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Test ID SLYME Lyme Antibody Modified 2-Tier with Reflex, Serum

Specimen Required

Supplies: Sarstedt Aliquot Tube, 5 mL (T914)

Collection Container/Tube:

Preferred: Serum gel

Acceptable: Red top

Submission Container/Tube: Plastic vial

Specimen Volume: 0.6 mL

Collection Instructions: Centrifuge and aliquot serum into a plastic vial.

Useful For

Diagnosis of Lyme disease


This test should not be used as a screening procedure for the general population.

Testing Algorithm

If the Lyme antibody result is positive or equivocal, then confirmation by whole cell sonicate will be performed at an additional charge.


For more information see Acute Tick-Borne Disease Testing Algorithm.

Method Name

Enzyme-Linked Immunosorbent Assay (ELISA)

Reporting Name

Lyme Ab Modified 2-Tier w/Reflex, S

Specimen Type


Specimen Minimum Volume

0.5 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
Serum Refrigerated (preferred) 10 days
  Frozen  30 days

Reject Due To

Gross hemolysis Reject
Gross lipemia Reject
Gross icterus Reject
Heat-inactivated specimen Reject

Clinical Information

Lyme disease (LD) is caused by infection with a member of the Borrelia burgdorferi sensu lato complex, which includes B burgdorferi sensu stricto (herein referred to as B burgdorferi), Borrelia afzelii, and Borrelia garinii. Among these species, B burgdorferi is the most frequent cause of LD in North America. These tick-borne spirochetes are transmitted to humans through the bite of Ixodes species ticks. Endemic areas for LD 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).


Transmission of LD-associated Borrelia requires at least 36 hours of tick attachment. Approximately 80% of infected individuals will develop a unique expanding skin lesion with a central zone of clearing, referred to as erythema migrans (EM; stage 1). In the absence of treatment, patients may progress to early disseminated disease (stage 2), which is characterized by neurologic manifestations (eg, meningitis, cranial neuropathy, radiculoneuropathy) and is often associated with B garinii infection. Patients with late LD often present with intermittent or persistent arthralgia, most often associated with B burgdorferi infection, or with acrodermatitis chronica atrophicans (ACA), typically due to infection with B afzelii.


Diagnosis of LD is currently based on either the standard or modified 2-tiered serologic testing algorithm (STTTA or MTTTA, respectively). For the STTTA, see LYME / Lyme Disease Serology, Serum.


The MTTTA starts with an initial enzyme immunoassay (EIA) screen for detection of total antibodies against the Borrelia Vlse/pepC10 proteins. Samples that are screen positive or equivocal by this first tier EIA are subsequently reflexed for supplemental assessment using 2 separate EIAs for detection of IgM and IgG antibodies against B burgdorferi whole cell sonicate material.


Importantly, while serologic assessment for LD may be negative in the early weeks following infection, over 90% of patients with later stages of infection are seropositive by serology, which remains the diagnostic method of choice for this disease.

Reference Values


Reference values apply to all ages.



Negative for antibodies to the Borrelia (Borreliella) species causing Lyme disease. Negative results may occur in patients who have been recently infected (≤14 days).  If recent infection is suspected, repeat testing on a new sample collected in 7 to 14 days is recommended.



Not diagnostic. Supplemental testing in accordance with the modified 2-tiered testing algorithm for Lyme disease has been ordered by reflex.



Not diagnostic. Supplemental testing in accordance with the modified 2-tiered testing algorithm for Lyme disease has been ordered by reflex.


A negative result does not exclude the possibility of infection with Lyme disease causing Borrelia species. Patients in the early stages of Lyme disease and those who have been treated with antibiotics may not exhibit detectable antibody titers. Patients with clinical history, signs, or symptoms suggestive of Lyme disease should be retested in 2 to 4 weeks if the initial test result is negative.


A positive result is not definitive evidence of infection with Borrelia burgdorferi. It is possible that other disease conditions may produce artifactual reactivity in the assay (eg, infectious mononucleosis, syphilis). All samples with equivocal or positive results should be tested using a second-tier method, including either immunoblot testing or enzyme immunoassay (EIA) methods for IgM- and IgG-class antibodies in accordance with Centers for Disease Control and Prevention and the Association of State and Territorial Public Health Laboratory Directors (CDC/ASTPHLD) recommendations.


Patients infected with other members of the B burgdorferi sensu lato complex, including Borrelia garinii, Borrelia afzelii, and Borrelia mayonii will be detected by this assay; however, they cannot be differentiated.


This test should not be performed as a screening procedure for the general population. The predictive value of a positive or negative result depends on the prevalence of analyte (antibodies present to VlsE1 and pepC10 antigens) in a specific population. Testing should only be performed when clinical evidence suggests the diagnosis of Borrelia infection or related etiological conditions observed by the physician.


Lyme serology should not be used for treatment monitoring as IgG can remain for years post-resolution of infection. Instead, monitoring resolution of symptoms in response to treatment is recommended.

Clinical Reference

1. Theel ES. The past, present and (possible) future of serologic testing for Lyme disease. J Clin Microbiol. 2016;54(5):1191-1196. doi:10.1128/JCM.03394-15

2. Dattwyler RJ. Lyme borreliosis: An overview of clinical manifestations. Lab Med. 1990;21:290-292

3. Schwan TG, Burgdorfer W, Rosa PA. Borrelia. In: Murray PR, ed. Manual of Clinical Microbiology. 7th ed. ASM Press; 1999:746-758

4. Centers for Disease Control and Prevention (CDC). 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

Method Description

The first-tier Lyme disease screening enzyme-linked immunosorbent assay (ELISA) used is the Zeus ELISA Borrelia VlsE1/pepC10 IgG/IgM test system. The Zeus ELISA Borrelia VlsE1/pepC10 IgG/IgM test system is designed to detect IgG- and IgM-class antibodies (not differentiated by the assay in the final result) in human sera to VlsE1 and pepC10 antigens. Diluted test sera are incubated in antigen coated microwells. Any antigen-specific antibody in the sample will bind to the immobilized antigen. The plate is washed to remove unbound antibody and other serum components. Peroxidase conjugated goat antihuman IgG and IgM are added to the wells and the plate is incubated. The conjugate will react with IgG and IgM antibodies immobilized on the plate. The wells are washed to remove unreacted conjugate. The microwells containing immobilized peroxidase conjugate are incubated with peroxidase substrate solution. Hydrolysis of the substrate by peroxidase produces a color change.  After a period of time the reaction is stopped, and the color intensity of the solution is measured photometrically. The color intensity of the solution depends upon the antibody concentration in the original test sample.(Package insert: Borrelia VlsE1/pepC10 IgG/IgM Test System. Zeus Scientific, Inc; Rev 05/21/2021)

Day(s) Performed

Monday through Friday

Report Available

Same day/1 to 4 days

Test Classification

This test has been cleared, approved, or is exempt by the US Food and Drug Administration and is used per manufacturer's instructions. Performance characteristics were verified by Mayo Clinic in a manner consistent with CLIA requirements.

CPT Code Information


86617 x2 (if appropriate)

LOINC Code Information

Test ID Test Order Name Order LOINC Value
SLYME Lyme Ab Modified 2-Tier w/Reflex, S 83081-0


Result ID Test Result Name Result LOINC Value
SLYME Lyme Ab Modified 2-Tier w/Reflex, S 83081-0

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
TLYME Lyme IgM/IgG, WCS, EIA, S Yes No


If not ordering electronically, complete, print, and send Infectious Disease Serology Test Request (T916) with the specimen.