Test ID BPRPV Bordetella pertussis and Bordetella parapertussis, Molecular Detection, PCR, Varies
Necessary Information
Specimen source is required.
Specimen Required
The high sensitivity of amplification by polymerase chain reaction (PCR) requires the specimen to be processed in an environment in which contamination of the specimen by Bordetella pertussis or Bordetella parapertussis DNA is unlikely.
Submit only 1 of the following specimens:
Preferred:
Specimen Type: Nasopharyngeal swab
Supplies:
Culture Swab - Liquid Stuarts/Single Swab (NP Swab) (T515)
Container/Tube: Rayon swab with an aluminum shaft placed in transport medium such as a green-top nasopharyngeal swab (rayon mini-tip) with Stuart's media (no charcoal), or Stuart's media with charcoal, or Amies media with or without charcoal (Transwab Nasopharyngeal with Charcoal System).
Additional Information:
1. Swab transport containers without charcoal must contain a pledget saturated with either Stuart's or Amies liquid media. Clear semi-solid/solid media is gel and will be rejected.
2. Other swab or media types may be inhibitory to PCR testing and will be rejected.
Acceptable:
Specimen Type: Nasopharyngeal (not throat) aspirate/wash or nasal aspirate/wash
Container/Tube: Sterile container with a screw top cap (no transport media)
Specimen Volume: Entire collection
Useful For
Preferred diagnostic test for the detection of Bordetella pertussis or Bordetella parapertussis
This test is not recommended for screening asymptomatic individuals who may carry B pertussis or parapertussis.
This test is not recommended for follow up of patients previously diagnosed with pertussis (ie, as a test of cure).
Method Name
Polymerase Chain Reaction (PCR)/DNA Probe Hybridization
Reporting Name
Bordetella, PCR, VariesSpecimen Type
VariesSpecimen Minimum Volume
0.5 mL
Specimen Stability Information
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Varies | Refrigerated (preferred) | 7 days | |
Ambient | 7 days | ||
Frozen | 7 days |
Reject Due To
Nose, nasal, or throat swab Calcium alginate or cotton-tipped swab Swab sent in gel transport medium, viral/universal transport medium, or Regan Lowe media ESwab Swabs with solid plastic shaft Dry swab |
Reject |
Clinical Information
Bordetella pertussis is the highly contagious etiological agent of pertussis or whooping cough. Bordetella parapertussis causes a similar, but generally less severe, illness. Despite vaccination efforts, B pertussis remains common in the United States, underscoring the need for effective diagnostic tests. In the United States, pertussis is most common in the late summer months. Pertussis vaccination does not prevent B parapertussis infection, which generally occurs in a younger age group than disease caused by B pertussis. Diagnosis of pertussis is based on having a high clinical index of suspicion for the infection, along with confirmation by laboratory testing. Laboratory testing methods include nucleic acid amplification tests (eg, polymerase chain reaction [PCR]), serology, culture, and direct fluorescent antibody testing. Culture and direct fluorescent antibody testing are limited by low sensitivity, rendering nucleic acid amplification and serology the tests of choice.
The Centers for Disease Control and Prevention recommends PCR testing for patients suspected of having acute pertussis. B pertussis PCR detects roughly twice as many cases as culture. After symptom onset B pertussis DNA can be detected up to 4 weeks or longer (up to 8 weeks in our experience).(1) However, over time, the amount of B pertussis and B parapertussis DNA will diminish, rendering the assay less sensitive. A serologic response to B pertussis is typically mounted within 2 weeks following infection, and therefore, detection of IgG-class antibodies to pertussis toxin, which is only produced by B pertussis, can be a useful adjunct for diagnosis at later stages of illness at a time when the amount of B pertussis may be below the limit of detection of the PCR assay.
Reference Values
Not applicable
Interpretation
A positive result indicates the presence of DNA from Bordetella pertussis or Bordetella parapertussis. In some cases, a patient may test positive for both B pertussis and B parapertussis. Cross-reactivity with Bordetella holmesii and Bordetella bronchiseptica may occur with the B pertussis assay (see Cautions).
