ImmuView S pneumoniae and L pneumophila Urinary Antigen Test
Device Facts
| Record ID | K191184 |
|---|---|
| Device Name | ImmuView S pneumoniae and L pneumophila Urinary Antigen Test |
| Applicant | Ssi Diagnostica A/S |
| Product Code | MJH · Microbiology |
| Decision Date | Mar 5, 2020 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.3300 |
| Device Class | Class 2 |
Indications for Use
The ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test is an in vitro, rapid, lateral flow test, also known as a lateral flow immunochromatographic assay, intended for the qualitative detection of Streptococus pneumoniae and Legionella pneumophila antigens in urine specimens with symptoms of pneumonia. The assay is intended to aid in diagnosis of S. pneumoniae and L. pneumophila serogroup 1 infections. The assay is further intended to aid in the diagnosis of S. pneumoniae infection of S. pneumoniae antigen in cerebrospinal fluid (CSF). Results from the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test should be interpreted in conjunction with the patient's clinical evaluation and other diagnostic methods.
Device Story
Rapid lateral flow immunochromatographic assay for qualitative detection of S. pneumoniae and L. pneumophila (serogroup 1) antigens. Input: human urine or CSF samples. Principle: lateral flow; rabbit polyclonal antibodies capture target antigens; visual readout of colored lines on test strip. Used in clinical laboratories by professionals. Output: visual presence/absence of test lines indicating infection. Results interpreted alongside clinical evaluation to aid diagnosis of pneumococcal pneumonia or Legionnaires' disease. Enables rapid confirmation to guide antibiotic therapy. Benefits: timely diagnosis, improved patient prognosis.
Clinical Evidence
Clinical performance evaluated via retrospective and prospective studies. Retrospective urine study (N=100 S. pneumoniae, N=98 L. pneumophila) showed S. pneumoniae sensitivity 78% (95% CI 69-85%) and specificity 98.1% (95% CI 95.4-99.3%); L. pneumophila sensitivity 87.8% (95% CI 79.8-92.9%). Prospective study (N=306) showed S. pneumoniae positive agreement 96.0% and L. pneumophila positive agreement 100%. CSF study (N=183) showed S. pneumoniae sensitivity 92.9% and specificity 96.0%. Reproducibility study (N=1072) demonstrated 99.6% overall agreement.
Technological Characteristics
Lateral flow immunochromatographic assay. Materials: test strip with rabbit polyclonal capture antibodies. Energy source: none (passive capillary flow). Dimensions/form factor: test strip/cassette. Connectivity: standalone. Sterilization: not specified. Software: none.
Indications for Use
Indicated for qualitative detection of S. pneumoniae and L. pneumophila serogroup 1 antigens in urine specimens from patients with symptoms of pneumonia, and S. pneumoniae antigen in CSF. For prescription use only by clinical laboratory professionals.
Regulatory Classification
Identification
Haemophilus spp. serological reagents are devices that consist of antigens and antisera, including antisera conjugated with a fluorescent dye, that are used in serological tests to identify Haemophilus spp. directly from clinical specimens or tissue culture isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by bacteria belonging to the genus Haemophilus and provides epidemiological information on diseases cause by these microorganisms. Diseases most often caused by Haemophilus spp. include pneumonia, pharyngitis, sinusitis, vaginitis, chancroid venereal disease, and a contagious form of conjunctivitis (inflammation of eyelid membranes).
Special Controls
*Classification.* Class II (special controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 866.9.
Predicate Devices
Related Devices
- K163273 — TRU Legionella · Meridian Bioscience, Inc. · Feb 21, 2017
- K991726 — BINAX NOWR STREPTOCOCCUS PNEUMONIAE URINARY ANTIGEN TEST (22 TEST KIT),(12 TEST KIT), BINAX NOWR STREPTOCOCCUS PNEUMONI · Binax, Inc. · Aug 27, 1999
- K061496 — SAS LEGIONELLA TEST, SAS LEGIONELLA DIPSTICK TEST · Sa Scientific , Ltd. · Jun 28, 2007
Submission Summary (Full Text)
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FDA
U.S. FOOD & DRUG
ADMINISTRATION
# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
ASSAY ONLY
## I Background Information:
A 510(k) Number
K191184
B Applicant
SSI Diagnostica A/S
C Proprietary and Established Names
ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test
S. pneumoniae and L. pneumophila Urinary Antigen Test
D Regulatory Information
| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| MJH | Class II | 21 CFR 866.3300 - Haemophilus Spp. Serological Reagents | MI - Microbiology |
## II Submission/Device Overview:
A Purpose for Submission:
To obtain a substantial equivalence determination for the detection S. pneumoniae and L. pneumophila antigens in human urine and S. pneumoniae and S. pneumoniae antigens in human CSF.
