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Chapter 10.6 : Isolation of Chlamydia spp. in Cell Culture

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Abstract:

Chlamydiae are obligate intracellular bacteria that contain RNA and DNA, have a cell wall resembling those of gram-negative bacteria, and multiply by binary fission in a manner distinct from those of other bacteria. The 300- to 400-nm spherical elementary body (EB) is the infectious form of the organism. Following cellular infection, the EB reorganizes into a larger, metabolically active reticulate body (RB), which divides repeatedly by binary fission for 24 to 48 h and eventually develops into the characteristic intracytoplasmic inclusion. Human infections associated with the genus are summarized in Table 10.6-1 . Despite the introduction of numerous nonculture assays, including amplified assays, culture remains an important assay for the detection of infections and, because of its specificity, is recommended for laboratory testing in cases of sexual abuse and medicolegal situations.

Citation: Garcia L. 2010. Isolation of Chlamydia spp. in Cell Culture, p 92-103. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.6
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Figures

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Figure 10.6-1

Developmental cycle of spp.

Citation: Garcia L. 2010. Isolation of Chlamydia spp. in Cell Culture, p 92-103. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.6
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Image of Figure 10.6-2
Figure 10.6-2

Typical chlamydial inclusions in a culture stained at 48 h with a onoclonal antibody to

Citation: Garcia L. 2010. Isolation of Chlamydia spp. in Cell Culture, p 92-103. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.6
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References

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1. Aarnaes, S. L.,, E. M. Peterson,, and L. M. de la Maza. 1984. The effect of media and temperature on the storage of Chlamydia trachomatis. Am. J. Clin. Pathol. 81:237239.
2. Barnes, R. C. 1989. Laboratory diagnosis of human chlamydial infections. Clin. Microbiol. Rev. 2:119136.
3. Bird, B. R.,, and F. T. Forrester. 1981. Laboratory Diagnosis of Chlamydial Infections, p. 5562. Centers for Disease Control, Atlanta, Ga.
4.Centers for Disease Control. 1990. Falsepositive results with the use of chlamydia tests in the evaluation of suspected sexual abuse. Morb. Mortal. Wkly. Rep. 39:932935.
5. Cles, L. D.,, and W. E. Stamm. 1990. Use of HL cells for improved isolation and passage of Chlamydia pneumoniae. J. Clin. Microbiol. 28:938940.
6. Kuo, C.-C.,, S. P. Wang,, B. B. Wentworth,, and J. T. Grayston. 1972. Primary isolation of TRIC organisms in HeLa 229 cells treated with DEAE-dextran. J. Infect. Dis. 125:665668.
7. Lees, M. I.,, D. M. Newnan,, H. Plackette,, P. W. Traynor,, J. R. Forsyth,, and S. M. Garland. 1990. A comparison of cytobrush and cotton swab sampling for the detection of Chlamydia trachomatis by cell culture. Genitourin. Med. 66:267269.
8. Mahony, J. B.,, and M. A. Chernesky. 1985. Effect of swab type and storage temperature on the isolation of Chlamydia trachomatis from clinical specimens. J. Clin. Microbiol. 22:865867.
9. Pruckler, J. M.,, N. Masse,, V. A. Stevens,, L. Gang,, Y. Yang,, E. R. Zell,, S. F. Dowell,, and B. S. Fields. 1999. Optimizing the culture of Chlamydia pneumoniae by using multiple centrifugations. J. Clin. Microbiol. 37:33993401.
10. Roblin, P. M.,, W. Dumornay,, and M. R. Hammerschlag. 1992. Use of HEp-2 cells for improved isolation and passage of Chlamydia pneumoniae. J. Clin. Microbiol. 30:19681971.
11. Schachter, J. 1985. Immunodiagnosis of sexually transmitted disease. Yale J. Biol. Med. 58:443452.
12. Smith, T. F., 1982. Role of the diagnostic virology laboratory in clinical microbiology: tests for Chlamydia trachomatis and enteric toxins in cell culture, p. 82119. In L. M. de la Maza, and E. M. Peterson (ed.), Medical Virology. Elsevier Biomedical Press, New York, N.Y.
13. Mahony, J. B.,, B. K. Coombes,, and M. A. Chemesky,. 2003. Chlamydia and Chlamydophila, p.9911004. In P. R. Murray,, E. J. Baron,, J. H. Jorgensen,, M. A. Pfaller,, and R. H. Yolken (ed.), Manual of Clinical Microbiology, 8th ed. ASM Press, Washington, D.C.
1. Cles, L. D.,, and W. E. Stamm. 1990. Use of HL cells for improved isolation and passage of Chlamydia pneumoniae. J. Clin. Microbiol. 28:938940.
2. Kuo, C.-C.,, S. P. Wang,, B. B. Wentworth,, and J. T. Grayston. 1972. Primary isolation of TRIC organisms in HeLa 229 cells treated with DEAE-dextran. J. Infect. Dis. 125:665668.
3. Ripa, K. T.,, and P.-A. Mardh,. 1977. New simplified culture technique for Chlamydia trachomatis, p. 323327. In K. K. Holmes, and D. Hobson (ed.), Non-Gonococcal Urethritis and Related Infections. American Society for Microbiology, Washington, D.C..
4. Ripa, K. T.,, and P.-A. Mardh. 1977. Cultivation of Chlamydia trachomatis in cycloheximide- treated McCoy cells. J. Clin. Microbiol. 6:328331.
1. de la Maza, L. M.,, and E. M. Peterson. 1981. Scanning electron microscopy of McCoy cells infected with Chlamydia trachomatis. Exp. Mol. Pathol. 36:217226.
2. Munday, P. E.,, A. P. Johnson,, B. J. Thomas,, and D. Taylor-Robinson. 1980. A comparison of immunofluorescence and Giemsa for staining Chlamydia trachomatis inclusions in cycloheximide-treated McCoy cells. J. Clin. Pathol. 33:177179.
3. Schachter, J. 1985. Immunodiagnosis of sexually transmitted disease. Yale J. Biol. Med. 58:443452.
4. Schachter, J.,, and C. Dawson. 1978. Human Chlamydial Infections. Publishing Sciences Group, Littleton, Mass.

Tables

Generic image for table
Table 10.6-1

Human chlamydial infections

Citation: Garcia L. 2010. Isolation of Chlamydia spp. in Cell Culture, p 92-103. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.6
Generic image for table
Table 10.6-2

Collection of specimens

Citation: Garcia L. 2010. Isolation of Chlamydia spp. in Cell Culture, p 92-103. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch10.6

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