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Chapter 10.6 : Isolation of spp. in Cell Culture

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Isolation of spp. in Cell Culture, Page 1 of 2

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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 . Nucleic acid amplification tests (NAAT) for the detection of genital infections with have largely replaced cell culture. This is for a variety of reasons, including their sensitivity, ease of specimen collection with urine and vaginal swabs being acceptable, and convenient specimen transport. However, cell culture may still be preferred or recommended for certain situations such as laboratory testing in cases of sexual abuse, medicolegal situations, pneumonia, and ophthalmia neonatorum ( ).

Citation: Leber A. 2016. Isolation of spp. in Cell Culture, p 10.6.1-10.6.11. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.6
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Figures

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Figure 10.6–1

Developmental cycle of spp.

Citation: Leber A. 2016. Isolation of spp. in Cell Culture, p 10.6.1-10.6.11. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.6
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Image of Figure 10.6–2
Figure 10.6–2

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

Citation: Leber A. 2016. Isolation of spp. in Cell Culture, p 10.6.1-10.6.11. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.6
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References

/content/book/10.1128/9781555818814.chap10.6
1. CDC. 2010. Sexually Transmitted Diseases Treatment Guidelines, 2010. Morb Mortal Wkly Rep 59:1109.
2. Aarnaes SL, Peterson EM, de la Maza LM. 1984. The effect of media and temperature on the storage of Chlamydia trachomatis. Am J Clin Pathol 81:237239.
3. Lees MI, Newnan DM, Plackette H, Tray-nor PW, Forsyth JR, Garland SM. 1990. A comparison of cytobrush and cotton swab sampling for the detection of Chlamydia trachomatis by cell culture. Genitourin Med 66:267269.
4. Gaydos C, Essig A. 2011. Chlamydiaceae and Chlamydophila, p 9861000. In Versalovic J, Carroll KC, Funke G, Jorgensen JH, Landry ML, Warnock DW (ed), Manual of Clinical Microbiology, 10th ed. ASM Press, Washington, DC.
5. Cles LD, Stamm WE. 1990. Use of HL cells for improved isolation and passage of Chlamydia pneumoniae. J Clin Microbiol 28:938940.
6. Roblin PM, Dumornay W, Hammerschlag MR. 1992. Use of HEp-2 cells for improved isolation and passage of Chlamydia pneumoniae. J Clin Microbiol 30:19681971.
7. Barnes RC. 1989. Laboratory diagnosis of human chlamydial infections. Clin Microbiol Rev 2:119136.
8. Mahony JB, Chernesky MA. 1985. Effect of swab type and storage temperature on the isolation of Chlamydia trachomatis from clinical specimens. J Clin Microbiol 22:865867.
9. Pruckler JM, Masse N, Stevens VA, Gang L, Yang Y, Zell ER, Dowell SF, Fields BS. 1999. Optimizing the culture of Chlamydia pneumoniae by using multiple centrifugations. J Clin Microbiol 37:33993401.
10. Smith TF. 1982. Role of the diagnostic virology laboratory in clinical microbiology: tests for Chlamydia trachomatis and enteric toxins in cell culture, p 82119. In de la Maza LM, Peterson EM (ed), Medical Virology. Elsevier Biomedical Press, New York, NY.
11. Bird BR, Forrester FT. 1981. Laboratory Diagnosis of Chlamydial Infections, p 5562. CDC, Atlanta, GA.
12. CDC. 1990. False-positive results with the use of chlamydia tests in the evaluation of suspected sexual abuse. Morb Mortal Wkly Rep 39:932935.
13. Kuo CC. Wang SP, Wentworth BB, Grayston JT. 1972. Primary isolation of TRIC organisms in HeLa 229 cells treated with DEAE-dextran. J Infect Dis 125:665668.
1. Cles LD, Stamm WE. 1990. Use of HL cells for improved isolation and passage of Chlamydia pneumoniae. J Clin Microbiol 28:938940.
2. Ripa KT, Mardh PA. 1977. New simplified culture technique for Chlamydia trachomatis, p 323327. In Holmes KK, Hobson D (ed), Non-Gonococcal Urethritis and Related Infections. American Society for Microbiology, Washington, DC.
3. Ripa KT, Mardh PA. 1977. Cultivation of Chlamydia trachomatis in cycloheximide-treated McCoy cells. J Clin Microbiol 6:328331.
4. Kuo CC, Wang SP, Wentworth BB, Grayston JT. 1972. Primary isolation of TRIC organisms in HeLa 229 cells treated with DEAE-dextran. J Infect Dis 125:665668.
1. de la Maza LM, Peterson EM. 1981. Scanning electron microscopy of McCoy cells infected with Chlamydia trachomatis. Exp Mol Pathol 36:217226.
2. Schachter J, Dawson C. 1978. Human Chlamydial Infections. Publishing Sciences Group, Littleton, MA.
3. Munday PE, Johnson AP, Thomas BJ, Taylor-Robinson D. 1980. A comparison of immunofluorescence and Giemsa for staining Chlamydia trachomatis inclusions in cycloheximide-treated McCoy cells. J Clin Pathol 33:177179.
4. Schachter J. 1985. Immunodiagnosis of sexually transmitted disease. Yale J Biol Med 58:443452.

Tables

Generic image for table
Table 10.6–1

Human chlamydial infections

Citation: Leber A. 2016. Isolation of spp. in Cell Culture, p 10.6.1-10.6.11. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.6
Generic image for table
Table 10.6–2

Collection of specimens

Citation: Leber A. 2016. Isolation of spp. in Cell Culture, p 10.6.1-10.6.11. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.6
Generic image for table
Table 10.5–A1

Problems associated with interpretation of the neutralization test

Citation: Leber A. 2016. Isolation of spp. in Cell Culture, p 10.6.1-10.6.11. In Clinical Microbiology Procedures Handbook, Fourth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555818814.ch10.6

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