Test ID: BPRP
Bordetella pertussis and Bordetella parapertussis, Molecular Detection, PCR
NY State Approved 
Indicates the status of NY State approval and if the test is orderable for NY State clients.
Useful For Suggests clinical disorders or settings where the test may be helpful
Preferred diagnostic test for the detection of Bordetella pertussis and/or Bordetella parapertussis
Method Name A short description of the method used to perform the test
Polymerase Chain Reaction (PCR)/DNA Probe Hybridization
(PCR is utilized pursuant to a license agreement with Roche Molecular Systems, Inc.)
Includes DNA extraction from specimen, PCR amplification, and hybridization
Reporting Name A shorter/abbreviated version of the Published Name for a test; an abbreviated test name
Aliases Lists additional common names for a test, as an aid in searching
Bordetella Pertussis Detection by Polymerase Chain Reaction (PCR)
Parapertussis
PCR (Polymerase Chain Reaction)
Pertussis
Polymerase Chain Reaction (PCR)
Whooping Cough
Bordetella parapertussis by PCR
Bordetella parapertussis by Rapid PCR
Specimen Type Describes the specimen type needed for testing
Specimen Required Defines the optimal specimen. This field describes the type of specimen required to perform the test and the preferred volume to complete testing. The volume allows automated processing, fastest throughput and, when indicated, repeat or reflex testing.
Forms: If not ordering electronically, submit a Microbiology Request Form (Supply T244) with the specimen.
Specimen source is required.
Submit only 1 of the following specimens:
Preferred:
Specimen Type: Nasopharyngeal swab
Container/Tube: Rayon swab with an aluminum shaft placed in transport medium such as Stuart's with charcoal or Amies with charcoal (Transwab Nasopharyngeal with Charcoal System [Supply T286])
Additional Information: 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
Specimen Volume: Entire collection
Specimen Minimum Volume Defines the amount of specimen required to perform an assay once, including instrument and container dead space. Submitting the minimum specimen volume makes it impossible to repeat the test or perform confirmatory or perform reflex testing. In some situations, a minimum specimen volume may result in a QNS (quantity not sufficient) result, requiring a second specimen to be collected.
Reject Due To Identifies specimen types and conditions that may cause the specimen to be rejected
| Hemolysis | NA |
| Lipemia | NA |
| Icterus | NA |
| Other | 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; specimen in Stuart's or Amies medium without charcoal |
Specimen Stability Information Provides a description of the temperatures required to transport a specimen to the laboratory. Alternate acceptable temperature(s) are also included.
| Specimen Type | Temperature | Time |
|---|---|---|
| Varies | Refrigerated (preferred) | 7 days |
| Ambient | 7 days |
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
Pertussis is an infectious respiratory disease caused by the Bordetella pertussis bacterium. Pertussis afflicts unvaccinated individuals, and those in whom immunity has waned. Infants are at particular risk for severe disease and complications. Adults present with a prolonged chronic cough illness. Bordetella parapertussis may cause a similar illness, especially in children; however, the symptoms are less severe and of shorter duration than those associated with Bordetella pertussis.
The wide prevalence of pertussis and its changing epidemiology highlight the need for sensitive and rapid methods for diagnostic testing. Clinical diagnosis of pertussis is complicated because the characteristic cough (whoop) is rarely seen in infant and adult patients.
Several diagnostic methods are available, but many lack sensitivity and/or require repeat testing or extended incubation times for test results. The reference method has traditionally been direct culture of the organism from nasopharyngeal secretions. However, due to the fastidious nature of the organism, recovery by culture is difficult, as the organism is susceptible to environmental exposure (change in temperature and drying) and has specific growth requirements. Direct fluorescent antibody testing of material collected by a nasopharyngeal swab lacks sensitivity.(1) PCR is the Mayo-recommended test for detection of Bordetella pertussis and parapertussis in patients suspected of having active, untreated pertussis. PCR is preferred over culture because it is faster and demonstrates improved sensitivity
Additional information regarding pertussis PCR testing has been published by the CDC and can be found at URL: http://www.cdc.gov/pertussis/clinical/diagnostic-testing/diagnosis-pcr-bestpractices.html
Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.
Not applicable
Interpretation Provides information to assist in interpretation of the test results
A positive result indicates the presence of DNA from Bordetella pertussis or Bordetella parapertussis. In some cases, a patient may test positive for both Bordetella pertussis and Bordetella parapertussis. Cross-reactivity with Bordetella holmesii and Bordetella bronchiseptica may occur with the Bordetella pertussis assay (see Cautions).
A negative result indicates the absence of detectable Bordetella pertussis or Bordetella 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 PCR, sequence variability underlying primers and/or probes, or the presence of Bordetella pertussis or Bordetella parapertussis in quantities less than the limit of detection of the assay.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
Cross-reactivity with Bordetella holmesii may occur with the Bordetella pertussis PCR assay. The prevalence of Bordetella holmesii is relatively low, with positivity in <1% of nasopharyngeal swabs.(2) Bordetella holmesii has, however, been associated with pertussis-like symptoms.(2)
Cross-reactivity of the Bordetella 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 Bordetella bronchiseptica isolates.
