Test Catalog

Test Id : TKPNL

Tick-Borne Panel, Molecular Detection, PCR, Blood

Useful For
Suggests clinical disorders or settings where the test may be helpful

Evaluation of patients with suspected human monocytic ehrlichiosis, human granulocytic anaplasmosis, babesiosis, or Borrelia miyamotoi infection

 

Evaluation of patients with a history of, or suspected, tick exposure who are presenting with fever, myalgia, headache, nausea, and other nonspecific symptoms

Profile Information
A profile is a group of laboratory tests that are ordered and performed together under a single Mayo Test ID. Profile information lists the test performed, inclusive of the test fee, when a profile is ordered and includes reporting names and individual availability.

Test Id Reporting Name Available Separately Always Performed
LBAB Babesia species, PCR, B Yes Yes
EHRL Ehrlichia/Anaplasma PCR, B Yes Yes
BMIYB Borrelia miyamotoi Detection PCR, B Yes Yes

Testing Algorithm
Delineates situations 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
Library of PDFs including pertinent information and forms related to the test

Method Name
A short description of the method used to perform the test

Real-Time Polymerase Chain Reaction (PCR)/DNA Probe Hybridization

NY State Available
Indicates the status of NY State approval and if the test is orderable for NY State clients.

Yes

Reporting Name
Lists a shorter or abbreviated version of the Published Name for a test

Tick-Borne DNA Panel, B

Aliases
Lists additional common names for a test, as an aid in searching

Anaplasma phagocytophilum

Babesia divergens

Babesia duncani

Babesia microti

Babesia MO-1

Babesiosis

Borrelia miyamotoi

E. chaffeensis

Ehrlichia chaffeensis

Ehrlichia ewingii

Ehrlichia muris-like (EML)

Ehrlichia/Anaplasma

EML (Ehrlichia muris-like)

HA (Human Anaplasmosis)

HE (Human Ehrlichiosis)

HGA (Human Granulocytic Anaplasmosis)

HGE (Human Granulocytic Ehrlichiosis)

HME (Human Monocytic Ehrlichiosis)

Human Anaplasmosis (HA)

Human Ehrlichiosis (HE)

Human Granulocytic Anaplasmosis (HGA)

Human Granulocytic Ehrlichiosis (HGE)

Human Monocytic Ehrlichiosis (HME)

Tick Borne

Tick Borne Diseases

Ehlichia muris eauclairensis

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

See Acute Tick-Borne Disease Testing Algorithm in Special Instructions.

Specimen Type
Describes the specimen type validated for testing

Whole Blood EDTA

Specimen Required
Defines the optimal specimen required to perform the test and the preferred volume to complete testing

Container/Tube: Lavender top (EDTA)

Specimen Volume: 1 mL

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Forms

Specimen Minimum Volume
Defines the amount of sample necessary to provide a clinically relevant result as determined by the Testing Laboratory

0.3 mL

Reject Due To
Identifies specimen types and conditions that may cause the specimen to be rejected

Gross hemolysis Reject
Gross lipemia Reject

Specimen Stability Information
Provides a description of the temperatures required to transport a specimen to the performing laboratory, alternate acceptable temperatures are also included

Specimen Type Temperature Time Special Container
Whole Blood EDTA Refrigerated (preferred) 7 days

Useful For
Suggests clinical disorders or settings where the test may be helpful

Evaluation of patients with suspected human monocytic ehrlichiosis, human granulocytic anaplasmosis, babesiosis, or Borrelia miyamotoi infection

 

Evaluation of patients with a history of, or suspected, tick exposure who are presenting with fever, myalgia, headache, nausea, and other nonspecific symptoms

Testing Algorithm
Delineates situations when tests are added to the initial order. This includes reflex and additional tests.

See Acute Tick-Borne Disease Testing Algorithm in Special Instructions.

Clinical Information
Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test

Ticks are the primary vectors of infectious diseases in North America, and rank second only to mosquitoes in disease transmission worldwide. In the United States, tick-borne diseases include Lyme disease, Rocky Mountain spotted fever, human monocytic ehrlichiosis, human granulocytic anaplasmosis, babesiosis, tularemia, relapsing fever, Colorado tick fever, and Borrelia miyamotoi infection.(1) Several of these diseases are transmitted by the same tick, and coinfections are occasionally seen. In particular, Ixodes species ticks are capable to transmitting the causative agents of Lyme disease (B burgdorferi and B mayonii), anaplasmosis (Anaplasma phagocytophilum), and babesiosis (Babesia species). These diseases are prevalent throughout the northeastern and upper Midwestern states and parts of the Pacific Northwest.

