as their adaptation to the host during infection, these fastidious organisms can be difficult and time-consuming to culture from patient samples. However, there are improved methods for detection, including PCR detection assays, serologic assays, and commercially available growth media, but these are still limited compared with the products for other organisms.

Citation: Waites K, Brown M, Simecka J. 2016. Mycoplasma: Immunologic and Molecular Diagnostic Methods, p 444-452. In Detrick B, Schmitz J, Hamilton R (ed), Manual of Molecular and Clinical Laboratory Immunology, Eighth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818722.ch49

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References

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1. Waites KB, Xiao L, Paralanov V, Viscardi RM, Glass JI, . 2013. Mycoplasma and Ureaplasma, p 229– 281. In de Filippis I, McKee ML (ed), Molecular Typing in Bacterial Infections. Springer Science, New York, NY.

2. Waites KB, Talkington DF . 2004. Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev 17 : 697– 728.[CrossRef].[PubMed]

3. Narita M . 2005. [Evaluation of ELISA kits for detection of Mycoplasma pneumoniae-specific IgG, IgA, IgM antibodies on the diagnosis of Mycoplasma pneumoniae infection in children]. Kansenshogaku Zasshi 79 : 457– 463.[PubMed].[CrossRef]

4. Loens K, Goossens H, Ieven M . 2010. Acute respiratory infection due to Mycoplasma pneumoniae: current status of diagnostic methods. Eur J Clin Microbiol Infect Dis 29 : 1055– 1069.[CrossRef].[PubMed]

5. Beersma MF, Dirven K, van Dam AP, Templeton KE, Claas EC, Goossens H . 2005. Evaluation of 12 commercial tests and the complement fixation test for Mycoplasma pneumoniae-specific immunoglobulin G (IgG) and IgM antibodies, with PCR used as the “gold standard.” J Clin Microbiol 43 : 2277– 2285.[CrossRef].[PubMed]

6. Busson L, Van den Wijngaert S, Dahma H, Decolvenaer M, Di Cesare L, Martin A, Vasseur L, Vandenberg O . 2013. Evaluation of 10 serological assays for diagnosing Mycoplasma pneumoniae infection. Diagn Microbiol Infect Dis 76 : 133– 137.[CrossRef].[PubMed]

7. Talkington DF, Shott S, Fallon MT, Schwartz SB, Thacker WL . 2004. Analysis of eight commercial enzyme immunoassay tests for detection of antibodies to Mycoplasma pneumoniae in human serum. Clin Diagn Lab Immunol 11 : 862– 867.[CrossRef].[PubMed]

8. Petitjean J, Vabret A, Gouarin S, Freymuth F . 2002. [Evaluation of four commercial immunoglobulin G (IgG)- and IgM-specific enzyme immunoassays for diagnosis of Mycoplasma pneumoniae infections]. Pathol Biol (Paris) 50 : 530– 537.[PubMed].[CrossRef]

9. Ozaki T, Nishimura N, Ahn J, Watanabe N, Muto T, Saito A, Koyama N, Nakane K, Funahashi K . 2007. Utility of a rapid diagnosis kit for Mycoplasma pneumoniae pneumonia in children, and the antimicrobial susceptibility of the isolates. J Infect Chemother 13 : 204– 207.[CrossRef].[PubMed]

10. Csango PA, Pedersen JE, Hess RD . 2004. Comparison of four Mycoplasma pneumoniae IgM-, IgG- and IgA-specific enzyme immunoassays in blood donors and patients. Clin Microbiol Infect 10 : 1094– 1098.[CrossRef].[PubMed]

11. Pierce VM, Elkan M, Leet M, McGowan KL, Hodinka RL . 2012. Comparison of the Idaho Technology FilmArray system to real-time PCR for detection of respiratory pathogens in children. J Clin Microbiol 50 : 364– 371.[CrossRef].[PubMed]

12. Loens K, van Loon AM, Coenjaerts F, van Aarle Y, Goossens H, Wallace P, Claas E J, Ieven M . 2012. Performance of different mono- and multiplex nucleic acid amplification tests on a multipathogen external quality assessment panel. J Clin Microbiol 50 : 977– 987.[CrossRef].[PubMed]

13. Waites KB, Xiao L, Paralanov V, Viscardi RM, Glass JI . 2012. Molecular methods for the detection of Mycoplasma and Ureaplasma infections in humans: a paper from the 2011 William Beaumont Hospital Symposium on molecular pathology. J Mol Diagn 14 : 437– 450.[CrossRef].[PubMed]