This assay is not recommended for screening asymptomatic individuals who may carry Bordetella pertussis or parapertussis
This assay is not recommended for follow up of patients previously diagnosed with pertussis (ie, as a test of cure).
Supportive Data
Due to recently mandated FDA regulations concerning the sale of analyte specific reagents (ASR), Roche no longer produces Bordetella reagents that were used in our previous assay.(3) In response to this, we developed a rapid real-time multiplex reaction LightCycler PCR assay to detect and differentiate Bordetella pertussis and Bordetella parapertussis (LightCycler BORD-T) in nasopharyngeal swabs/washings. We compared its performance to the previous LightCycler ASR PCR system (LightCycler BORD ASR), which used the Roche ASR. Similar to the LightCycler BORD ASR, the LightCycler BORD-T assay targets the multicopy insertion gene sequences IS481 and IS1001 of Bordetella pertussis and parapertussis, respectively. Results of the LightCycler BORD-T assay were compared to results of the LightCycler BORD ASR assay on 374 nasopharyngeal swabs and washings submitted for Bordetella testing. Fifty-four specimens were positive (48 Bordetella pertussis and 6 Bordetella parapertussis) and 314 specimens were negative by both PCR assays. Five nasopharyngeal specimens were positive for Bordetella pertussis or Bordetella parapertussis by the LightCycler BORD-T and negative by the LightCycler BORD ASR. One nasopharyngeal specimen was positive for Bordetella pertussis by the LightCycler BORD ASR but negative by the LightCycler BORD-T assay. The LightCycler BORD-T assay demonstrated 98% (368/374) agreement with the BORD ASR. Thirty respiratory microorganisms (pathogenic and non-pathogenic), including 4 different Bordetella species, were tested with the LightCycler BORD-T. Bordetella pertussis and Bordetella parapertussis were positive as expected and Bordetella holmesii was detected at the same melt temperature as the Bordetella pertussis IS481 gene target. Bordetella holmesii cannot be distinguished from Bordetella pertussis by the LightCycler BORD-T assay. All other results were negative. The analytical sensitivity of the LightCycler BORD-T was 1 target/mcL for nasopharyngeal swabs and 10 targets/mcL for nasopharyngeal wash/aspirates.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. She RC, Billetdeaux E, Phansalkar AR, et al: Limited applicability of direct fluorescent-antibody testing for Bordetella sp. and Legionella sp. specimens for the clinical microbiology laboratory. J Clin Microbiol 2007;45:2212-2214
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;48:1435-1437
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;40:96-100
4. Dragsted DM, Dohn B, Madsen J, et al: Comparison of culture and PCR for detection of Bordetella pertussis and Bordetella parapertussis under routine laboratory conditions. J Med Microbiol 2004;53:749-754
5. Antila M, He Q, de Jong C, et al: Bordetella holmesii DNA is not detected in nasopharyngeal swabs from Finnish and Dutch patients with suspected pertussis. J Med Microbiol 2006;55:1043-1051
Method Description Describes how the test is performed and provides a method-specific reference
The LightCycler instrument amplifies (multiplex) and monitors the development of target nucleic acid sequences by fluorescence after each cycle of PCR. This is an automated PCR system that can detect amplicon development through stringent air-controlled temperature cycling and cuvettes in approximately 60 minutes. The detection of amplified products is based on the fluorescence resonance energy transfer (FRET) 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 degrees C temperature shift between Bordetella pertussis and Bordetella 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;40:96-100)
Day(s) and Time(s) Test Performed Outlines the days and times the test is performed. This field reflects the day and time the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time required before the test is performed. Some tests are listed as continuously performed, which means assays are performed several times during the day.
Monday through Friday; 10 a.m., Sunday; 10 a.m.
Analytic Time Defines the amount of time it takes the laboratory to setup and perform the test. This is defined in number of days. The shortest interval of time expressed is "same day/1 day," which means the results may be available the same day that the sample is received in the testing laboratory. One day means results are available 1 day after the sample is received in the laboratory.
Maximum Laboratory Time Defines the maximum time from specimen receipt at Mayo Medical Laboratories until the release of the test result
Specimen Retention Time Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded
Performing Laboratory Location The location of the laboratory that performs the test
Test Classification Provides information regarding the medical device classification for laboratory test kits and reagents. Tests may be classified as cleared or approved by the US Food and Drug Administration (FDA) and used per manufacturer's instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR), Investigation Use Only (IUO) product, or a Research Use Only (RUO) product.
CPT Code Information Provides guidance in determining the appropriate Current Procedural Terminology (CPT) code(s) information for each test or profile. The listed CPT codes reflect Mayo Medical Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.
87801
LOINC® Code Information Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the result codes returned for this test or profile.
| Result ID | Reporting Name | LOINC Code |
|---|---|---|
| SRC54 | Specimen source | 31208-2 |
| 34994 | Bordetella pertussis PCR | 43913-3 |
| 34995 | Bordetella parapertussis PCR | 42588-4 |
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