 

Symptoms of the various tick-vectored diseases range from mild to life-threatening. Early symptoms, which include fever, aches, and malaise, do not aid in distinguishing the various diseases. Because early treatment can minimize or eliminate the risk of severe disease, early detection is essential, yet patients may not have developed distinctive symptoms to help in the differential diagnosis. A rapid tick-borne PCR panel can assist in identifying the pathogen, allowing treatment to be initiated.

 

While Lyme disease due to B burgdorferi is best detected through 2-tiered serologic testing, acute ehrlichiosis, anaplasmosis, babesiosis, and B miyamotoi infection are best detected using molecular amplification assays. This tick-borne panel offers sensitive, specific, and rapid detection of the agents that cause these 4 diseases.

 

For information on the specific diseases, see the individual test IDs.

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.

Babesia species, MOLECULAR DETECTION, PCR

Negative

 

Ehrlichia/Anaplasma, MOLECULAR DETECTION, PCR

Negative

 

Borrelia miyamotoi, MOLECULAR DETECTION, PCR

Negative

Interpretation
Provides information to assist in interpretation of the test results

Borrelia miyamotoi: 

A positive result indicates the presence of B miyamotoi DNA and is consistent with active or recent infection. While positive results are highly specific indicators of disease, they should be correlated with symptoms and clinical findings of tick-borne relapsing fever.

 

Ehrlichia/Anaplasma: 

Positive results indicate presence of specific DNA from Ehrlichia chaffeensis, E ewingii, E muris eauclairensis, or A phagocytophilum and support the diagnosis of ehrlichiosis or anaplasmosis.

 

Negative results indicate absence of detectable DNA from any of these 4 pathogens in specimens, but it does not exclude the presence of the organism or active or recent disease.

 

Since DNA of E ewingii is indistinguishable from that of E canis by this rapid PCR assay, a positive result for E ewingii/canis indicates the presence of DNA from either of these 2 organisms.

 

Babesia: 

A positive result indicates the presence of Babesia species DNA and is consistent with active or recent infection. While positive results are highly specific indicators of disease, they should be correlated with blood smear microscopy, serological results and clinical findings.

 

A negative result indicates absence of detectable DNA from Babesia species in the specimen, but does not always rule-out ongoing babesiosis in a seropositive person, since the parasitemia may be present at a very low level or may be sporadic.

 

Other tests to consider in the evaluation of a patient presenting with an acute febrile illness following tick exposure include serologic tests for Lyme disease (B burgdorferi), and molecular detection (PCR) for ehrlichiosis/anaplasmosis. For patients who are past the acute stage of infection, serologic tests for these organisms should be ordered prior to PCR testing.

Cautions
Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances

This panel does not detect Borrelia burgdorferi or B mayonii, the causative agents of Lyme disease in the United States. While Lyme PCR (PBORB / Lyme Disease [Borrelia burgdorferi], Molecular Detection, PCR, Blood) can be useful for detecting acute infection with B mayonii, this organism has a limited geographic distribution (upper Midwestern United States) and is therefore not included in this panel. Serology is the preferred method for detection of B burgdorferi.  

 

For more information, see the individual test IDs.

Supportive Data

Borrelia miyamotoi:

The following assay verification data supports the use of this assay for clinical testing.

 

Accuracy/Diagnostic Sensitivity and Specificity: Clinical Samples:

-62 clinical EDTA blood specimens received in the clinical laboratory for Ehrlichia/Anaplasma PCR were tested using the Borrelia miyamotoi PCR assay. Results were compared to the MDH 16S rRNA TaqMan assay.

-In addition, 2 retrospectively identified B miyamotoi-positive specimens were confirmed by the B miyamotoi PCR assay and the MDH TaqMan assay.

 

Spiking studies:

-To supplement the clinical specimens, negative whole blood and cerebrospinal fluid (CSF) specimens were spiked with genomic or plasmid DNA of B miyamotoi near the limit of detection and tested in a blinded fashion. The sensitivity of the PCR assay was 100% and the specificity with spiked specimens was 100%. The samples were extracted and tested in a blinded fashion.

 

Species Inclusivity:

-4 strains of B miyamotoi were detected by the PCR assay.

-3 other Borrelia species that cause human tick-borne relapsing fever (B hermsii, B parkeri, and B turicatae) were also are detected at a melting temperature below that of B miyamotoi. Thus, related Borrelia species may be detected and differentiated with this assay.

-16 different strains from the B burgdorferi sensu lato group were not detected with this PCR assay.

 

Analytical Sensitivity/Limit of Detection (LoD):

-The LoD is 2,800 target copies/mL of whole blood or CSF.

 

Analytical Specificity:

-No PCR signal was obtained from the extracts of 31 bacterial, viral, parasitic, and fungal isolates from similar organisms or from organisms commonly found in the specimens tested.

 

Precision:

-Interassay precision was 100% and intra-assay precision was 100%.

