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HelpTest Code PBORR Lyme Disease, Molecular Detection, PCR, Varies
Reporting Name
Lyme Disease PCRUseful For
Supporting the diagnosis of Lyme disease in conjunction with serologic testing
Specific indications including testing skin biopsies when a rash lesion is not characteristic of erythema migrans, and testing synovial fluid or synovium to support the diagnosis of Lyme arthritis
Performing Laboratory
Mayo Clinic Laboratories in Rochester
Specimen Type
VariesSpecimen Required
Ordering Guidance
This assay does not detect Borrelia miyamotoi. If infection with this organism is suspected, order BMIYB / Borrelia miyamotoi Detection PCR, Blood or BMIYC / Borrelia miyamotoi Detection PCR, Spinal Fluid.
Necessary Information
Specimen source is required.
Specimen 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)
Collection Instructions:
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
Spinal Fluid, Synovial Fluid: 0.3 mL
Tissue: NA
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| Varies | Refrigerated (preferred) | 7 days | |
| Frozen | 7 days |
Reference Values
Negative
Day(s) Performed
June through November: Monday through Saturday
December through May: Monday through Friday
Test Classification
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 US Food and Drug Administration.CPT Code Information
87476
87798 x 2
87999 (if appropriate for government payers)
LOINC Code Information
| Test ID | Test Order Name | Order LOINC Value |
|---|---|---|
| PBORR | Lyme Disease PCR | 94253-2 |
| Result ID | Test Result Name | Result LOINC Value |
|---|---|---|
| SRC71 | Specimen Source | 31208-2 |
| 23635 | B. burgdorferi PCR | 94250-8 |
| 38288 | B. mayonii PCR | 94251-6 |
| 38289 | B. garinii/B. afzelii PCR | 94252-4 |
| 38323 | Lyme CSF Comment | 59464-8 |
Testing Algorithm
The following algorithms are available in Special Instructions:
Special Instructions
Clinical Information
Lyme disease is a multisystem and multistage tick-transmitted infection caused by spirochetal bacteria in the Borrelia burgdorferi sensu lato (Bbsl) complex.(1) Nearly all human infections are caused by 3 Bbsl species; B burgdorferi sensu stricto (hereafter referred to as B burgdorferi) is the primary cause of Lyme disease in North America, while B afzelii and B garinii are the primary causes of Lyme disease in Europe. In 2012, B mayonii has been identified as a less common cause of Lyme disease in the upper Midwestern United States.(2,3) This organism has only been detected in patients with exposure to ticks in Minnesota and Wisconsin and has not been detected in over 10,000 specimens from patients in other states including regions of northeast where Lyme disease is endemic.
Lyme disease is the most commonly reported tick-borne infection in Europe and North America, causing an estimated 300,000 cases in the United States each year, and 85,000 cases in Europe.(4,5) The clinical features of Lyme disease are broad and may be confused with various immune and inflammatory disorders. The classic presenting sign of early localized Lyme disease caused by B burgdorferi is erythema migrans (EM), which occurs in approximately 80% of individuals. Other early signs and symptoms include malaise, headache, fever, lymphadenopathy, and myalgia. Arthritis, neurological disease, and cardiac disease may be later stage manifestations. Erythema migrans has also been seen in patients with B mayonii infection, but diffuse rashes are more commonly reported.(2) The chronic skin condition, acrodermatitis chronicum atrophicans, is also associated with B afzelii infection.
The presence of EM in the appropriate clinical setting is considered diagnostic for Lyme disease and no confirmatory laboratory testing is needed. In the absence of a characteristic EM lesion, serologic testing is the diagnostic method of choice for Lyme disease.(6) However, serology may not be positive until 1 to 2 weeks after onset of symptoms, and may show decreased sensitivity for detection of infection with B mayonii. Therefore, detection of Bbsl DNA using PCR may be a useful adjunct to serologic testing for detection of acute disease. PCR has shown utility for detection of Borrelia DNA from skin biopsies of Lyme-associated rashes and also be used to detect Borrelia DNA from synovial fluid and synovium biopsies. Less commonly, Borrelia DNA can be detected in cerebrospinal fluid.(7) Lyme PCR should always be performed in conjunction with FDA-approved serologic tests, and the results should be correlated with serologic and epidemiologic data and clinical presentation of the patient.(8) The Mayo Clinic Lyme PCR test detects and differentiates the main causes of Lyme disease in North America (B burgdorferi and B mayonii) and Europe (B afzelii and B garinii).(2,7)
Interpretation
A positive result indicates the presence of DNA from Borrelia burgdorferi, B mayonii, B afzelii, or B garinii, the main agents of Lyme disease.