B Measurand:
S. pneumoniae and L. pneumophila antigens or whole organism
Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.gov
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C Type of Test:
Qualitative lateral flow assay
III Intended Use/Indications for Use:
A Intended Use(s):
The ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test is an in vitro, rapid, lateral flow test, also known as a lateral flow immunochromatographic assay, intended for the qualitative detection of Streptococcus pneumoniae and Legionella pneumophila antigens in urine specimens from patients with symptoms of pneumonia. The assay is intended to aid in diagnosis of S. pneumoniae and L. pneumophila serogroup 1 infections. The assay is further intended to aid in the diagnosis of S. pneumoniae infections by detection of S. pneumoniae antigen in cerebrospinal fluid (CSF). Results from the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test should be interpreted in conjunction with the patient's clinical evaluation and other diagnostic methods.
B Indication(s) for Use:
Same as Intended Use(s)
C Special Conditions for Use Statement(s):
Rx Only
D Special Instrument Requirements:
None
IV Device/System Characteristics:
A Device Description:
The ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test is a lateral flow test for the qualitative detection of S. pneumoniae in human urine and CSF samples and L. pneumophila (primarily serogroup 1) antigens in human urine samples. The test is an aid in diagnosis of pneumococcal pneumonia caused by S. pneumoniae or Legionella pneumonia (Legionnaires' disease), in conjunction with culture and other clinical findings.
B Principle of Operation:
The assay is performed by adding three (3) large free-falling drops (120 μL) of patient urine or CSF are diluted with two (2) large free-falling drops (90 μL) of buffer reagent in a tube. The test strips are introduced into each tube containing either patient samples or controls. After fifteen (15) minutes of sample incubation on the absorbing pad, the test is removed, read and the visual results interpreted. Colored beads bind to the captured antigens, causing the development of red and blue lines respectively. When there are no bacterial antigens present, there are no red/blue
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lines in the test area. As the sample migrates through the membrane a purple/gray line develops in the control (C) area, which consists of goat anti-rabbit antibodies. This built-in procedural control provides evidence that the test was performed properly, and the sample and reagents have migrated through the device strip. If the control line does not appear or if any of the lines are not fully developed the test is invalid.
V Substantial Equivalence Information:
A Predicate Device Name(s):
Binax NOW Legionella Urinary Antigen Test
B Predicate 510(k) Number(s):
K070522
C Comparison with Predicate(s):
| Device & Predicate Device(s): | K191184 | K070522 |
| --- | --- | --- |
| Device Trade Name | ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test | Binax NOW Legionella Urinary Antigen Test |
| General Device Characteristic Similarities | | |
| Intended Use/ Indications For Use | The ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test is an in vitro lateral flow test, also known as a lateral flow immunochromatographic assay, intended for the qualitative detection of Streptococcus pneumoniae and Legionella pneumophila antigen in urine specimens from patients with symptoms of pneumonia. The test is intended to aid in diagnosis of either S. pneumoniae or L. pneumophila serogroup 1. The assay is further intended to aid in the diagnosis of S. pneumoniae infections by detection of S. pneumoniae antigen in cerebrospinal fluid (CSF). Results from the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen should be interpreted in conjunction with the | The BinaxNOW Legionella Urinary Antigen Test is an in vitro rapid immuno-chromatographic assay for the qualitative detection of Legionella pneumophila serogroup 1 antigen in urine specimens from patients with symptoms of pneumonia. It is intended to aid in the presumptive diagnosis of Legionella infection (Legionnaires Disease) caused by L. pneumophila serogroup 1 in conjunction with culture and other methods. |
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| | patient’s clinical evaluation and other diagnostic methods. | |
| --- | --- | --- |
| Measurement Principle | Rapid Lateral flow test | same |
| Read Method | Visual | same |
| Capture antibody | Rabbit polyclonal | same |
| Assay interpretation | Qualitative | same |
| Read time | 15 min | same |
| General Device Characteristic Differences | | |
| Sample type | Urine and CSF | Urine |
| Measured analyte | S. pneumoniae and L. pneumophila | L. pneumophila |
| Sample type | Urine, Urine preserved and CSF | Urine |
VI Standards/Guidance Documents Referenced:
CLSI EP07-A2 Interference Testing in Clinical Chemistry; Approved Guideline-Second Edition, November 2005
CLSI EP12-A2 User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline—Second Edition
CLSI EP17-A2 Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline
CLSI EP25-A Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline, September 2009
FDA Guidance Document- Benefit-Risk Factors to Consider When Determining Substantial Equivalence in Premarket Notifications (510(k)) with Different Technological Characteristics
Guidance for Industry and Food and Drug Administration Staff. Document issued on September 25, 2018.