 

Reference Range:

-The reference range of this assay is negative. This assay is designed to detect only species of clinical significance and is to be used for patients with a clinical history and symptoms consistent with tick-borne relapsing fever. It should not be used to screen healthy patients.

 

Reportable Range:

-This is a qualitative assay, and the results are reported as negative or positive for Borrelia miyamotoi DNA.

 

Ehrlichia/Anaplasma:

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) were compared to those generated using conventional PCR assay for A phagocytophilum on 127 unique, archived whole blood specimens (26 positive and 99 negative specimens by conventional PCR). Using the conventional PCR as the gold standard, the diagnostic sensitivity and specificity for detection of A phagocytophilum were 100%. In addition, 12 known E chaffeensis isolates and 2 E ewingii isolates (reference strains) were tested by the LC PCR and were positive. 

 

Supplemental Data (Spiking Studies): 

To supplement the above data, 30 negative whole blood samples were spiked with A phagocytophilum positive control plasmid at the LoD (10 copies/microL). The 30 spiked specimens were run in a blinded manner along with 30 negative (nonspiked) specimens; 100% of the spiked specimens were positive, and 100% of the nonspiked specimens were negative.

 

Analytical Sensitivity/LoD:

The lower LoD of this assay for each of the species in EDTA blood is as follows:

-A phagocytophilum=approximately 10 targets per microliter

-E chaffeensis=approximately 5 targets per microliter

-E muris eauclairensis- =approximately 100 targets per microliter

-E ewingii/canis=approximately 10 targets per microliter

 

Analytical Specificity:

No PCR signal was obtained from extracts of the following organisms: herpes simplex virus, Epstein-Barr virus, Staphylococcus aureus, S epidermidis, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa, Bartonella henselae, B quintana, Rickettsia typhi, R rickettsii, Toxoplasma gondii, Babesia microti MN, B microti ATCC 53899, Borellia burgdorferi ATCC 51990, Ehrlichia risticii ATCC VR-986, and Anaplasma marginale. Positive results were obtained from nucleic extracts of 2 Ehrlichia canis strains (patient strain and ATCC CRL-10390 strain), with a melting temperature (Tm) of 49.5 degrees C (indistinguishable from Ehrlichia ewingii). A positive melting peak was also noted with Ehrlichia muris (ATCC VR-1411), but the Tm (55.24 degrees C) was easily distinguished from the Tm of the target organisms.

 

Precision: 

Interassay precision was 97% and intra-assay precision was 96%.

 

Reference Range: 

Fifty whole blood specimens from normal donors were tested and found to be negative for targeted or detectable Ehrlichia and Anaplasma species.

 

Reportable Range: 

This is a qualitative assay, and results are reported as either negative or positive for targeted Ehrlichia/Anaplasma species (positive for A phagocytophilum, E chaffeensis, E muris eauclairensis , or Ehrlichia ewingii).

 

Babesia:

The following validation data supports the use of this assay for clinical testing.

 

Accuracy/Diagnostic Sensitivity and Specificity:

-96 whole blood specimens were tested by this real-time PCR assay and another real-time PCR assay. Concordance was 99%.

 

Analytical Sensitivity/LoD:

The LoD established using whole organism spiked into specimen matrix (whole blood) is as follows:

-Babesia microti, ATCC PRA 99: 2,670 target copies/mL

-Babesia duncani ATCC PRA 302: 1,540 target copies/mL

-Babesia MO-1 positive patient DNA: 10,700 target copies/mL

-Babesia divergens-positive patient DNA: 5,270 target copies/mL

 

Serial 10-fold dilutions of microscopy-positive specimens were also tested in a blinded fashion using conventional thick and thin blood films and the Mayo Babesia species PCR. The PCR was able to consistently detect two 10-fold dilutions lower than using microscopy.

 

Analytical Specificity:

No cross-reactivity was noted using a panel of 34 bacteria, viruses, parasites, and fungi were detected by the Babesia species PCR.

                         

Precision:  

Interassay and intra-assay precision was 100% precision.

 

Reference Range:  

The reference range is negative. This was confirmed by testing 93 blood specimens from asymptomatic individuals for the presence of Babesia species by the Babesia species PCR assay. All 93 specimens were negative.

 

Reportable Range:  

This test is a qualitative assay, and results are reported as positive or negative for Babesia species (B microti, B duncani, B divergens, and B MO-1).

Clinical Reference
Recommendations for in-depth reading of a clinical nature

Caulfield AJ, Pritt BS: Lyme Disease Coinfections in the United States. Clin Lab Med 2015;35(4):827-846

Special Instructions
Library of PDFs including pertinent information and forms related to the test

Method Description
Describes how the test is performed and provides a method-specific reference

Borrelia miyamotoi:

The assay is performed on the Roche LightCycler (LC) 2.0 instrument, following DNA extraction on the Roche MagNA Pure. The LC 2.0 instrument amplifies and monitors the development of target nucleic acid (amplicon) after each cycle of PCR.