14. Touati A, Benard A, Hassen AB, Bebear CM, Pereyre S . 2009. Evaluation of five commercial real-time PCR assays for detection of Mycoplasma pneumoniae in respiratory tract specimens. J Clin Microbiol 47 : 2269– 2271.[CrossRef].[PubMed]

15. Dumke R, Jacobs E . 2009. Comparison of commercial and in-house real-time PCR assays used for detection of Mycoplasma pneumoniae. J Clin Microbiol 47 : 441– 444.[CrossRef].[PubMed]

16. Ursi D, Ieven M, Noordhoek GT, Ritzler M, Zandleven H, Altwegg M . 2003. An interlaboratory comparison for the detection of Mycoplasma pneumoniae in respiratory samples by the polymerase chain reaction. J Microbiol Methods 53 : 289– 294.[PubMed].[CrossRef]

17. Loens K, Beck T, Ursi D, Pattyn S, Goossens H, Ieven M . 2006. Two quality control exercises involving nucleic acid amplification methods for detection of Mycoplasma pneumoniae and Chlamydophila pneumoniae and carried out 2 years apart (in 2002 and 2004). J Clin Microbiol 44 : 899– 908.[CrossRef].[PubMed]

18. Furr PM, Taylor-Robinson D . 1984. Microimmunofluorescence technique for detection of antibody to Mycoplasma genitalium. J Clin Pathol 37 : 1072– 1074.[PubMed].[CrossRef]

19. Baseman JB, Cagle M, Korte JE, Herrera C, Rasmussen WG, Baseman JG, Shain R, Piper JM . 2004. Diagnostic assessment of Mycoplasma genitalium in culture-positive women. J Clin Microbiol 42 : 203– 211.[PubMed].[CrossRef]

20. Jurstrand M, Jensen JS, Fredlund H, Falk L, Molling P . 2005. Detection of Mycoplasma genitalium in urogenital specimens by real-time PCR and by conventional PCR assay. J Med Microbiol 54 : 23– 29.[CrossRef].[PubMed]

21. Deguchi T, Yoshida T, Yokoi S, Ito M, Tamaki M, Ishiko H, Maeda S . 2002. Longitudinal quantitative detection by real-time PCR of Mycoplasma genitalium in first-pass urine of men with recurrent nongonococcal urethritis. J Clin Microbiol 40 : 3854– 3856.[PubMed].[CrossRef]

22. Jensen JS, Bjornelius E, Dohn B, Lidbrink P . 2004. Use of TaqMan 5′ nuclease real-time PCR for quantitative detection of Mycoplasma genitalium DNA in males with and without urethritis who were attendees at a sexually transmitted disease clinic. J Clin Microbiol 42 : 683– 692.[PubMed].[CrossRef]

23. Svenstrup HF, Jensen JS, Bjornelius E, Lidbrink P, Birkelund S, Christiansen G . 2005. Development of a quantitative real-time PCR assay for detection of Mycoplasma genitalium. J Clin Microbiol 43 : 3121– 3128.[CrossRef].[PubMed]

24. Dupin N, Bijaoui G, Schwarzinger M, Ernault P, Gerhardt P, Jdid R, Hilab S, Pantoja C, Buffet M, Escande JP, Costa J M . 2003. Detection and quantification of Mycoplasma genitalium in male patients with urethritis. Clin Infect Dis 37 : 602– 605.[CrossRef].[PubMed]

25. Grau O, Kovacic R, Griffais R, Launay V, Montagnier L . 1994. Development of PCR-based assays for the detection of two human mollicute species, Mycoplasma penetrans and M. hominis. Mol Cell Probes 8 : 139– 147.[CrossRef].[PubMed]

26. Yoshida T, Maeda S, Deguchi T, Miyazawa T, Ishiko H . 2003. Rapid detection of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum organisms in genitourinary samples by PCR-microtiter plate hybridization assay. J Clin Microbiol 41 : 1850– 1855.[PubMed].[CrossRef]

27. Ferandon C, Peuchant O, Janis C, Benard A, Renaudin H, Pereyre S, Bebear C . 2011. Development of a real-time PCR targeting the yidC gene for the detection of Mycoplasma hominis and comparison with quantitative culture. Clin Microbiol Infect 17 : 155– 159.[CrossRef].[PubMed]

28. Baczynska A, Svenstrup HF, Fedder J, Birkelund S, Christiansen G . 2004. Development of real-time PCR for detection of Mycoplasma hominis. BMC Microbiol 4 : 35.[CrossRef].[PubMed]

29. Menard JP, Fenollar F, Henry M, Bretelle F, Raoult D . 2008. Molecular quantification of Gardnerella vaginalis and Atopobium vaginae loads to predict bacterial vaginosis. Clin Infect Dis 47 : 33– 43.[CrossRef].[PubMed]

30. Cunningham SA, Mandrekar JN, Rosenblatt JE, Patel R . 2013. Rapid PCR detection of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum. Int J Bacteriol 2013 : 7.