VII Performance Characteristics (if/when applicable):
A Analytical Performance:
1. Precision/Reproducibility:
Reproducibility of the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test was determined using a 12-member masked specimen panel, consisting of six urine
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specimens, three CSF specimens, and three controls. Of the six urine specimens, two were negative specimens, two were *L. pneumophila* moderate positive specimens, (3 - 4x higher than the C95), one was a *S. pneumoniae* low positive specimen, (1 - 2 x LoD), and one was a *S. pneumoniae* moderate positive specimen (3 - 4x higher than the C95). Of the three CSF specimens tested, one was a true negative specimen, one was a *S. pneumoniae* low positive specimen, (1 - 2 x LoD), and one was a *S. pneumoniae* moderate positive specimen (3 - 4x higher than the C95). Urine and CSF samples were spiked using a known concentration of *S. pneumoniae* or *L. pneumophila* purified antigen to achieve the desired concentration. Testing was performed at two independent laboratories and on-site at SSI. Samples were tested twice a day in triplicate over a five-day period by multiple technicians at each site using multiple kit lots, with a positive control included with each individual test and a negative control tested with each sample panel of masked specimens. Concordance with the expected result for all true negative samples was 99.7% (354/355). For urine, concordance for *L. pneumophila* panel members was 99.4% (356/358) and for *S. pneumoniae* panel members 100% (180/180). For *S. pneumoniae* CSF panel members, concordance was 99.4% (178/179). The test had an invalid rate of 0.74% (1072/1080). The results for reproducibility testing were acceptable.
2. Linearity:
Not Applicable.
3. Analytical Specificity/Interference:
Cross Reactivity: Urine
The ImmuView *S. pneumoniae* and *L. pneumophila* Urine Antigen Test was evaluated for cross-reactivity with the organisms listed in Table 1 below. *S. pneumoniae* and *L. pneumophila* purified antigen was spiked into clinically negative urine at 2-3 x LoD. The organisms listed below were spiked at concentration of >10⁸ CFU/mL and viruses at a range from 10³.³ to 10⁸.²⁵ TCID₅₀ units per 0.2 mL.
