Declaration: As with all diagnostic tests, a definitive clinical diagnosis should not be based on the result of a single test, but should only be made by the physician after all clinical and laboratory findings have been evaluated.
INTENDED USE
SUMMARY
PRINCIPLE
STORAGE AND STABILITY
WARNING AND PRECAUTIONS
SPECIMEN COLLECTION AND STORAGE
TEST PROCEDURE
INTERPRETATION OF RESULTS
LIMITATIONS OF PROCEDURE
QUALITY CONTROL
SPECIFICITY
REFERENCE
Catalog No. Product Name
ABT-STD-B18 Chlamydia Cassette
INTENDED USE
Accu-Tell ® Chlamydia Antigen Test Cassette is intended for in vitro diagnostic use in the rapid, qualitative detection of Chlamydia trachomatis directly from female endocervical swab, male urethral swab, male urine specimens, and ocular specimens. The test is intended for use as an aid in the diagnosis of Chlamydia infections.
SUMMARY
The genus Chlamydia includes three species: Chlamydia trachomatis, and the recently described Chlamydia pneumoniae, a primarily human pathogen, and Chlamydia psittasi, primarily animal pathogen. Chlamydia trachomatis comprise 15 known serovars, is associated with trachomatis and gentourinary infection, and three serovars are associated with lymphogranuloma venereum (LGV). Chlamydia trachomatis infections are the most common bacterial sexually transmitted diseases. Approximately 4 million new cases occur each year in the United States, primarily cervicitis and nongonococcal urethritis. This organism also causes conjunctivitis, and infant pneumonia. Chlamydia trachomatis infection has both a high prevalence and asymtomatic carriage rate, with frequent serious complications in both women and neonates. Complications of Chlamydia infection in women include cervictis, urethritis, endometritis, pelvic inflammatory diseases (PID) and increased incidence of ectopic pregnancy and infertility. Vertical transmission of the disease during parturition from mother to neonate can result in inclusion conjunctivitis and pneumonia. In men at least 40% of the cases of nongonococcal urethritis are associated with Chlamydia infection and epididymitis. Approximately 70% of women with endocervical infections and up to 50% of men with urethral infections are symthomatic.
Chlamydia psittasi infection is associated with respiratory disease in individuals exposed to infected birds and is not transmitted from human to human. Chlamydia pneumonia, first isolated in 1983, is associated with respiratory infections and pneumonia. Traditionally, Chlamydia infection has been diagnosed by the detection of Chlamydia inclusions in tissue culture cells. Culture method is the most sensitive and specific laboratory method, but it is labor intensive, expensive, long time (2-3 days) and not routinely available in most institutions. Direct tests such as immunofluorescence assay (IFA) requires specialized equipment and a skilled operator to read the result.
PRINCIPLE
The Chlamydia Rapid Test Cassette is a rapid qualitative immunoassay based on the immunochromatographic principle. In the assay procedure, a clinical specimen is obtained and placed into an extraction tube containing Extraction A. After 2 minutes Extraction B is added to the tube. After extraction, 4 drops (approximately 200 μl) of the extracted sample are added to the test cassette sample well. In the test, the membrane was precoated with antigenus-specific lipopolysaccharide (LPS) monoclonal antibody on the test band region and goat anti-mouse antibody on the control band region. During testing, the sample is allowed to react with the colloidal gold particles which have been coated with monoclonal anti-Chlamydia antibody, then moves laterally on the membrane by capillary action. If the sample contains Chlamydia antigen, a colored band with a specific antibody-Chlamydia-antibody-colloidal gold particle complex will form on the membrane in the test band region. If Chlamydia antigen is not present, only a pink line on the control band region appears. To serve as procedural control, a colored band at the control region will always appear regardless the presence of Chlamydia.
STORAGE AND STABILITY
The test kit is to be stored at room temperature (2~ 30°C) in the sealed pouch for the duration of the shelf life. Do not freeze.
WARNING AND PRECAUTIONS
1. For in vitro use only.
2. Do not use kit components after the expiration date. Do not mix kit components from different lots. Do not mix solution bottle caps.
3. Use appropriate precautions in the collection, handling, storage and disposal of the specimens and used kit contents. All specimens, reagents and controls should be handled as if they contain infectious agents. When the assay procedure is complete, dispose the swabs carefully after autoclaving them at 121°C for at least 20 minutes. Alternatively, they can be treated with 0.5% sodium hypochloride (or house-hold bleach) for one hour before disposal.
