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Chapter 3.15 : Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR

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Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, Page 1 of 2

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Abstract:

is a common cause of upper and lower respiratory infections in persons of all ages. Tracheobronchitis is the most common clinical syndrome, but pneumonia commonly occurs and extrapulmonary infections have been described. More detailed information concerning human diseases and clinical aspects of infections caused by is available in reference texts and reviews ( ). Specimens obtained from the upper and lower respiratory tracts are appropriate to culture for the presence of . Since the organism has also been known to cause invasive disease in other body sites, it may be appropriate in some circumstances to culture sterile body fluids such as CSF, synovial fluid, or other types of clinical specimens, depending on the type of infection suspected. Growth and presumptive identification of can be accomplished by demonstration of glucose utilization in SP4 broth and development of spherical 10- to 100-μm colonies on SP4 agar after 4 to 20 days or more of incubation. Definitive organism identification has been accomplished using a variety of tests such as hemadsorption, tetrazolium reduction, or growth inhibition in the presence of specific antiserum. More recently, the PCR assay has been used to distinguish isolated in culture from several species of commensal mycoplasmas that often inhabit the human respiratory tract ( ). Since culture may not detect in some specimens and requires several days to weeks to complete, alternative methods such as molecular assays and serologic testing should be considered for optimum detection, even if culture is attempted.

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
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Figures

Image of Figure 3.15.1–1
Figure 3.15.1–1

Algorithm for isolation of from clinical specimens. Rate of growth, glucose metabolism, colonial morphology, and simple confirmatory tests can usually differentiate from other mycoplasmas and acholeplasmas that may be present in the respiratory tract. Noting the presence of a mycoplasma other than will allow clinicians to determine whether further characterization is needed. *, Acidic or alkaline shift with turbidity indicates contamination and overgrowth with other bacteria or yeast.

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
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Image of Figure 3.15.1–2
Figure 3.15.1–2

Algorithm for isolation of and spp. from clinical specimens. is the most common mycoplasmal species isolated from the genitourinary tract or from extragenital sites. Preliminary differentiation of from other potentially pathogenic mycoplasmas can usually be made based on a combination of colony morphology, growth rate, and differential utilization of glucose and/or arginine. Rare isolations of other human mycoplasmas, or species not normally associated with humans, can cause confusion, thereby necessitating speciation procedures outside the purview of most clinical laboratories. *, Acidic or alkaline shift with turbidity indicates contamination and overgrowth with other bacteria or yeast.

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
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Image of Figure 3.15.1–3
Figure 3.15.1–3

Spherical colonies of up to 100 μm in diameter growing on SP4 agar. Magnification, ×126.

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
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Image of Figure 3.15.1–4
Figure 3.15.1–4

Small colonies typical of spp. and larger fried-egg colonies of Diene’s stain; magnification, ×100. Reprinted from M. G. Gabridge. 1981. , part 2, no. 6702 (teaching set). American Society for Microbiology, Washington, DC.

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
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References

/content/book/10.1128/9781555818814.chap3.15
1. Waites KB, Balish MF, Atkinson TP. 2008. New insights into the pathogenesis and detection of Mycoplasma pneumoniae infections. Future Microbiol 3:635648.
2. Waites KB, Taylor-Robinson D. 2011. Mycoplasma and Ureaplasma, p 970985. In Versalovic J, Carroll KC, Jorgensen JG, Funke G, Landry ML, Warnock DW (ed), Manual of Clinical Microbiology, 10th ed. ASM Press, Washington, DC.
3. 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 14:437450.
4. Waites KB, Katz B, Schelonka RL. 2005. Mycoplasmas and ureaplasmas as neonatal pathogens. Clin Microbiol Rev 18:757789.
5. Taylor-Robinson D, Jensen JS. 2011. Mycoplasma genitalium: from chrysalis to multicolored butterfly. Clin Microbiol Rev 24:498514.
1. CLSI. 2004. Quality Control for Commercially Prepared Microbiological Culture Media. Approved standard M22-A3, 3rd ed. CLSI, Wayne, PA.
1. Dumke R, Schurwanz N, Lenz M, Schuppler C, Luck C, Jacobs E. 2007. Sensitive detection of Mycoplasma pneumoniae in human respiratory tract samples by optimized real-time PCR approach. J Clin Microbiol 45:27262730.
1. 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 43:31213128.
1. Cunningham SE, Mandrekar JN, Rosenblatt JE, Patel R. 2013. Rapid PCR Detection of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum. Int J Bacteriol 2013:168742. doi:10.1155/2013/168742.
1. Waites KB, Katz B, Schelonka RL. 2005. Mycoplasmas and ureaplasmas as neonatal pathogens. Clin Microbiol Rev 18:757789.
2. 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 48:27152723.
1. Waites KB, Bebear CM, Robertson JA, Talkington DF, Kenny GE. 2001. Cumitech 34, Laboratory diagnosis of mycoplasmal infections. Coordinating ed, Nolte F. ASM Press, Washington, DC.
1. CLSI. 2011. Methods for Antimicrobial Susceptibility Testing of Human Mycoplasmas. Approved guideline. CLSI document M43-A. CLSI, Wayne, PA.
2. CLSI. 2015. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically, 10th ed. Approved standard M7-A10. CLSI, Wayne, PA.

Tables

Generic image for table
Mpn repMp1 Program

Mpn repMp1 Program

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
Generic image for table
MGen-GAP Program

MGen-GAP Program

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
Generic image for table
MHtuf Program

MHtuf Program

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
Generic image for table
UP and UU Multiplex Program

UP and UU Multiplex Program

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
Generic image for table
Untitled

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15
Generic image for table
MIC QC ranges for Mycoplasma hominis, Mycoplasma pneumoniae, and Ureaplasma urealyticum (broth microdilution method from CLSI M43-A [1])

MIC QC ranges for and (broth microdilution method from CLSI M43-A [ ])

Citation: Waites K, Duffy L, Xiao L. 2016. Detection of Human Mycoplasmas and Ureaplasmas from Clinical Specimens by Culture and PCR, p 3.15.1.1-3.15.8.5. In Leber A (ed), Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch3.15

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