A negative result indicates the absence of detectable B pertussis and B parapertussis DNA in the specimen but does not negate the presence of organism or active or recent disease (known inhibition rate of <1%) and may occur due to inhibition of polymerase chain reaction, sequence variability underlying primers and/or probes, or the presence of B pertussis or B parapertussis in quantities less than the limit of detection of the assay. Additionally, patients presenting late after symptom onset may test negative; in such cases, testing for B pertussis antibody, IgG, in serum (BORDG / Bordetella pertussis Antibody, IgG, Serum) may be considered.
Cautions
Cross-reactivity with Bordetella holmesii may occur with the Bordetella pertussis polymerase chain reaction (PCR) assay. The prevalence of B holmesii is relatively low, with positivity in less than 1% of nasopharyngeal swabs.(2) Note: B holmesii has been associated with pertussis-like symptoms.(2)
Cross-reactivity of the B pertussis assay has been demonstrated with a limited number of Bordetella bronchiseptica isolates. The prevalence of the insertion sequence target, IS481, has been reported to be between 1% and 5% in B bronchiseptica isolates.
Some B pertussis acellular vaccines (ie, Pentacel, Daptacel, Adacel) contain PCR detectable DNA. Contamination of specimens with vaccine can cause false-positive B pertussis PCR results. Specimens should not be collected or processed in areas that are exposed to B pertussis vaccine material.
Clinical Reference
1. Theofiles AG, Cunningham SA, Chia N, et al: Pertussis outbreak, southeastern Minnesota, 2012. Mayo Clin Proc. 2014 Oct;89(10):1378-1388
2. Guthrie JL, Robertson AV, Tang P, et al: Novel duplex real-time PCR assay detects Bordetella holmesii in specimens from patients with pertussis-like symptoms in Ontario, Canada. J Clin Microbiol. 2010 Apr;48(4):1435-1437
3. Sloan LM, Hopkins MK, Mitchell PS, et al: Multiplex LightCycler PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis in nasopharyngeal specimens. J Clin Microbiol. 2002 Jan;40(1):96-100
4. Karalius VP, Rucinski SL, Mandrekar JN, Patel R: Bordetella parapertussis outbreak in Southeastern Minnesota and the United States, 2014. Medicine (Baltimore). 2017 May;96(20):e6730
Method Description
The LightCycler instrument platform amplifies and monitors the development of target nucleic acid sequences by fluorescence after each cycle of polymerase chain reaction (PCR). The automated detection of amplified products is based on the fluorescence resonance energy transfer principle. The assay uses the repetitive (50-100 copies) insertion sequence (IS481) found in Bordetella pertussis and the repetitive (35-50 copies) insertion sequence (IS1001) found in Bordetella parapertussis as targets. Detection and differentiation of Bordetella targets is performed through melting curve analysis. The probes were designed to obtain a 10 degree C temperature shift between B pertussis and B parapertussis that is seen in the melting curve analysis. Analysis of the PCR amplification and probe melting curves is accomplished through the use of the LightCycler software.(Sloan LM, Hopkins MK, Mitchell PS, et al: Multiplex LightCycler PCR assay for detection and differentiation of Bordetella pertussis and Bordetella parapertussis in nasopharyngeal specimens. J Clin Microbiol. 2002 Jan;40(1):96-100; van der Zee A, Schellekens JF, Mooi FR. Laboratory diagnosis of pertussis. Clin Microbiol Rev. 2015 Oct;28(4):1005-26. doi: 10.1128/CMR.00031-15)
Day(s) Performed
Monday through Friday, Sunday
Report Available
1 to 3 daysTest Classification
This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US Food and Drug Administration.CPT Code Information
87798 x 2
87798 (if appropriate for government payers)
NY State Approved
YesForms
If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.