Table 1. Organisms tested for cross-reactivity
| Acinetobacter spp.(4) | Lactobacillus rhamnosus |
| --- | --- |
| Bacillus subtilis | Lactobacillus sp. |
| Bordetella pertussis | Listeria monocytogenes |
| Moraxella catarrhalis | Morganella morganii |
| Candida albicans (4) | Moraxella osloensis |
| Citrobacter freundii | Mycoplasma genitalium |
| Cornyebacterium sp. | Neisseria gonorrhoeae (3) |
| Cornyebacterium urealyticum | Neisseria lactamica |
| Enterobacter cloacae (3) | Neisseria meningitidis |
| Escherichia coli (10) | Proteus mirabilis (2) |
| Enterococcus faecalis (7) | Proteus vulgaris |
| Enterococcus faecium | Pseudomonas aeruginosa (4) |
| Enterococcus durans | Pseudomonas stutzeri |
| Gardnerella vaginalis | Pseudomonas spp. (2) |
| Hemophilus influenzae type a-f and non caps (11) | Salmonella bredeney |
| Haemophilus parainfluenzae | Salmonella Thompson |
| Adenovirus 2, | Salmonella typhimurium |
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| Chlamydophila pneumoniae (2) | Serratia marcescens |
| --- | --- |
| Chlamydia trachomatis | Staphylococcus epidermidis |
| Cytomegalovirus | Salmonella glostrup |
| Enterovirus D68 | Streptococcus mutans (2) |
| Herpes Simplex 1,2 | Streptococcus parasanguis |
| Influenza A (H1N1 and H3N2) virus | Streptococcus sanguinis |
| Influenza B Virus | Streptococcus aureus (6) |
| Parainfluenza virus 1,2,3 (3) | Streptococcus epidermidis (5) |
| Respiratory Syncytial Virus A | Streptococcus saprophyticus (3) |
| Klebsiella oxytoca (2) | Stenotrophomonas maltophilia |
| Klebsiella pneumoniae (3) | Streptococcus gr. A, B, C, F, L and G (16) |
| Legionella pneumophila sg3 | Streptococcus mitis |
| Lactobacillus catenaforme | |
## Cross Reactivity: CSF
ImmuView S. pneumoniae and L. pneumophila Urine Antigen Test was evaluated for cross-reactivity with the organisms listed in Table 2 below. The organisms were tested with and without S. pneumoniae purified antigen spiked into clinically negative CSF at 2-3 x LoD. The organisms listed below were spiked at concentration of >10⁸ CFU/mL.
Table 2. Organisms used for Cross-reactivity testing in CSF.
| E. coli (5) | Staphylococcus aureus |
| --- | --- |
| Hemophilus influenza type a-f and non caps (7) | Streptococcus Gr A |
| Listeria monocytogenes | Streptococcus agalactiae (GBS) serogroup g Ia, Ib, II, III (4) |
| Measles | Streptococcus mitis |
| Neisseria meningitidis Gr. B, D and W135 (3) | |
No significant cross reactivity was observed during the study.
## Interfering substances: Urine:
ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test was evaluated for interfering substances with the substances and concentrations listed in Table 3. Testing was conducted using negative urine and negative urine spiked with both S. pneumoniae and L. pneumophila antigen at 2-3x LoD. None of the substances were observed to interfere with the performance of the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen test.
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Table 3 Interfering substances evaluated in urine
| Agent | Concen. | Agent | Concen. |
| --- | --- | --- | --- |
| Acetaminophen | 0.1mg/mL | Leucocytes | >250 cells/mL |
| Acetylsalicyclic acid | 0.1mg/mL | Miconazole | 5% |
| Amantadine | 0.03mg/mL | Mix (pH, whole blood, protein and glucose) (H) | |
| Amoxicillin | 0.075mg/mL | Mix (pH, whole blood, protein and glucose) (M) | |
| Amphotericin B | 0.22mg/mL | Mix (pH, whole blood, protein and glucose) (L) | |
| Antihistamine | 0.22mg/mL | Mucin | 0.086mg/mL |
| Ascorbic acid (C-Vitamin) | 1mg/mL | Oseltamivir (Tamiflu) | 0.03mg/mL |
| Augmentin (Amoxicillin-Clavulanate) | 0.22mg/mL | Oxalic acid | 0.01% |
| Azithromycin | 0.012mg/mL | pH (acidic) | 4 |
| Beet root | 20% | pH (neutral) | 7 |
| Beet root | 1.17% | pH (basic) | 9 |
| Beet root | 0.01% | Plasma | 90% |
| Bilirubin | 0.2mg/mL | Plasma | 50% |
| Bromhexin/cough drops/couch syrup | 0.22mg/mL | Plasma | 10% |
| Caffeine | 15mg/mL | Prednisone | 0.22mg/mL |
| Chlorophyll | 0.11mg/mL | Protein (albumin) (H) | 10mg/mL |
| Chlorophyll | 0.04mg/mL | Protein (albumin) (M) | 5mg/mL |
| Chlorophyll | 0.01mg/mL | Protein (albumin) (L) | 0.6mg/mL |
| Ciprofloxacin | 0.22mg/mL | Pyridium | 1mg/mL |
| Decongestant | 0.22mg/mL | Rifampicin | 0.09mg/mL |
| Corticosterone (Corticosteroids) | 0.015mg/mL | Spinach | 1% |
| Erythromycin | 0.067mg/mL | Tobacco purified | 0.4mg/mL |
| Glucose (H) | 20mg/mL | Triglycerides | 4mg/mL |
| Glucose (M) | 10mg/mL | Urea | 20mg/mL |
| Glucose (L) | 3mg/mL | Vaginal contraceptive gel | 5% |
| Hemoglobin | 5mg/mL | Vancomycin | 0.1mg/mL |
| Human albumin | 35mg/mL | Water-based personal lubricant | 5% |
| Human red blood cells 10% Washed pooled cells | 10% | White blood cells | 10% |
| Ibuprofen | 0.1mg/mL | Whole blood | 10% |
| Itraconazole | 0.22mg/mL | Whole blood | 15% |
No interference was observed during the study.
