Comparison of methods for cultivation and isolation of Chlamydia trachomatis

Benes, Solomon; McCormack, William M.

doi:10.1128/jcm.16.5.847-850.1982

Cited by 16 publications

(11 citation statements)

References 18 publications

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“…The combined DEAE-D and cycloheximide treatment resulted in an enhancement of the size of inclusions with little reduction in the number of inclusions. Several investigators have reported on the cycloheximide enhancement of the size of TRIC inclusions in McCoy cells (4,19,20). These investigators have also reported an increase in the number of inclusions detected by iodine staining in the cycloheximidetreated cells.…”

Section: Resultsmentioning

confidence: 99%

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Enhancement of Chlamydia trachomatis infectious progeny by cultivation of HeLa 229 cells treated with DEAE-dextran and cycloheximide

Sabet¹,

Simmons²,

Caldwell³

1984

J Clin Microbiol

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The effects of DEAE-dextran and cycloheximide on the infection of HeLa 229 cells with Chlamydia trachomatis serotype G were studied in terms of the number of cells infected and the yield of infectious progeny per infected cell. Pretreatment of the host cells with DEAE-dextran resulted in an increase in the number of infected cells but had no significant effect on the yield of infectious progeny per infected cell (burst size). In contrast, the addition of cycloheximide to the medium of infected cells had no significant effect on the number of infected cells but greatly enhanced the burst size. The burst size was calculated to be close to 500. The enhanced burst size was also observed in cells treated with DEAE-dextran and cycloheximide. In addition, there was an increase in the number of cells infected and an augmentation of the infectious progeny yield. Under the conditions of combined treatment, the yield of C. trachomatis serotype G cultivated in HeLa 229 cells was found to be approximately threefold higher than the yield of the organisms cultivated in McCoy cells. The results suggest that HeLa 229 cells treated with DEAE-dextran and cycloheximide offer a most suitable system for the high-yield cultivation of C. trachomatis organisms and possibly also for the diagnosis of infection with these organisms. Members of the species Chlamydia trachomatis are gramnegative, spherical bacteria capable of multiplication only inside susceptible mammalian host cells (3). Currently, 15 serotypes of C. trachomatis strains are recognized. Three (Li, L2, and L3) are the lymphogranulo

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Section: Resultsmentioning

confidence: 99%

“…Cycloheximide treatment has been used extensively for the cultivation of TRIC organisms in McCoy cells (4,19,20). However, the effect of this antibiotic on the propagation of TRIC organisms in HeLa 229 has not been investigated.…”

Section: Fig 2 Effects Of Deae-mentioning

confidence: 99%

Enhancement of Chlamydia trachomatis infectious progeny by cultivation of HeLa 229 cells treated with DEAE-dextran and cycloheximide

Sabet¹,

Simmons²,

Caldwell³

1984

J Clin Microbiol

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“…The diagnosis of a C. trachomatis infection, e.g., urethritis or cervicitis, is primarily dependent upon isolation by cell culture procedures. McCoy cells and HeLa 229 cells are commonly used, and various cell treatments have been described to enhance the sensitivity of the assay (1)(2)(3)(4)(5)(6)(7)(8)(9)11). Most authors used one cell type and one or more laboratory strains.…”

mentioning

confidence: 99%

“…However, these strains have had different histories of laboratory passage. The investigators studying the impact of modifications on the isolation rate in clinical specimens (1,2,5,6,9,11) always used one type of cell culture.…”

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Comparison of HeLa 229 and McCoy cell cultures for detection of Chlamydia trachomatis in clinical specimens

et al. 1989

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Consecutive clinical specimens of Chlamydia trachomatis (1,048) were inoculated in parallel on DEAEdextranand cycloheximide-treated HeLa 229 cells and cycloheximide-treated McCoy cells. HeLa 229 cell culture detected 113 positive specimens, and McCoy cell culture detected 103 positive specimens. This difference is not significant. However, HeLa 229 cell culture yielded significantly more inclusions than McCoy cell culture in the 95 specimens positive in both cell types (P = 0.042). For routine diagnostic purposes, a choice for one of the cell types may be determined by local preferences.

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“…The current accepted methods of isolation of C. trachomatis are generally modifications of the in vitro cell culture method first introduced by Gordon et al (4). The conventional procedure is laborious and cumbersome and requires a lengthy incubation period before and after infection of the McCoy cell monolayers, making it impractical for routine testing of large numbers of specimens (1).…”

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Micro direct inoculation method for the isolation and identification of Chlamydia trachomatis

邓勇¹,

Paul²

1986

J Clin Microbiol

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Current standard procedures for the isolation and identification of Chlamydia trachomatis are laborious. A more rapid method for the isolation and identification of C. trachomatis has been developed. This method features the use of a multiwell microplate to which trypsinized McCoy cells and specimens are added simultaneously, followed by centrifugation. A simplified one-step iodine-glycerol staining procedure is used to detect the chlamydial inclusions. The requirements of the conventional macro method for preseeding cell monolayers and for the use of cumbersome cover slips in glass vials are eliminated. Studies on 345 clinical specimens showed that the micro method is as sensitive as the conventional method but can be performed more conveniently, rapidly, and economically.

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Comparison of methods for cultivation and isolation of Chlamydia trachomatis

Cited by 16 publications

References 18 publications

Enhancement of Chlamydia trachomatis infectious progeny by cultivation of HeLa 229 cells treated with DEAE-dextran and cycloheximide

Enhancement of Chlamydia trachomatis infectious progeny by cultivation of HeLa 229 cells treated with DEAE-dextran and cycloheximide

Comparison of HeLa 229 and McCoy cell cultures for detection of Chlamydia trachomatis in clinical specimens

Micro direct inoculation method for the isolation and identification of Chlamydia trachomatis

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