A negative result indicates the absence of detectable target DNA 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
Serologic tests are recommended for diagnosis of Lyme disease. PCR may play an adjunctive role, but may not detect Borrelia burgdorferi DNA from cerebrospinal fluid (CSF) in cases of active or chronic disease. The presence of inhibitory substances may also cause a false-negative result. If clinical features of illness are highly indicative of Lyme neuroborreliosis, serologic testing on CSF is warranted. 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.
Testing of CSF by PCR in patients with suspected Lyme neuroborreliosis should be requested only on patients with positive B 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).
Concurrent infections with multiple tick-borne pathogens, including Ehrlichia muris eauclairensis, Anaplasma phagocytophilum, Babesia microti, and B miyamotoi (a relapsing fever Borrelia) have been reported in United States, and consideration should be given to testing for other pathogens if clinically indicated.
This assay detects most members of the Borrelia burgdorferi sensu lato complex, including B andersonii, B americana, and B bissettii, which have been rarely detected in humans. Detection of DNA from these organisms would be reported as an atypical result and prompt additional laboratory testing to further identify the DNA present. The sensitivity of this assay for detecting these organisms has not been determined.
This assay also detects some members of the B burgdorferi sensu lato (Bbsl) complex that are not considered to be human pathogens, but may be found in ticks and other animals. Therefore, this assay should not be used to test nonhuman specimens.
Supportive Data
The following validation data supports the use of this assay for clinical testing.
Analytical Sensitivity/Limit of Detection (LoD):
The lower LoD is approximately 300 to 1,000 genomic copies/mL in cerebrospinal fluid (CSF), tissue, blood, and synovial fluid.
Accuracy/Diagnostic Sensitivity and Specificity:
Spiking studies of whole organism in fresh tissue, synovial fluid, and CSF (spiked near the approximate LoD) showed 100% recovery.
Analytical Specificity:
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, R typhi, Ehrlichia canis, Babesia microti, Plasmodium falciparum, P vivax, Bartonella henselae, Bartonella quintana, Herpes simplex virus, and Toxoplasma gondii. Relapsing fever borreliae (including Borrelia miyamotoi) are also not detected with this assay.
Precision:
Interassay precision was 100% and intra-assay precision was 100%.
Reference Range:
The reference range for this assay is negative. 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 should not be used to screen asymptomatic patients.
Reportable Range:
This is a qualitative assay, and the results are reported as negative or positive for targeted B burgdorferi, B afzelii, B garinii, or B mayonii.
Clinical Reference
1. Stanek G, Wormser GP, Gray J, Strle F: Lyme borreliosis. Lancet 2012;379(9814):461-473
2. Pritt BS, Mead PS, Johnson, DK, et al: Identification of a novel pathogenic Borrelia species causing Lyme borreliosis with unusually high levels of spirochetemia: a descriptive study. Lancet Infect Dis 2016 May;16(5):556-564
3. Pritt BS, Respicio-Kingry LB, Sloan LM, et al: Borrelia mayonii sp. nov., a member of the Borrelia burgdorferi sensu lato complex, detected in patients and ticks in the upper midwestern United States. Int J Sys Evol Microbiol 2016;66(11):4878-4880
4. Hinckley AF, Connally NP, Meek JI, et al: Lyme disease testing by large commercial laboratories in the United States. Clin Infect Dis 2014;59(5):676-681
5. Lindgren E, Jaenson TGT: Lyme borreliosis in Europe: influences of climate and climate change, epidemiology, ecology and adaptation measures. Copenhagen, Denmark: World Health Organization; 2006
6. Centers for Disease Control and Prevention. Recommendations for test performance and interpretation from the Second National Conference on Serologic Diagnosis of Lyme Disease. MMWR Morb Mortal Wkly Rep 1995;44(31):590-591
7. 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
8. 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
Method Description
Nucleic acid is extracted from clinical specimens using the automated MagNA Pure LC instrument system. The extract is then transferred to individual wells of a 96-well plate 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 (OppA2), which is present at a frequency of 1 copy per organism in all 4 confirmed pathogenic species of the Borrelia burgdorferi sensu lato genogroup (B burgdorferi sensu stricto, B afzelii, B garinii and B mayonii). 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 610, at the 5' end. When the target amplicon is present, the LC-Red 610 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 B burgdorferi sensu stricto from B mayonii, B afzelii, and B garinii, although the melting curve analysis cannot differentiate between B afzelii and B 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)
Report Available
1 to 4 daysSpecimen Retention Time
1 weekReject Due To
| All specimens will be evaluated at Mayo Clinic Laboratories for test suitability. |
NY State Approved
YesMethod Name
Real-Time Polymerase Chain Reaction (PCR)/DNA Probe Hybridization
Forms
If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.