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HelpEpic Test Code BRBST Tickborne Bacterial, PCR and Sequencing, Blood
Necessary Information
Specimen source is required.
Specimen Required
Container/Tube:
Preferred: Lavender top (EDTA)
Acceptable: Royal blue top (EDTA), pink top (EDTA), or sterile vial containing EDTA-derived aliquot
Specimen Volume: 1 mL
Collection Instructions: If not submitting in original vial, mix well before transferring to a sterile vial.
Useful For
Detecting and identifying pathogenic tickborne bacteria infecting normally sterile whole blood
Potential detection of bacteria that cause similar illnesses to tickborne infections
This test should not be used as first tier test. It should only be used when routine testing is negative.
This test is not recommended as a test of cure because nucleic acids may persist for long periods of time after successful treatment.
Reflex Tests
| Test ID | Reporting Name | Available Separately | Always Performed |
|---|---|---|---|
| SPID2 | Specimen Identification by PCR | No, (Bill Only) | No |
Testing Algorithm
For information see Acute Tickborne Disease Algorithm.
Special Instructions
Method Name
16S Ribosomal RNA Gene Polymerase Chain Reaction (PCR) followed by Next Generation Sequencing (NGS)
Reporting Name
Tickborne Bacterial PCR+Sequence, BSpecimen Type
Whole Blood EDTASpecimen Minimum Volume
0.5 mL
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| Whole Blood EDTA | Refrigerated (preferred) | 14 days | |
| Frozen | 14 days |
Reject Due To
All specimens will be evaluated at Mayo Clinic Laboratories for test suitability.Clinical Information
The target population is patients with suspected, but undiagnosed, tickborne bacterial infection involving normally sterile whole blood. Polymerase chain reaction (PCR) amplification of a portion of the 16S ribosomal RNA gene followed by next-generation sequencing of the amplified product can be used to detect tickborne bacterial nucleic acids in such situations, enabling a diagnosis. Ideal specimens are those that specific tickborne PCR tests or blood culture have not resulted in identifiable causative infectious agents. Due to the complexity of this test, the suspected tickborne disease testing algorithm will reflex to this assay only if specific-PCR tests are negative. The test is designed to identify mono-bacterial or poly-bacterial tickborne infections.
Reference Values
No tickborne bacterial DNA detected
Interpretation
A positive broad-range polymerase chain reaction (PCR)/sequencing result indicates that tickborne bacterial nucleic acid was detected.
A negative sequencing result indicates the absence of detectable bacterial nucleic acids in the specimen but does not rule out false-negative results that may occur due to sampling error, sequence variability underlying the primers, the presence of bacterial nucleic acids in quantities less than the limit of detection of the assay, or inhibition of PCR amplification. If testing shows evidence of PCR inhibition, it will be repeated. If inhibition is again detected, the result will be reported as "PCR inhibition present."
Cautions
This test does not detect nonbacterial organisms (eg, viruses, fungi, helminths, protozoa).
False-positive results are theoretically possible if patient specimens are contaminated with bacterial nucleic acids from the environment or patient microbiota (eg, skin microbiota contamination).
This test is validated for whole blood only.
Supportive Data
Fifty-two positive patient specimens were available for accuracy studies and correlated with results of organism-specific polymerase chain reaction (PCR). In addition, 20 negative samples from previous PCR testing were evaluated in verification. Using criteria established in verification, overall sensitivity of the assay is 82.7%, and specificity is 100% compared to single-analyte PCR tests. Sensitivity was lower due to the suboptimal recovery of Borrelia burgdorferi and increased to 97.4% without B burgdorferi included. A comment is added to reports that will reflect the limitations of detecting this organism.
The limit of detection was determined using four different organisms (Enterococcus gallinarum, Pseudomonas aeruginosa, Leptospira interrogans, and Anaplasma phagocytophilum) with an average sensitivity of 27 colony forming units per mL of whole blood for E gallinarum and P aeruginosa, and 63 copies per mL of whole blood for L interrogans and A phagocytophilum.
Specificity was tested using a panel of 10 nucleic acid extracts from viral, fungal, and parasitic organisms. No cross-reactivity to these organisms was observed.
Inclusivity studies were performed by sequencing 56 samples representing diverse types of bacteria (including tickborne bacteria). All bacteria were detected and correctly identified by next-generation sequencing.
Day(s) Performed
Monday through Friday
Report Available
14 to 28 daysSpecimen Retention Time
7 daysPerforming Laboratory
Mayo Clinic Laboratories in Rochester
CPT Code Information
87801-Broad Range Bacterial PCR and Sequencing
87798-Specimen Identification by PCR (if appropriate)