4. Extraction solution A contains sodium hydroxide (a basic solution); Extraction solution B contains Hydrochloric acid (an acid solution). If either of the solutions contacts the skin or eye, flush with large volumes of water.
5. Positive and negative controls contain sodium azide which may react with lead or copper plumbing to form potentially explosive metal azides. When disposing these solutions, always flush with copious amounts of water to prevent azide buildup. Use only sterile dracon swabs or cytology brushes to obtain endocervical specimens.
6. Do not eat, drink or smoke in the area where specimens and kit reagents are handled. Wear protective clothing such as laboratory coats and disposable gloves while collecting and assaying samples.
7. As with all diagnostic tests, a definitive clinical diagnosis should not be based on the result of a single test, but should only be made by a physician after all clinical and laboratory findings have been evaluated.
SPECIMEN COLLECTION AND STORAGE
The quality of specimen obtained is of extreme importance. Detection of Chlamydia requires a vigoros and thorough collection technique which provides cellular material rather than just body fluids.
For cervical specimens:
1. Use the swab provided with the kit. Alternatively any shafted swabs with rayon or dracon tips may be used.
2. Before specimen collection, remove excess mucus from the endocervical area with a separate swab or cotton ball and discard. The swab should be inserted into the endocervical canal, past the squamocolumnar junction, until most of the tip is no longer visible. This will permit acquisition of columnar or cuboidal epithelial cells which are the main reservoir of Chlamydia organismn. Firmly rotate the swab for 15 – 20 seconds without contamination with exocervical or vaginal cells.
3. Alternatively, endocervical specimens can be collected with a cytology brush (caution: do not use cytology brushes with pregnant patients). After cleaning the exocervix with dracon swab, insert the cytology brush into the endocervical canal, past the squamocolumnar junction. Leave in place two to three seconds, rotate the cytology brush two full turns, withdraw the brush without touching any vaginal surface.
4. Put the swab to the extraction tube, if the test may be run immediately.
For Urethral specimens:
1. Standard wire-shafted fiber-tipped swabs, or cytology brushes (not provided) should be used for urethral specimen collection. Instruct the patients not to urinate at least one hour prior to specimen collection.
2. Insert the swab into the urea about 2-4 cm, rotate for 3-5 seconds and withdraw it, and place it in the extraction tube, if the swab may be tested immediately.
Do not place the swab in any transport device containing medium since transport medium interferes with the assay and viability of the organisms is not required for the assay. If immediate testing is not possible, the patient samples should be placed in a dry transport tube for storage or transport. The swabs may be stored for 4 hours at room temperature (15-30°C) or 24 hours refrigerated (2-8°C). Do not freeze . All specimens should be allowed to reach a room temperature of 15-30°C before testing.
For male urine specimens:
1. Instruct the patient to collect at least 20 to 40 ml of first catch urine in a sterile container (not provided with the kit) without any preservative added. First morning urine specimen is prefered to achieve the highest probability of Chlamydia antigen recovery.
2. If the urine specimens are not to be tested immediately, they can be stored refrigerated (2-8°C ) for 24 hours.
TEST PROCEDURE
I. Specimens and control extraction:
1. Prepare Endocervical or Urethral swab specimens:
• Place a clean Extraction tube in the designated area of the workstation. Add 8 drops of Extraction A to the extraction tube.
• Immerse the patients swab into the Extraction tube and extract 2 minutes at room temperature. During extraction, use a circular motion to roll the swab against the side of the Extraction tube so that the liquid is expressed from the swab and can reabsorbe.
• At the end of the extraction time, add 8 drops of solution B. Squeeze the swab firmly against the tube to expel as much liquid as possible from the swab. Discard the swab following guidelines for handling infectious agents.
• The specimens extracted can retain at room temperature for 60 minutes without affecting the result of the Chlamydia test.
2. Prepare Male urine only:
• Collected urine specimens should be centrifuged in order to collect all particulate matter eventually containing Chlamydia cells. Centrifuge the urine (at least 15 ml) at 10,000 rpm for 10 minutes.
• Carefully discard the supernatant and add 8 drops of Extraction A to the tube and resuspend the pellet by a disposable pipette until the suspension is homogeneous. Extract 2 minutes.
• Transfer the suspension to the extraction tube using a disposable pipette. Add 8 drops of solution B.
