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Lab Manual for UCSF Clinical Laboratories

Lab Manual for SFGH

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CHLAMYDIA CULTURE

Item Value
Available Stat? No
Test code CHLI
Performed by? Microbiology (Chlamydia)
Sendout? no
Price range $
In House Availability Specimens may be collected at any time and refrigerated until transported to the lab (within 72 hours of collection). Specimens are processed Monday through Friday in the Chlamydia Lab (Bldg. 30, Room 417)
Principle Chlamydia trachomatis, C. pneumonia and C. psittaci are obligate intracellular parasites. These organisms can be isolated by culture in various cell lines. Appropriate specimens are inoculated into 1 dram shell vials containing McCoy or HEP2 cell monolayers. After 72-96 hour incubation, the cells are stained with iodine or fluorescein conjugated monoclonal antibody and microscopically scanned to look for characteristic chlamydial inclusions. A second passage into a new McCoy or /HEP2 cell monolayer is also performed for each specimen. Final results are generally available 7-10 days after receipt of specimen in the chlamydia lab.

Specimens that cannot be processed by the Hologic Aptima (Gen-Probe) Combo2 (molecular test) should be sent for culture. (See Chlamydia trachomatis Detection by Nucleic Acid Amplification.)
Interpretation The detection of chlamydial inclusions (positive result) is indicative of disease, and treatment should be instituted. A negative result may indicate the absence of chlamydia or may be due to improper specimen collection and/or transport or small numbers of the organism which were not detected. A result of cytopathic effect (CPE) is reported when the McCoy or HEP2 cell monolayer is destroyed because of toxicity due to improper specimen collection, mucus or the presence of bacteria and/or yeast.

Results of cultures which are positive for chlamydia are phoned directly to the clinician. Preliminary results may be obtained at 72-96 hrs. Final reports are generally available within 7 to 10 days after receipt of specimen in Chlamydia Lab.

Container type Universal viral transport medium for viruses and chlamydia is available from Lab Support Services, 415-206-8199. Collection kits include swabs and a tube of transport medium.
Collection Instructions Collect specimens for Chlamydia culture using the universal viral transport medium for viruses and chlamydia available from Lab Support Services, 415-206-8199. Collection kits include swabs and a tube of transport medium. DO NOT collect specimens for culture with Hologic Aptima Combo2 (molecular test) swabs. DO NOT use wooden-shafted swabs as they are toxic to chlamydia.

Clean away mucus or discharge before collecting specimen. Collect specimen with a swab provided in the package. For cervical specimens from non-pregnant women, collection of specimen with cytobrush is preferred. Satisfactory specimens must contain cells. Place the swab in the universal transport medium. Screw cap on transport tube tightly. Specimens should be sent directly to the lab. If a specimen cannot be sent within an hour, IT SHOULD BE REGFRIGERATED. Samples can be kept at 4° C for 72 hr; otherwise, specimens should be frozen at -70° C.
Sample type Acceptable Specimens
Infants: Nasopharyngeal/throat specimens should be collected from the posterior nasopharynx. Endotracheal washings are acceptable. If conjunctivitis is present, then use a swab to collect epithelial cells from the lower eye lid.
Women: Collect cervical, urethral, rectal swabs; PID - biopsies (fallopian tube).
Men: Urethritis - urethral swabs; proctitis – rectal swabs
MSM: Urethral, pharyngeal & rectal swabs.
Adult Respiratory (C. pneumoniae) or Suspected psittacosis: Nasopharyngeal/throat swabs, lung biopsy, BAL and sputum samples.
Conjunctivitis: Adult inclusion conjunctivitis – collect eye swab from the lower lid. Suspected trachoma – collect eye swab from the upper lid.
LGV: If bubo present, fine needle aspirate of the inguinal node. If C. trachomatis is isolated, then a separate request is needed to genotype the organism*.
Non-reactive arthritis (Reiter's Syndrome): urethral, conjunctiva and synovial fluid.

Unacceptable specimens
Chlamydia are obligate intracellular organisms. Thus, columnar epithelial cells are needed to isolate the organism. Urine, discharge/mucus, lesions, abscess, and most body fluids (e.g. amniotic, peritoneal) are toxic to the cell lines. Feces are only acceptable for C. psittaci. Vulva and vaginal** (>12 yrs old) swabs are inappropriate, as chlamydia do not infect these sites. Prostatic fluid and semen are acceptable only if a urethral swab is also submitted.

*Procedure involves sequencing for pmpHgene of LGV. This test is a send out. Thus, Lab Medicine Resident approval must be obtained.
**Vaginal swabs (<12 yrs old) will be processed only if there is suspected sexual abuse.
Special instructions For additional information on specimen appropriateness or specimen handling, call the Microbiology Lab Medicine Resident, 415-206-5699 or pager 415-443-1438, or the Chlamydia Lab, Bldg 30, Rm 417, at 415-824-5117.
Stability Chlamydia are stable in the universal transport medium for up to 72 hours at refrigerated temperature; i.e., 4°C.
Additional information Note: Specimens which are obtained as part of funded research should be submitted with Chlamydia Request forms from Dr. Schachter's laboratory and should be sent directly to Bldg. 30, Room 417.
References
1. Mardh, P-A, Westrom L, Colleen S, and Wolner-Hanssen P. Sampling, specimen handling, and isolation techniques in the diagnosis of chlamydial and other genital infections. Sex. Trans. Dis. 8:280-285. 1981.
2. Papp J, Schachter J, Gaydos C, and Van Der Pol B. Recommendations for the laboratory-based detection of Chlamydia trachomatis and Neisseria gonorrhoeae-2014. MMWR, 63(2): 1-18.
3. Schachter, J and Wyrick, PB. 1994. Culture and isolation of Chlamydia trachomatis. Methods Enzymol. 236:377-390.
4. Scidmore, MA. 2005. Cultivation and Laboratory Maintenance of Chlamydia trachomatis. Current Protocols in Microbiology. 00:A:11A.1:11A.1.1–11A.1.25.


Last Updated 4/2/2015 10:56:33 AM
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