 

The DNA target for this PCR assay is a gene encoding the glycerophosphodiester phosphodiesterase (glpQ) gene specific to Borrelia species in the relapsing fever group. This gene is not found in Borrelia species that cause Lyme disease.

 

The 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 is confirmed by performing a melting temperature analysis of the amplicon; the presence or absence of a melting peak in the appropriate temperature range is used to determine if a specimen is positive or negative.

 

Ehrlichia/Anaplasma:

Nucleic acid is extracted from the pathogens in blood using the automated MagNA Pure LC 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 groEL, the open reading frame gene segment of the heat-shock protein operon (groEL), which is present at a frequency of 1 copy per organism in pathogenic species of Anaplasma and Ehrlichia. 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 a 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 among A phagocytophilum, Ehrlichiosis chaffeensis, E muris eauclairensis, and E ewingii/canis. Due to close proximity of the melting curves of E ewingii and E canis, this assay cannot distinguish between these 2 organisms.

 

Babesia:

Nucleic acid is extracted from EDTA whole blood using the automated MagNA Pure bead-based system (Roche Molecular Systems). 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 a gene encoding the nuclear small subunit ribosomal RNA (SS-rDNA).

 

This assay consists of 2 forward primers, 1 reverse primer, and 2 probes, which are specific for the Babesia species target DNA. The specific base pair DNA target sequence is first amplified by PCR using the target-specific primers. Amplicon is then detected during melting curve analysis using fluorescence resonance energy transfer (FRET) probes, 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. Fluorescence is produced when the 2 probes anneal to the target sequence in close proximity to one another. The LC-Red 640 then emits a measurable and quantifiable light signal at a specific wavelength.(Cockerill FR, Uhl FR: Applications and challenges of real-time PCR for the clinical microbiology laboratory. In

PDF Report
Indicates whether the report includes an additional document with charts, images or other enriched information

No

Day(s) Performed
Outlines the days the test is performed. This field reflects the day that the sample must be in the testing laboratory to begin the testing process and includes any specimen preparation and processing time before the test is performed. Some tests are listed as continuously performed, which means that assays are performed multiple times during the day.

Monday through Saturday

Report Available
The interval of time (receipt of sample at Mayo Clinic Laboratories to results available) taking into account standard setup days and weekends. The first day is the time that it typically takes for a result to be available. The last day is the time it might take, accounting for any necessary repeated testing.

Same day/1 to 4 days

Specimen Retention Time
Outlines the length of time after testing that a specimen is kept in the laboratory before it is discarded

1 week

Performing Laboratory Location
Indicates the location of the laboratory that performs the test

Rochester

Fees
Several factors determine the fee charged to perform a test. Contact your U.S. or International Regional Manager for information about establishing a fee schedule or to learn more about resources to optimize test selection.

  • Authorized users can sign in to Test Prices for detailed fee information.
  • Clients without access to Test Prices can contact Customer Service 24 hours a day, seven days a week.
  • Prospective clients should contact their Regional Manager. For assistance, contact Customer Service.

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 instructions, or as products that do not undergo full FDA review and approval, and are then labeled as an Analyte Specific Reagent (ASR) product.

See Individual Test IDs

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 Clinic Laboratories interpretation of CPT coding requirements. It is the responsibility of each laboratory to determine correct CPT codes to use for billing.

CPT codes are provided by the performing laboratory.

87798 x 8

LOINC® Information
Provides guidance in determining the Logical Observation Identifiers Names and Codes (LOINC) values for the order and results codes of this test. LOINC values are provided by the performing laboratory.

Test Id Test Order Name Order LOINC Value
TKPNL Tick-Borne DNA Panel, B In Process
Result Id Test Result Name Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
84319 Anaplasma phagocytophilum 87558-3
23663 Ehrlichia chaffeensis 87559-1
23664 Ehrlichia ewingii/canis 87560-9
32010 Ehrlichia muris eauclairensis 87561-7
36114 Babesia microti 88452-8
36115 Babesia duncani 88451-0
36116 Babesia divergens/MO-1 88450-2
64970 B. miyamotoi PCR, B 82475-5

Test Setup Resources

Setup Files
Test setup information contains test file definition details to support order and result interfacing between Mayo Clinic Laboratories and your Laboratory Information System.

Excel | Pdf

Sample Reports
Normal and Abnormal sample reports are provided as references for report appearance.

Normal Reports | Abnormal Reports

SI Sample Reports
International System (SI) of Unit reports are provided for a limited number of tests. These reports are intended for international account use and are only available through MayoLINK accounts that have been defined to receive them.

SI Normal Reports | SI Abnormal Reports