## Interfering substances: CSF
ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test was tested with interfering agents at different concentrations. Substances were tested with artificial CSF and artificial CSF spiked with S. pneumoniae antigen at 2-3x LoD. Table 4 list the interfering substances tested.
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Table 4. Interfering substances evaluated in CSF
| Agent | Concentration | Agent | Concentration |
| --- | --- | --- | --- |
| Glucose (H) | 1mg/mL | Glucose (H) | 1mg/mL |
| Glucose (M) | 0.5mg/mL | Glucose (M) | 0.5mg/mL |
| Glucose (L) | 0.1mg/mL | Glucose (L) | 0.1mg/mL |
| Red blood cells (H) | 15% | Red blood cells (H) | 15% |
| Red blood cells(M) | 10% | Red blood cells(M) | 10% |
| Red blood cells (L) | 5% | Red blood cells (L) | 5% |
| Protein (H) | 60mg/mL | Protein (H) | 60mg/mL |
| Protein (M) | 30mg/mL | Protein (M) | 30mg/mL |
| Protein (L) | 10mg/mL | Protein (L) | 10mg/mL |
| White blood cells | 10.6x10^6/mL | White blood cells | 10.6x10^6/mL |
| White blood cells | 5.3x10^6/mL | White blood cells | 5.3x10^6/mL |
| White blood cells | 2.7x10^6/mL | White blood cells | 2.7x10^6/mL |
| White blood cells | 1.8x10^6/mL | White blood cells | 1.8x10^6/mL |
| White blood cells | 0.9x10^6/mL | White blood cells | 0.9x10^6/mL |
| Bilirubin | 15% | Bilirubin | 15% |
| Bilirubin | 10% | Bilirubin | 10% |
| Bilirubin | 5% | Bilirubin | 5% |
| Plasma | 15% | Plasma | 15% |
| Plasma | 10% | Plasma | 10% |
| Plasma | 5% | Plasma | 5% |
No interference was observed during the study.
4. Assay Reportable Range:
Not Applicable
5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):
Sample Stability study
Specimen stability was evaluated for both fresh and preserved samples. Samples were preserved in either Boric acid or piperazine-N,N'-bis(2-ethanesulfonic acid) (PIPES). For the analysis, a panel with a total of 105 samples were prepared as listed in Table 5 below. Low positive samples represented 1 - 2x LoD and Moderate positives represented 4x LoD.
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Table 5. Panel used for Sample Stability Testing
| Panel | Test times | Condition |
| --- | --- | --- |
| Neg urine (N=5) | T0, 1, 2 and 7 days | 2-8 °C and ~30 °C |
| Neg urine with Boric acid (N=5) | | |
| Neg urine with PIPES (N=5) | | |
| | | |
| S. pneumoniae low pos urine (N=10) | | |
| S. pneumoniae low pos urine with Boric acid (N=10) | | |
| S. pneumoniae low pos urine with PIPES (N=10) | | |
| | | |
| S. pneumoniae Mod pos urine (N=5) | | |
| S. pneumoniae Mod pos urine with Boric acid (N=5) | | |
| S. pneumoniae Mod pos urine with PIPES (N=5) | | |
| | | |
| L. pneumophila low pos urine (N=10) | | |
| L. pneumophila low pos urine with Boric acid (N=10) | | |
| L. pneumophila low pos urine with PIPES (N=10) | | |
| | | |
| L. pneumophila Mod pos urine (N=5) | | |
| L. pneumophila Mod pos urine with Boric acid(N=5) | | |
| L. pneumophila Mod pos urine with PIPES (N=5) | | |
Fresh samples were tested at time 0 then stored at refrigeration (2°C - 8°C) and room temperatures (~30°C). Subsequent testing was performed at 1, 2 and 7 days. Testing was conducted following the instructions in the proposed package insert. The results for antigen stability indicated that use of preservative supports stable test performance over the course of 7 days for either refrigeration or room temperature storage. Fresh urine samples remained positive only 83% of the time when tested at 48 hours. Urine samples without preservative must be tested within 24 hours of collection.
