Product Manual

ManualsBrandsQuidel ManualsLyme TestsSofia Lyme FIA Test Kit, 25/kt

Sofia Lyme FIA Page 1 of 19

FOR USE WITH SOFIA ONLY

For in vitro diagnostic use only.

INTENDED USE

The Sofia Lyme FIA employs immunofluorescence for the rapid differential detection of human IgM and IgG

antibodies to Borrelia burgdorferi from serum and plasma specimens from patients suspected of B. burgdorferi

infection. This qualitative test is intended for use as an aid in the diagnosis of Lyme disease. A negative result

does not preclude infection with B. burgdorferi. All positive results for IgM and/or IgG should be further tested

by a corresponding second-tier western blot assay. Test results are to be used in conjunction with information

obtained from the patient’s clinical evaluation and other diagnostic procedures.

SUMMARY AND EXPLANATION

Lyme disease (LD) is the most common tickborne disease in North America and Europe.¹ In the United States,

LD is caused by the bacterium, Borrelia burgdorferi, transmitted through the bite of an infected blacklegged

tick.¹

Patients infected with B. burgdorferi may experience symptoms associated with three stages: early localized

disease, early disseminated disease, and late persistent disease.

The most characteristic symptom of early

localized disease is the appearance of erythema migrants (EM) on the skin, which appears in up to 80% of

cases.

1,3

EM may also be accompanied by flu-like symptoms (headache, abdominal pain, and fatigue) days or

weeks after infection.

In the second stage, early disseminated disease, untreated patients may begin to see

neurological and rheumatological manifestations, and less commonly, dermatological, cardiac, or

ophthalmological manifestations. These symptoms generally appear weeks to months after infection.

If the

disease continues to be left untreated, late persistent disease may also follow months or years later.³ In late

stage disease, patients may see continued progression of manifestations in the joints, heart, skin, and nervous

system.²

Early detection and treatment of LD can help resolve symptoms and prevent progression of the disease.

The

primary means of identifying B. burgdorferi infection is detection of the body’s IgM and IgG antibody response

using immunoassay.³ Detection of IgM antibodies to B. burgdorferi is generally most significant in the earlier

stages of the disease. Conversely, detection of IgG antibodies has proven to be significant for longer periods, as

the antibodies may remain detectable years after infection.

PRINCIPLE OF THE TEST

The Sofia Lyme FIA is an immunofluorescence-based, lateral flow assay for detection of IgM and/or IgG

antibodies to Borrelia burgdorferi in patient specimens. Reagents for the assay are ready-to-use and provided

in the kit.

Summary of content (19 pages)