31. Maeda S, Deguchi T, Ishiko H, Matsumoto T, Naito S, Kumon H, Tsukamoto T, Onodera S, Kamidono S . 2004. Detection of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum (biovar 1) and Ureaplasma urealyticum (biovar 2) in patients with non-gonococcal urethritis using polymerase chain reaction-microtiter plate hybridization. Int J Urol 11 : 750– 754.[CrossRef].[PubMed]

32. Xiao L, Glass JI, Paralanov V, Yooseph S, Cassell GH, Duffy LB, Waites KB . 2010. Detection and characterization of human Ureaplasma species and serovars by real-time PCR. J Clin Microbiol 48 : 2715– 2723.[CrossRef].[PubMed]

33. Povlsen K, Jensen JS, Lind I . 1998. Detection of Ureaplasma urealyticum by PCR and biovar determination by liquid hybridization. J Clin Microbiol 36 : 3211– 3216.[PubMed]

34. McKechnie ML, Hillman R, Couldwell D, Kong F, Freedman E, Wang H, Gilbert GL . 2009. Simultaneous identification of 14 genital microorganisms in urine by use of a multiplex PCR-based reverse line blot assay. J Clin Microbiol 47 : 1871– 1877.[CrossRef].[PubMed]

35. Wang Y, Kong F, Gilbert GL, Brown M, Gao W, Yu S, Yang Y . 2008. Use of a multiplex PCR-based reverse line blot (mPCR/RLB) hybridisation assay for the rapid identification of bacterial pathogens. Clin Microbiol Infect 14 : 155– 160.[CrossRef].[PubMed]

36. Wang H, Kong F, Jelfs P, James G, Gilbert GL . 2004. Simultaneous detection and identification of common cell culture contaminant and pathogenic mollicutes strains by reverse line blot hybridization. Appl Environ Microbiol 70 : 1483– 1486.[PubMed].[CrossRef]

37. Benstein BD, Crouse DT, Shanklin DR, Ourth DD . 2003. Ureaplasma in lung. 1. Localization by in situ hybridization in a mouse model. Exp Mol Pathol 75 : 165– 170.[PubMed].[CrossRef]

38. Pitcher D, Chalker VJ, Sheppard C, George RC, Harrison TG . 2006. Real-time detection of Mycoplasma pneumoniae in respiratory samples with an internal processing control. J Med Microbiol 55 : 149– 155.[CrossRef].[PubMed]

39. Saito R, Misawa Y, Moriya K, Koike K, Ubukata K, Okamura N . 2005. Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of Mycoplasma pneumoniae. J Med Microbiol 54 : 1037– 1041.[CrossRef].[PubMed]

40. Di Marco E, Cangemi G, Filippetti M, Melioli G, Biassoni R . 2007. Development and clinical validation of a real-time PCR using a uni-molecular Scorpion-based probe for the detection of Mycoplasma pneumoniae in clinical isolates. New Microbiol 30 : 415– 421.[PubMed]

41. Ginevra C, Barranger C, Ros A, Mory O, Stephan JL, Freymuth F, Joannes M, Pozzetto B, Grattard F . 2005. Development and evaluation of Chlamylege, a new commercial test allowing simultaneous detection and identification of Legionella, Chlamydophila pneumoniae, and Mycoplasma pneumoniae in clinical respiratory specimens by multiplex PCR. J Clin Microbiol 43 : 3247– 3254.[CrossRef].[PubMed]

42. Dumke R, Schurwanz N, Lenz M, Schuppler M, Luck C, Jacobs E . 2007. Sensitive detection of Mycoplasma pneumoniae in human respiratory tract samples by optimized real-time PCR approach. J Clin Microbiol 45 : 2726– 2730.[CrossRef].[PubMed]

43. Winchell JM, Thurman KA, Mitchell SL, Thacker W, Fields BS . 2008. Evaluation of three real-time PCR assays for the detection of Mycoplasma pneumoniae in an outbreak investigation. J Clin Microbiol. 46 : 3116– 3118.[CrossRef].[PubMed]

44. Stormer M, Vollmer T, Henrich B, Kleesiek K, Dreier J . 2009. Broad-range real-time PCR assay for the rapid identification of cell-line contaminants and clinically important mollicute species. Int J Med Microbiol 299 : 291– 300.[CrossRef].[PubMed]