3. Prepare Positive and Negative Control (provided on request, only):
• Place a clean Extraction tube in the designated area of the workstation. Add 8 drops of Extraction A to the extraction tube.
• Add two drops of the Positive and Negative Control solution to sterile dracon swabs. Allow the drops to be absorbed into the swab.
• Continue with the assay as described in the Test Procedure using these swabs in place of a patient specimen.
II. Test procedure
• Review the package insert instructions.
• Remove the Chlamydia Antigen Test from the protective pouch and place it on a clean, dry and level surface. Label the device with patient or control identifications.
• Add 3 drops (approximately 150 μl) of extracted sample from the Extraction Tube to the sample well on the test cassette.
• Wait for colored bands to appear. The test should be read within 10 minutes after adding the extract suspension to the sample. Depending on the number of the Chlamydia on the swab, some results may be visible as soon as 1 minute. However, to confirm negative results, the complete reaction time of 15 minutes is required. Do not interpret results after 20 minutes.
INTERPRETATION OF RESULTS
Negative: only one red colored band appears on the control line region. No apparent red-colored band is visible on the test line region. This indicates that no Chlamydia antigen has been detected.
Positive: in addition to the control band, a distinct red colored band also appears on the test line region. This indicates that the specimen contains Chlamydia antigen.
Invalid: when no colored band appears on either region, the test should be voided. This indicates a possible error in performing the test. A new test device should be repeated. Either a fresh specimen may be collected or remaining extraction mixture can be used for this purpose.
LIMITATIONS OF PROCEDURE
The Chlamydia Antigen Test does not specifically differenti-ate C. trachomatis, C. pneumonia or C. psittaci. Detection of Chalmydia is depen-dent on the number of organisms present in the specimen. This may be affected by specimen collection methods and patient factors such as age, history of STD, presence of symptoms, etc. The minimum detection level of this test may vary according to serovar.
QUALITY CONTROL
1. The Chlamydia Antigen Test includes a procedural control. A pink colored band appearing in the control region of the membrane indicates proper performance and reactive reagents.
2. Good laboratory practice includes the use of external control to ensure proper kit performance. Each day of testing, two levels of commercial controls should be tested on the Chlamydia Test. The two levels of controls should consist of a negative control and a positive control containing low level of Chlamydia. The use of the low level positive control will assure that the test has not been adversely affected and detecting Chlamydia at the stated sensitivity of the test system. For this purpose we recommend the use of the Positive and Negative Control solutions supplied in the kit.
SPECIFICITY
The antibody used in Chlamydia Rapid Test Cassette has been shown to detect all 15 Chlamydia serovars. In addition, Chlamydia psittasi and Chlamydia pneumonia strains have been tested with Chlamydia Rapid Test Cassette and gave a positive result.
Cross reactivity with other organisms has been studied using suspensions of 107 CFU/mL. The following organisms were not detected using Chlamydia Rapid Test Cassette:
Acinetobacter
calcoaceticus
Salmonella typhi
Staphylococcusaureus
Neisseria gonorrhoeae
Neisseria catarrhalis
Neisseria meningitidis
Neisseria lactamica
Pseudomonas aeruginosa
Proteus vulgaris
Acinetobacter spp
Candida albicans
Escherichia coli
Gardnerella vaginalis
Streptococcus faecalis
Streptococcus faecium
Trichomonas vaginalis
REFERENCE
1. Grayston, J. T., C.-C. Kuo, S.-P. Wang, and J. Altman. N. Engl. J. Med. 315:161-168. 1986.
2. CDC.Morbid. Mortal. Weekly Rep. 34 (suppl. 3-s):535. 1985.
3. Barnes, R. Clin. Microbiol. Rev. 2:119-136. 1989.
4. Kellogg, J. A. Arch.Pathol. Lab. Med. 113:453-460. 1989.
5. Schachter, J. N. Engl. J. Med. 13:105-114. 1978.
6. Schachter, J. Chlamydiae, p.1045-1053. Manual of Clinical Microbiology, 5th ed. American Society for Microbiology, Washington, D.C. 1991.
7. Schachter, J., and C. R. Dawson. Sex. Transm. Dis. 8:167-174. 1981.
8. Stamm, W. E. Ann. Intern. Med. 108:710-717. 1988.
9. Miller, M. J. Journal Med. Tech. 2(8) : 507 – 512. 1985.
The above information is just for reference. The actual technical specifications are subject to the insert provided with the product.
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