Microbiology - Bordetella Pertussis
Specimens for isolation of Bordetella pertussis and B. parapertussis in suspected cases of whooping cough are accepted from public and private health care providers. PCR screening is available for Bordetella pertussis only. Only symptomatic contacts of diagnosed cases of pertussis are recommended for Bordetella examination, since a carrier state in asymptomatic persons has not been demonstrated as an important source of transmission. Reference cultures are accepted for confirmation of Bordetella pertussis, B.parapertussis and B.bronchiseptica. Consultation and bench training are provided upon request.
Specimen Collection
Nasopharyngeal swabs should be collected as soon as possible after onset of symptoms, and prior to antibiotic treatment. There is a greater likelihood of positive cultures and/or PCR in the first two weeks of symptomatic infection than during later weeks of illness. However, PCR may detect organisms for a prolonged period of time regardless of viability.
A mailer containing materials and instructions necessary for collecting and shipping nasopharyngeal specimens is available from the Laboratory Mailroom by ordering online at https://slphreporting.ncpublichealth.com/labportal. Transport medium in the mailer has a shelf life of two months. Notify the unit before submitting large numbers of specimens. Regan-Lowe Transport Medium (RLTM)and 2ml microcentrifuge tube included in the mailer should be labeled with the patient´s name and accompanied by a completed DHHS form #T806. Please do not place adhesive labels on the microcentrifuge tube. Unlabeled specimens will not be tested. Follow collection instructions included in the mailer. The following additional clinical information should be entered on the back of the form: nature of symptoms, date of onset, immunization history, contact with other cases of whooping cough, any antibiotic therapy prior to specimen collection and other pertinent information.
Note: Specimens received without submitter return address are subject to rejection!
Under certain circumstances: If the mailer is not available and culture is not attempted, nasopharyngeal swab specimens may be used for DFA. Swabs should be moistened lightly in a few drops of sterile distilled water or saline and spread thinly over the central area of a glass slide, re-wetting the swab for each slide. Allow smears to air dry and gently heat fix before packaging (see SHIPMENT OF SPECIMENS) with a completed DHHS form #T806. Preferably four smears should be submitted per patient.
Lab records are computerized and the following data are required: patient name, patient Social Security Number (or assigned number), date of birth, patient Medicaid number (if applicable), submitter Federal Tax Number (Employer Identification Number), submitter return address, specimen collection date and specimen source. Without these data, specimen records cannot be entered into the computer, nor can a report of results be printed. Other data are helpful for epidemiological follow-up and for statistical purposes.
Note: CLIA regulations require the following information on all test requisitions:
- Patient name or identifier
- Name and address of submitting agency
- Test requested
- Date specimen collected
Isolated organisms for identification should be subcultured to appropriate media and incubated until growth is apparent before shipping. Bordet-Gengou or Regan-Lowe Agar is recommended for B.pertussis; blood, chocolate or infusion agar is satisfactory for other bordetellae. Agar slants are preferred, but plates may be used if they are sealed in plastic bags and securely packaged. Growth from culture plates also may be suspended in RLTM for shipment or used to prepare smears for DFA staining.
Specimen Shipment
Specimens should be shipped as soon as possible after collection. Clinical specimens for pertussis culture should be shipped refrigerated in cold RLTM using frozen cold packs provided in the insulated mailer. Nasopharyngeal swabs may be held, if necessary, in refrigerated RLTM up to two days before shipping. Friday shipments are not recommended as specimens should be kept cold.
It is essential for culture specimens to be kept cold after collection and during transit to the Laboratory. Another nasopharyngeal swab for PCR inside the microcentrifuge tube also should be returned in the mailer, along with the completed DHHS T806 form.
Reference cultures may be shipped in a microbiology reference mailer with a completed DHHS form #T806. Plates should be wrapped individually in absorbent cushioning material and securely packaged in a leak-proof, crush-proof container. Label "Pertussis" on the outside of the package. When shipping by U.S. mail, use first-class postage. Be sure to place return address on outside of container, regardless of shipping method.
Reporting Procedure and Interpretation
PCR testing is usually performed twice a week. Results of PCR tests are available on the day of test completion, usually within 3 to 4 working days of receipt of sample, and can be accessed via the secure web page for results: https://slphreporting.ncpublichealth.com/lims/. Culture results are reported within seven days after receipt of the specimen; most positive cultures are evident within four days. Positive PCR and culture results are reported by telephone to the submitter, and to the Women’s and Children’s Health Section, Division of Public Health, for surveillance purposes.
Both culture and PCR examinations may fail to detect B.pertussis. As the disease process may continue for weeks or months after viable organisms no longer remain in the nasopharynx, a negative culture report does not rule out infection, especially if specimens were collected late in the course of illness. PCR testing, however, can detect non-viable organisms and has been found to be more sensitive. Organisms present in low numbers may be difficult to detect by either method. Prior antibiotic therapy, overgrowth of contaminants or failure to keep specimens cold after collection and during transit may result in a negative culture. Cultures performed at the local level using commercial agar plates may be negative due to insufficient moisture in the medium.
PCR is a presumptive test. Discrepant PCR and culture reports may occur. Low numbers of organisms may be detected by PCR, but may be overgrown by normal flora in culture.
Reports are returned only to the submitting agency; the submitter is responsible for sending copies to any other agency. Copies of reports are retained in this Laboratory. The submitting agency is responsible for maintaining reports in the patient´s file.
See also:
- About Microbiology
- Botulism
- Bordetella Pertussis
- Enteric Bacteriology
- Foodborne Illness
- Cholera
- Legionella
- Mycobacteriology
- Mycology
- Neisseria Gonorrhoeae
- Norovirus
- Parasitology
- Special & Atypical Bacteriology