45. Khanna M, Fan J, Pehler-Harrington K, Waters C, Douglass P, Stallock J, Kehl S, Henrickson KJ . 2005. The pneumoplex assays, a multiplex PCR-enzyme hybridization assay that allows simultaneous detection of five organisms, Mycoplasma pneumoniae, Chlamydia ( Chlamydophila) pneumoniae, Legionella pneumophila, Legionella micdadei, and Bordetella pertussis, and its real-time counterpart. J Clin Microbiol 43 : 565– 571.[CrossRef].[PubMed]

46. Morozumi M, Nakayama E, Iwata S, Aoki Y, Hasegawa K, Kobayashi R, Chiba N, Tajima T, Ubukata K . 2006. Simultaneous detection of pathogens in clinical samples from patients with community-acquired pneumonia by real-time PCR with pathogen-specific molecular beacon probes. J Clin Microbiol 44 : 1440– 1446.[CrossRef].[PubMed]

47. Raggam RB, Leitner E, Berg J, Muhlbauer G, Marth E, Kessler HH . 2005. Single-run, parallel detection of DNA from three pneumonia-producing bacteria by real-time polymerase chain reaction. J Mol Diagn 7 : 133– 138.[CrossRef].[PubMed]

48. Loens K, Ieven M, Ursi D, Beck T, Overdijk M, Sillekens P, Goossens H . 2003. Detection of Mycoplasma pneumoniae by real-time nucleic acid sequence-based amplification. J Clin Microbiol 41 : 4448– 4450.[PubMed].[CrossRef]

49. Lin B, Blaney KM, Malanoski AP, Ligler AG, Schnur JM, Metzgar D, Russell KL, Stenger DA . 2007. Using a resequencing microarray as a multiple respiratory pathogen detection assay. J Clin Microbiol 45 : 443– 452.[CrossRef].[PubMed]

50. Yoshida T, Deguchi T, Meda S, Kubota Y, Tamaki M, Yokoi S, Yasuda M, Ishiko H . 2007. Quantitative detection of Ureaplasma parvum (biovar 1) and Ureaplasma urealyticum (biovar 2) in urine specimens from men with and without urethritis by real-time polymerase chain reaction. Sex Transm Dis 34 : 416– 419.[CrossRef].[PubMed]

51. Mallard K, Schopfer K, Bodmer T . 2005. Development of real-time PCR for the differential detection and quantification of Ureaplasma urealyticum and Ureaplasma parvum. J Microbiol Methods 60 : 13– 19.[CrossRef].[PubMed]

52. Cao X, Wang Y, Hu X, Qing H, Wang H . 2007. Real-time TaqMan polymerase chain reaction assays for quantitative detection and differentiation of Ureaplasma urealyticum and Ureaplasma parvum. Diagn Microbiol Infect Dis 57 : 373– 378.[CrossRef].[PubMed]

53. Vancutsem E, Soetens O, Breugelmans M, Foulon W, Naessens A . 2011. Modified real-time PCR for detecting, differentiating, and quantifying Ureaplasma urealyticum and Ureaplasma parvum. J Mol Diagn 13 : 206– 212.[CrossRef].[PubMed]

54. Yoshida T, Deguchi T, Ito M, Maeda S, Tamaki M, Ishiko H . 2002. Quantitative detection of Mycoplasma genitalium from first-pass urine of men with urethritis and asymptomatic men by real-time PCR. J Clin Microbiol 40 : 1451– 1455.[PubMed].[CrossRef]

55. Yoshida T, Maeda S, Deguchi T, Ishiko H . 2002. Phylogeny-based rapid identification of mycoplasmas and ureaplasmas from urethritis patients. J Clin Microbiol 40 : 105– 110.[PubMed].[CrossRef]

56. Pascual A, Jaton K, Ninet B, Bille , Greub G . 2010. New diagnostic real-time PCR for specific detection of Mycoplasma hominis DNA. Int J Bacteriol 2010 : 4.

Tables

Mycoplasma: Immunologic and Molecular Diagnostic Methods

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/content/10.1128/9781555818722.ch49.ch49tab1

TABLE 1

Major commercial serologic test formats for M. pneumoniae in the United States

TABLE 1

Major commercial serologic test formats for M. pneumoniae in the United States

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TABLE 2

Examples of M. pneumoniae nucleic acid amplification assays

TABLE 2

Examples of M. pneumoniae nucleic acid amplification assays

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TABLE 3

Examples of real-time PCR-based assays and gene targets for detection of urogenital mycoplasmas

TABLE 3

Examples of real-time PCR-based assays and gene targets for detection of urogenital mycoplasmas