## Frozen sample stability
Stability of frozen urine samples compared to fresh samples was established using panels prepared with S. pneumoniae/L. pneumophila antigen at Low (1-2x LoD) and moderate (3-4x LoD) antigen concentrations, S. pneumoniae/L. pneumophila whole organism at low (1-2x LoD) concentration, and true negative samples. Urine specimens were prepared with and without preserved. Samples were stored at -20°C for 7 days and were tested at 0, 1, 2, and 7 days. The results showed that all positive samples remained positive and negative samples remained negative throughout the study.
## Freeze/Thaw study
A study was conducted to determine stability after 5 freeze/thaw cycles using a 110 urine specimen panel described in Table 6. below. Samples were not tested using a preservative. The results showed that up to 5 freeze/thaw cycles did not significantly impact the performance of the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test. The package insert will indicate that if samples are not tested fresh, frozen urine samples may be tested after no more than 5 freeze/thaw cycles.
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Table 6. Freeze/Thaw Panel
| Panel | Replicates |
| --- | --- |
| S. pneumoniae Strong pos (5x LoD) | 10 |
| L. pneumophila Strong pos (5x LoD) | 10 |
| S. pneumoniae Weak pos (1-2 x LoD) | 30 |
| L. pneumophila. Weak pos (1-2 x LoD) | 30 |
| S. pneumoniae High neg (<1 x LoD) | 10 |
| L. pneumophila High neg (<1 x LoD) | 10 |
| True Negative | 10 |
# 6. Detection Limit:
The limit of detection (LoD) for the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test was determined by spiking purified antigen or whole organisms into unpreserved urine and preserved urine (boric acid or PIPES). The concentration of purified antigen was reported in pg/mL for S. pneumoniae and $\mu \mathrm{g} / \mathrm{mL}$ for L. pneumophila, and for whole organisms as CFU/mL. LoD was defined as the concertation the yields at least a positive result in $95\%$ of performed tests. Table 7 lists the LoD values for antigen and whole organism in urine for S. pneumoniae and L. pneumophila. The use of preservatives (boric acid or PIPES) did not significantly impact the LoD.
Table 7. LoD for S. pneumoniae and L. pneumophila
| Test material | LoD |
| --- | --- |
| S. pneumoniae antigen | 62.5 pg/mL |
| L. pneumophila SG 1 (Philadelphia) antigen | 0.025 μg/mL |
| L. pneumophila SG 1 (Bellingham) antigen | 0.5 μg/mL |
| S. pneumoniae (serotype 1) | 10^5 CFU/mL |
| L. pneumophila SG1 (Philadelphia) | 10^4 CFU/mL |
| L. pneumophila SG 1 (Bellingham) | 10^5 CFU/mL |
The LoD in CSF for the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test was determined by spiking whole organisms into clinically negative human CSF. The concentration of S. pneumoniae using whole organisms is reported in CFU/mL. LoD was defined as the concertation the yields at least a positive result in $95\%$ of performed tests in CSF. The study data show that the LoD for S. pneumoniae in CSF is $10^{3}$ CFU/mL. The LoD for L. pneumophila was not evaluated.
# Inclusivity
The reactivity of three additional S. pneumoniae serotypes, 3, 5 and 37, was evaluated. The samples were prepared by spiking clinically negative urine with antigen or whole organism. Testing was conducted around the previously established LoD in replicates of 6 to determine the lowest concentration that provided a positive result $100\%$ of the time.
