Lyme Disease (Borrelia burgdorferi), Molecular Detection, PCR
NY State Approved Indicates the status of NY State approval and if the test is orderable for NY State clients.
Confirmation of active Lyme disease
Supporting the diagnosis of Lyme arthritis
Testing of cerebrospinal fluid (CSF) by PCR in patients with suspected Lyme neuroborreliosis should be requested only on patients with positive Borrelia burgdorferi antibody in serum confirmed by Western blot assay LYWB / Lyme Disease Antibody, Immunoblot, Serum and with abnormal CSF findings (elevated protein and WBC >10 cells/high-power field).
Testing Algorithm Delineates situation(s) when tests are added to the initial order. This includes reflex and additional tests.
See Acute Tick-Borne Disease Testing Algorithm in Special Instructions.
Special Instructions and Forms Describes specimen collection and preparation information, test algorithms, and other information pertinent to test. Also includes pertinent information and consent forms to be used when requesting a particular test
Real-Time Polymerase Chain Reaction (PCR)/DNA Probe Hybridization
(PCR is utilized pursuant to a license agreement with Roche Molecular Systems, Inc.)
Reporting Name A shorter/abbreviated version of the Published Name for a test; an abbreviated test name
Lyme Disease PCR
Borrelia burgdorferi by PCR
Lyme Disease (PCR)
Lyme Disease, CSF
Lyme Disease (PCR)
Lyme Disease, CSF
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:
Specimen Type: Spinal fluid
Container/Tube: Sterile vial
Specimen Volume: 1 mL
Collection Instructions: Label specimen as spinal fluid.
Specimen Type: Synovial fluid
Container/Tube: Sterile vial
Specimen Volume: 1 mL
Collection Instructions: Label specimen as synovial fluid.
Specimen Type: Tissue (fresh only)
Sources: Skin or synovial biopsy
Container/Tube: Sterile container with normal saline
Specimen Volume: Approximately 4 mm(3)
1. Submit only fresh tissue.
2. Skin biopsies:
a. Wash biopsy site with an antiseptic soap. Thoroughly rinse area with sterile water. Do not use alcohol or iodine preparations. A local anesthetic may be used.
b. Biopsy specimens are best taken by punch biopsy to include full thickness of dermis.
3. Label specimen with source of tissue.
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.
Spinal Fluid or Synovial Fluid: 0.3 mL/Tissue: NA
Green-top (heparin) tube, yellow-top (ACD) tube, blood, or plasma
Specimen Stability Information Provides a description of the temperatures required to transport a specimen to the laboratory. Alternate acceptable temperature(s) are also included.
|Varies||Refrigerated (preferred)||7 days|
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
Lyme disease is a multisystem and multistage infection caused primarily by 3 species of tick-borne spirochetes in the Borrelia burgdorferi sensu lato genogroup. These spirochetes include Borrelia burgdorferi sensu stricto (North America and Western Europe), Borrelia afzelii (Central and Western Europe and Russia), and Borrelia garinii (Europe, Russia, and northern Asia). Endemic areas for Lyme disease in the United States correspond with the distribution of 2 tick species, Ixodes scapularis (Northeastern and Upper Midwestern US) and Ixodes pacificus (West Coast US). In Europe, Ixodes ricinus transmits the spirochete. Lyme disease is the most commonly reported tick-borne infection in Europe and North America (CDC).
Lyme disease exhibits a variety of symptoms that may be confused with immune and inflammatory disorders. Inflammation around the tick bite causes skin lesions. Erythema (chronicum) migrans (ECM), a unique expanding skin lesion with central clearing that has a ring-like appearance, is typically the first stage of the disease. Arthritis, neurological disease, and cardiac disease may be later stage manifestations.
Serology is currently the diagnostic method of choice for Lyme disease. However, serology may not be positive until 2 to 4 weeks after onset of ECM, and direct detection of Borrelia species. Target DNA using PCR may be a useful adjunct to existing diagnostic tests for acute disease. PCR has shown utility for detection of Borrelia DNA from skin biopsies of ECM lesions, as well as DNA from synovial and cerebrospinal fluid in late-stage disease.(1) Borrelia DNA can also, rarely, be detected from blood, but is not the test of choice from this source.
Lyme PCR may be useful for adjunctive testing to support a serologic diagnosis of Lyme disease, and should be performed in conjunction with FDA-approved serologic tests.
PCR results should be correlated with serologic and epidemiologic data and clinical presentation of the patient.
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.
A positive result indicates the presence of DNA from Borrelia burgdorferi, the agent of Lyme disease.
A negative result indicates the absence of detectable DNA from Borrelia burgdorferi in the specimen. Due to the clinical sensitivity limitations of the PCR assay, a negative result does not preclude the presence of the organism or active Lyme disease.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
A negative result does not rule-out Lyme disease, since inhibitory substances may be present in the specimen and the assay has limited clinical sensitivity when testing certain specimen types (eg, cerebrospinal fluid: CSF). If clinical features of illness are highly indicative of Lyme neuroborreliosis, serologic testing on CSF is warranted. Patients with active infection due to Borrelia afzelii or Borrelia garinii may have positive results from this PCR test, which will be reported as atypical gene sequence and prompt additional testing. PCR test results should be used as an aid in diagnosis and not considered diagnostic by themselves. These results should be correlated with serologic and epidemiologic data and clinical presentation of the patient.