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FOR USE WITH SOFIA ONLY For in vitro diagnostic use only. INTENDED USE The Sofia Lyme FIA employs immunofluorescence for the rapid differential detection of human IgM and IgG antibodies to Borrelia burgdorferi from serum and plasma specimens from patients suspected of B. burgdorferi infection. This qualitative test is intended for use as an aid in the diagnosis of Lyme disease. A negative result does not preclude infection with B. burgdorferi.
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The assay uses a bidirectional test strip format to detect both IgM and IgG antibodies to B. burgdorferi. One side of the test strip detects IgM antibodies to B. burgdorferi and the other side of the test strip detects IgG antibodies to B. burgdorferi. To perform the test, the patient specimen (30 µL serum/plasma) is added to a pre-filled Reagent Vial. 100 µL of the diluted sample is then dispensed into the round sample well located near the center of the Test Cassette.
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 As the detection reagent is a fluorescent compound, no visible results will form. Sofia must be used for result interpretation.  Testing should be performed in an area with adequate ventilation.  Dispose of containers and unused contents in accordance with Federal, State and Local regulatory requirements.  Wear suitable protective clothing, gloves, and eye/face protection when handling the contents of this kit.  Wash hands thoroughly after handling.
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NOTE: If the Calibration Check does not pass, notify the on-site Supervisor or contact Quidel Technical Support for assistance Monday through Friday from 7:00 a.m. to 5:00 p.m. Pacific Time at 800.874.1517 (in the U.S.); 858.552.1100 (outside the U.S.); Fax: 858.552.7905; technicalsupport@quidel.com (Technical Support); or contact your local distributor. Built-in Procedural Controls The Sofia Lyme FIA contains a built-in procedural control feature.
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2. Following the prompt on the screen, scan the QC Card (located on the assay kit box). 3. Sofia will prompt the user to select the desired mode (WALK AWAY or READ NOW) and then to run the External Controls. A full description of each mode can be found in the “Using Sofia” section of this Package Insert. 4. Use the following procedure to test each of the Control solutions. The Positive Control must be run first, followed by the Negative Control. a.
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cycles. Make sure sample is at room temperature before using in the test, and for samples that have been previously refrigerated or frozen, mix well by fully thawing and inverting the tube 10 times. Performance of the Sofia Lyme FIA has been established with the following tubes: • Serum Separator Tube (RED) • Serum Separator Tube (TIGER) • Lithium Heparin • Sodium Heparin Note: EDTA is NOT compatible with Sofia Lyme FIA.
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4. Fill a calibrated micropipette with 100 μL of diluted sample from the Reagent Vial. 5. Dispense the contents of the micropipette into the round Test Cassette sample well. 6. Proceed to the next section, “Using Sofia,” to complete the test. USING SOFIA WALK AWAY/READ NOW Modes Refer to the Sofia User Manual for operating instructions. Sofia may be set to two different modes (WALK AWAY and READ NOW). The procedures for each mode are described below.
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Run Test 1. Input the User ID using the barcode scanner or manually enter the data using the key pad. NOTE: If you mistakenly scan the incorrect barcode, use the Arrow Buttons on the Sofia key pad to rehighlight the field. Then simply rescan using the correct barcode, and the previous one will be overwritten with the correct barcode. 2. Input Patient ID and/or Order # using the barcode scanner or manually enter the data using the key pad. 3. Press Start Test and the Sofia drawer will automatically open.
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4. Verify that the correct development mode, WALK AWAY or READ NOW, has been selected. Insert the prepared patient Test Cassette into the drawer of Sofia and gently close the drawer. 5. Sofia will start automatically and display the progress as shown in the example below. In WALK AWAY Mode, the test results will be displayed on the screen in approximately 10 minutes. In READ NOW Mode, the test results will be displayed on the screen in approximately 1 minute. See Interpretation of Results section.
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Positive Results: This display shows a valid positive result for IgM antibodies to B. burgdorferi. Interpret to be presumptive of an acute infection to B. b. Test the specimen with IgM Western blot. This display shows a valid positive result for IgG antibodies to B. burgdorferi. Interpret to be presumptive of a past exposure to B. b. Test the specimen with IgG Western blot. This display shows a valid positive result for IgM and IgG antibodies to B. burgdorferi.
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Invalid Results: This display shows an invalid result. If the test is invalid, a new test should be performed starting with Step 1 and a new Test Cassette. LIMITATIONS  Failure to follow the Test Procedure may adversely affect test performance and/or invalidate the test result.  False positive results may occur if samples are read before the minimum 10 minute development time.  Antibody detection methods do not provide definitive results for establishing or ruling out a diagnosis of Lyme disease.
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Lyme disease. The summarized results in Tables 2 and 3 are presented to provide additional information about the performance of the Sofia Lyme FIA compared to Western Blot with a blinded, characterized serum panel. This does not imply an endorsement of the assay by the CDC. Table 2 Sofia Lyme FIA tested with CDC Panel – IgM Results Clinical Status Negative Controls Early Lyme EM Positive Late Lyme Sofia Lyme IgM % Agreement Neg with clinical Status 157 82.
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Table 5 Lyme IgM Results for Sofia Compared to Predicate Assay Sofia IgM Predicate IgM Category n Pos Neg % Sens 95% CI Pos Equiv Neg %Sens1 95% CI Acute, < 1 month, 64 39 25 60.9% 48.7-72.0% 28 8 28 56.3% 44.1-67.7% with EM Acute, 1-2 months, 4 3 1 75.0% 28.9-96.6% 2 0 2 50.0% 15.0-85.0% with EM Convalescent, 3-12 15 11 4 73.3% 47.6-89.5% 6 2 7 53.3% 30.1-75.2% months, with EM Late Lyme (>1 yr), 12 8 4 66.7% 38.8-86.5% 7 3 2 83.3% 54.0-96.5% Neuro or Arthritic All Categories 95 61 34 64.2% 54.2-73.
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Table 7 Lyme IgM Results for Sofia Compared to Predicate Assay IgM Method Comparison (Prospective Study): 1st Tier PPA and NPA Analysis Predicate Lyme IgM Positive Equivocal Negative % Agreement Sofia IgM 95% CI Positive 37 20 67 PPA** = 56.4% (57 / 101) 46.7% - 65.7% Negative 7 37* 510 NPA = 88.4% (510 / 577) 85.5% - 90.
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Table 9 IgM Second Tier Testing IgM Method Comparison (Prospective Study): 2nd Tier PPA Analysis 1st Tier PPA 56.4% Tier 1 + or ± IgM WB + IgM WB (95% CI) (46.7-65.7%) 2nd Tier PPA 89.2% Predicate IgM 101* 37 63 (95% CI) (74.7-96.3%) Sofia IgM 124** 45 76 Predicate + Sofia IgM 57* 33 23 *11 sample had insufficient volume to be tested on WB **3 samples had insufficient volume to be tested on WB IgM Method Comparison (Retrospective Study): 2nd Tier PPA Analysis 1st Tier PPA 90.
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Table 11 Sofia Lyme FIA Precision – Within Run IgM Positive IgM or IgG Sample IgM % Positivity IgG Positive IgG % Positivity Run 1 Run 2 Total (n=96) Run 1 Run 2 Total (n=96) Negative 0/48 0/48 0.0% 0/48 0/48 0.0% High Negative (C5) 3/48 5/48 8.3% 2/48 5/48 7.3% Low Positive (C95) 48/48 48/48 100% 48/48 46/48 97.
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(RED), Serum Separator Tube (TIGER), Lithium Heparin, and Sodium Heparin. EDTA is not compatible with Sofia Lyme FIA. Interfering Substances A study was performed to assess potential interfering substances with the Sofia Lyme FIA. The substances and concentrations tested are described in Table 14, there was no interference or cross-reactivity results when tested with the Sofia Lyme FIA.
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Disease State Diagnosis Rheumatoid Factor Rickettsia Rocky Mountain Spotted Fever Syphilis # of Samples IgM Positive Results IgG Positive Results 10 0/10 0/10 3 0/3 0/3 10 0/10 0/10 28 8/28 4/28 ASSISTANCE If you have any questions regarding the use of this product or if you want to report a test system problem, please call Quidel Technical Support at 800.874.1517 (in the U.S.) or 858.552.1100, Monday through Friday, from 7:00 a.m. to 5:00 p.m., Pacific Standard Time. If outside of the U.S.
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Sofia Lyme FIA Page 19 of 19