The reactivity of four Pontiac and three non-Pontiac strains of $L$ pneumophila was evaluated. Samples were prepared by spiking clinically negative urine with antigen or whole organism. Testing was conducted around the previously established the LoD in replicates of 20 or 6 to determine the lowest concentration that provided a positive result $100\%$ of the time. The reactivity of three additional $L$ pneumophila subgroups 3, 6,8,10 and 12, was evaluated. The
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samples were prepared by spiking clinically negative urine with antigen or whole organism. Testing was conducted well above the established LoD in replicates of 6 to determine the lowest concentration that provided a positive result 100% of the time. Table 8 below summarizes the Inclusivity results for S. pneumoniae and L. pneumophila
Table 8. Summary of Inclusivity Testing
| Streptococcus pneumoniae in urine | | | | |
| --- | --- | --- | --- | --- |
| Subgroup | | Antigen Concentration (μg/mL) | Whole Organism Concentration (CFU/mL) | |
| type 1 | | ND* | 10^{4} | |
| type 3 | | 0.001 | 10^{4} | |
| type 5 | | 0.010 | 10^{5} | |
| type 37 | | 0.0001 | ND* | |
| Legionella pneumophila in urine | | | | |
| Subgroup | Pontiac/Non-Pontiac | Species | Concentration (μg/mL) | Concentration (CFU/mL) |
| SG1 | Pontiac | Knoxville | 0.100 | 10^{5} |
| SG1 | Pontiac | Allentown/France | 0.005 | ND* |
| SG1 | Pontiac | Benidorm | ND* | 10^{4} |
| SG1 | Pontiac | Philadelphia | 0.010 | 10^{4} |
| SG1 | Non-Pontiac | OLDA/Oxford | 0.001 | ND* |
| SG1 | Non-Pontiac | Camperdown | 0.315 | ND* |
| SG1 | Non-Pontiac | Heysham | 1.250 | ND* |
| SG3 | | | 250 | ND* |
| SG6 | | | 250 | ND* |
| SG8 | | | 250 | ND* |
| SG10 | | | 250 | ND* |
| SG12 | | | 7.8 | ND* |
*ND = Not tested
7. Assay Cut-Off:
Not Applicable.
B Comparison Studies:
1. Method Comparison with Predicate Device:
See Clinical Studies section below.
2. Matrix Comparison:
Not Applicable.
C Clinical Studies:
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# 1. Clinical Sensitivity:
# Clinical Agreement for Prospective Urine Samples
Prospectively collected patient urine samples $(n = 306)$ from two different sites (Spain and Denmark) were tested with both the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test and two FDA cleared lateral flow urine antigen tests (one per antigen) as comparator assays. Fresh urine samples were from patients at risk of having community acquired pneumonia. The results from each site were pooled and analyzed by analyte. Of the 306 patients, 179 were from Spain and 127 from Denmark. The patient population consisted of 141 females and 165 males, ranging in age from 18-95 years old. The results of this prospective method comparison test for S. pneumoniae antigen found in urine are noted in Table 9. Boiling of samples for discrepant resolution did not change any of the results or performance. Table 10 notes the results of this prospective method comparison test for L. pneumophila antigen found in urine.
Table 9. S. pneumoniae Prospective Study
| ImmuView | Comparator Positive | Comparator Negative | Total |
| --- | --- | --- | --- |
| Positive | 72 | 6 | 78 |
| Negative | 3 | 225 | 228 |
| Positive percent agreement | 96.0% (72/75) | 95% CI (88.9%-98.6%) | |
| Negative percent agreement | 97.4% (225/231) | 95% CI (94.5-98.8%) | |
Table 10. L. pneumophila Serogroup 1 Prospective Study
| ImmuView | Comparator Positive | Comparator Negative | Total |
| --- | --- | --- | --- |
| Positive | 3 | 0 | 3 |
| Negative | 0 | 303 | 303 |
| Positive percent agreement | 100.0% (3/3) | 95% CI (43.9%-92.9%) | |
| Negative percent agreement | 100.0% (303/303) | 95% CI (98.8%-100%) | |
# Clinical Sensitivity/Specificity for Retrospective Urine Samples
To further characterize test performance, 100 frozen urine samples from patients previously determined to be S. pneumoniae blood culture positive were tested. In total there were 48 samples from Sweden and 52 samples from Denmark. These results are described in Table 11. Discrepant results were boiled, and the results are footnoted below.