Concurrent infections with multiple tick-borne pathogens, including Ehrlichia chaffeensis/Anaplasma phagocytophilum and Babesia microti have been reported in United States, and consideration should be given to testing for other pathogens if clinically indicated.
The following validation data supports the use of this assay for clinical testing.
Accuracy/Diagnostic Sensitivity and Specificity:
Results from this real-time PCR assay on the LightCycler (LC PCR) directed to the plasminogen-binding protein were compared to those generated using conventional PCR (target ospA gene) for synovial fluid (82), whole blood (22), and cerebrospinal fluid (CSF) (85). Using the conventional PCR as the gold standard, the diagnostic sensitivity and specificity for detection of Borrelia burgdorferi was as follows: synovial fluid (98.1% and 100%), whole blood (100% and 100%), and CSF (80% and 100%).
Additional spiking studies of whole organism in fresh tissue, synovial fluid, CSF, and whole blood (spiked near the approximate limit of detection) showed 100% recovery.
Analytical Sensitivity/Limit of Detection (LoD):
The lower LoD is approximately 300 to 1,000 genomic copies/mL in CSF, tissue, synovial fluid, and whole blood.
No PCR signal was obtained from the extracts of 22 bacterial, viral, parasitic, and fungal isolates that can cause symptoms similar to Lyme disease including: Rickettsia rickettsii, Rickettsia typhi, Ehrlichia canis, Babesia microti, Plasmodium falciparum, Plasmodium vivax, Bartonella henselae, Bartonella quintana, Herpes simplex virus, and Toxoplasma gondii.
Inter-assay precision was 100% and intra-assay precision was 100%.
Although the reference range is "negative" for this assay, it may detect low-grade asymptomatic bacteremia from individuals exposed to Lyme-endemic areas. However, this assay is only to be used for patients with a clinical history and symptoms consistent with Lyme, and must be interpreted in the context of serologic tests, which are the gold standard for diagnosis of Lyme disease. This test is not used to screen asymptomatic patients.
This is a qualitative assay, and the results are reported as negative or positive for targeted Borrelia burgdorferi.
Clinical Reference Provides recommendations for further in-depth reading of a clinical nature
1. Nocton JJ, Bloom BJ, Rutledge BJ, et al: Detection of Borrelia burgdorferi DNA by polymerase chain reaction in cerebrospinal fluid with Lyme neuroborreliosis. J Infect Dis 1996;174:623-627
2. CDC: Recommendation for test performance and interpretation. From second national conference on serological diagnosis of lyme disease. MMWR Morb Mortal Wkly Rep 1996;45:481-484
3. Nocton JJ, Dressler F, Rutledge BJ, et al: Detection of Borrelia burgdorferi DNA by polymerase chain reaction in synovial fluid from patients with Lyme arthritis. N Engl J Med 1994;330:229-234
4. Babady NE, Sloan LM, Vetter EA, et al: Percent positive rate of Lyme real-time polymerase chain reaction in blood, cerebrospinal fluid, synovial fluid, and tissue. Diagn Microbiol Infect Dis 2008;62(4):464-466
Method Description Describes how the test is performed and provides a method-specific reference
Nucleic acid is extracted from clinical specimens using the automated MagNA Pure LC instrument system. The extract is then transferred to individual self-contained capillary cuvettes for amplification. The LightCycler is an automated instrument that amplifies and monitors the development of target nucleic acid (amplicon) after each cycle of PCR. The DNA target for PCR assay is the 283-bp plasminogen-binding protein gene (pbp), which is present at a frequency of 1 copy per organism in all 3 pathogenic species of the Borrelia burgdorferi sensu lato genogroup (Borrelia burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii). A specific base pair DNA target sequence is amplified by PCR. The detection of amplicon is based on fluorescence resonance energy transfer (FRET), which utilizes 1 hybridization probe with a donor fluorophore, fluorescein, at the 3' end, and a second hybridization probe with an acceptor fluorophore, LC-Red 640, at the 5' end. When the target amplicon is present, the LC-Red 640 emits a measurable and quantifiable light signal at a specific wavelength. Presence of the specific organism nucleic acid may be confirmed by performing a melting curve analysis of the amplicon. Using features of the melting curve analysis, the assay primers and specific hybridization probes are able to detect and differentiate Borrelia burgdorferi sensu stricto from Borrelia afzelii, and Borrelia garinii. The melting curve analysis of this assay cannot differentiate between Borrelia afzelii and Borrelia garinii. Each assay run can be completed within 60 minutes.(Cockerill FR, Uhl FR: Applications and challenges of real-time PCR for the clinical microbiology laboratory. In Rapid Cycle Real-Time PCR. Edited by U Reischl, C Wittwer, F Cockerill. Springer, NY 2002)
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 Saturday (June through November)
Monday through Friday (December through May)
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.
This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the U.S. Food and Drug Administration.
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.
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|
|23635||Lyme Disease PCR||4991-6|