Korean J Clin Microbiol.  2010 Jun;13(2):73-78. 10.5145/KJCM.2010.13.2.73.

Comparison of Collagen-coated Polyethylene Terephthalate Disc Plate and Shell Vial Culture Method for the Isolation of Chlamydophila pneumoniae

• Kim MH
• Lee WK

• KMID: 1461640
• DOI: http://doi.org/10.5145/KJCM.2010.13.2.73

Abstract

BACKGROUND
Chlamydophila pneumoniae is one of the major respiratory infectious pathogens and can be accurately diagnosed by cell culturing. The author performed this study to compare the usefulness of the collagen-coated polyethylene terephthalate (PET) disc culture method and that of the shell vial method.
METHODS
Twenty-nine sputums and 17 blood specimens collected from 46 patients for C. pneumoniae culture were inoculated into HeLa-229 cell monolayers cultured in shell vials and polyester plates. After incubation, they were stained using the indirect immunofluorescent method with genus-specific FITC-conjugated anti-chlamydia antibody. When both results were inconsistent, microimmunofluorescence results were used.
RESULTS
HeLa-229 cells successfully formed monolayers in shell vials and collagen-coated PET plates in all cases. Positive inclusion bodies in HeLa-229 cells of shell vials and PET plates for C. pneumoniae culture were similarly stained with the indirect immunofluorescent method. Both methods showed consistent results with 20 positive and 22 negative cases. The total agreement between the PET plate and shell vial was excellent (91.3%, k=0.826).
CONCLUSION
The collagen-coated PET disc culture method showed highly consistent results with that of the shell vial method, and no technical differences were experienced between the two methods. Therefore, the author concluded that the shell vial method could be replaced by the PET plate method for detection of C. pneumoniae.

Keyword

Collagen-coated PET disc plate; Shell vial; Chlamydophila pneumoniae; Cell culture

MeSH Terms

Cell Culture Techniques
Chlamydial Pneumonia
Chlamydophila
Chlamydophila pneumoniae
Humans
Inclusion Bodies
Phthalic Acids
Pneumonia
Polyesters
Polyethylene
Polyethylene Terephthalates
Sputum
Phthalic Acids
Polyesters
Polyethylene
Polyethylene Terephthalates

Figure

• 

  Fig. 1. (A) Shell vial and round cover glass (12 mm). The cover-glass is located onto the bottom of the shell vial. (B) Collagen-coated polyethylene terephthalate (PET) plate (4-well) and PET disc (15 mm). The PET discs are located onto the bottoms of the wells.

• 

  Fig. 2. A monolayer of HeLa-229 cells with the unoccupied surface and cell boundary and the condensed nuclear chromatin using inverted phase-contrast microscopy (×400). The HeLa-229 cells were cultivated at 37°C with 5% CO2 in shell vial (A) and collagen-coated polyethylene terephthalate (PET) plate (B).

• 

  Fig. 3. Positive inclusion bodies in shell vial (A) and collagen-coated polyethylene terephthalate (PET) plate (B) stained with indirect immunofluorescent method using genus-specific FITC-conjugated anti-chlamydia antibody (×200).

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