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Table 11. S. pneumoniae Retrospective Sample Testing (urine)
| ImmuView | Blood Culture Positive | Blood Culture Negative | Total |
| --- | --- | --- | --- |
| Positive | 78^{a} | 4 | 82 |
| Negative | 22 | 217^{b} | 239 |
| Sensitivity | 78.0% (78/100) | 95% CI (68.9%-85%) | |
| Specificity | 98.1% (217/221) | 95% CI (95.4%-99.3%) | |
a After boiling three samples turned positive (81/100)
b After boiling one false positive turned negative (218/221)
To further characterize test performance, 98 stored frozen urine samples from patients previously determined to be Legionella respiratory culture positive. A total of 55 urine samples were from Europe and 43 urine samples were from patients within the United States. These results are described in Table 12.
Table 12. L. pneumophila Serogroup 1 Retrospective Sample Testing (urine)
| ImmuView | Sputum Culture Positive | Sputum Culture Negative | Total |
| --- | --- | --- | --- |
| Positive | 86^{c} | 1 | 87 |
| Negative | 12 | 239^{d} | 251 |
| sensitivity | 87.8% (86/98) | 95% CI (79.8%-92.9%) | |
| specificity | 99.6% (239/240) | 95% CI (97.7%-99.9%) | |
c After boiling two samples turned negative (84/98)
d After boiling one sample tuned negative (240/240)
The clinical specificity of the test was established by testing known negative (culture confirmed negative) urine samples collected from 3 sites, one in the U.S. and two in Europe.
## Clinical sensitivity and specificity - CSF
The sensitivity of the S. pneumoniae test line was obtained by testing leftover (retrospective) CSF specimens from US and European patients suspected of meningitis. Testing was conducted for contrived specimens were of human CSF spiked with antigen and negative CSF.
## Retrospective specimens.
Nine CSF specimens known positive for S. pneumoniae were tested at two U.S. labs with 113 samples previously determined to be negative. Samples were blinded and the testing was performed by three operators on different days. Five CSF specimens known positive for S. pneumoniae were tested at one European laboratory with 56 samples previously determined to be negative. Samples were blinded and tested by one operator on multiple days. Results are in table 13 below.
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Table 13. Retrospective CSF test results
| ImmuView | S. pneumoniae Culture positive (CSF) | S. pneumoniae Culture negative (CSF) | Total |
| --- | --- | --- | --- |
| S. pneumoniae Positive | 13 | 7 | 20 |
| S. pneumoniae Negative | 1 | 162 | 163 |
| Sensitivity | 92.9% (13/14) | 95% CI (68.5%-98.7%) | |
| Specificity | 95.9% (162/169) | 95% CI (91.7%-98.0%) | |
The sensitivity of device $L$ pneumophila test line was not evaluated in this study as Legionella is an exceptionally rare etiology for meningitis, and CSF is not included as a matrix for meningitis testing in the device Intended Use.
# Contrived CSF testing
Additional human CSF testing was conducted using whole organisms spiked into human CSF. Testing was conducted with the ImmuView test and the comparator for the following contrived samples: 25 samples with S. pneumoniae whole organism near the LoD, 25 samples with S. pneumoniae whole organism across a clinically relevant range, and 10 negative CSF samples. The positive percent agreement and negatieve percent agreement for the both the ImmuView test and the comparator test was $100\%$ , 50/50 for PPA and 10/10 for NPA. Table 14 summarizes these results.
Table 14 Contrived CSF results (S. pneumoniae whole organism spiked into human CSF)
| ImmuView | S. pneumoniae Contrived Positive | S. pneumoniae Contrived Negative | Total |
| --- | --- | --- | --- |
| ImmuView S. pneumoniae Positive | 50 | 0 | 50 |
| ImmuView S. pneumoniae Negative | 0 | 10 | 10 |
| Positive percent agreement | 100% | 95% CI (92.9%-100%) | |
| Negative percent agreement | 100% | 95% CI (72.2%-100%) | |
2. Clinical Specificity:
See section C.1. above.
3. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
See section C.1. above.
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D Clinical Cut-Off:
Not applicable.
E Expected Values/Reference Range:
Not applicable.
VIII Proposed Labeling:
The labeling supports the finding of substantial equivalence for this device.
IX Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
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