Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device is described as a rapid lateral flow immunochromatographic assay, which is a traditional diagnostic technology that does not typically incorporate AI/ML. There are no mentions of AI, ML, deep learning, or image processing in the summary. The performance studies focus on traditional metrics like sensitivity and specificity, not metrics typically associated with AI/ML model evaluation.

Therapeutic

No
The device is described as a diagnostic test for detecting antigens, which aids in diagnosis rather than providing therapy or treatment.

Diagnostic

Yes

Explanation: The "Intended Use / Indications for Use" section explicitly states that the device is "intended to aid in diagnosis" of specific infections. The "Device Description" also mentions its effectiveness in "presumptive diagnosis".

SaMD (Software as a Medical Device)

No

The device is described as a "rapid, lateral flow test, also known as a lateral flow immunochromatographic assay," which is a physical test strip and not a software-only device.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device is an "in vitro, rapid, lateral flow test" intended for the "qualitative detection of Streptococus pneumoniae and Legionella pneumophila antigens in urine specimens" and "S. pneumoniae antigen in cerebrospinal fluid (CSF)". This clearly indicates that the device is used to examine specimens taken from the human body to provide information for the diagnosis of disease.
Device Description: The "Device Description" further reinforces this by stating it's a "rapid lateral flow test for qualitative detection of S. pneumoniae in human urine and CSF samples and L. pneumophila... antigens in human urine samples." It also mentions its use in the "presumptive diagnosis" of pneumococcal pneumonia and Legionnaires' disease.
Performance Studies: The inclusion of detailed performance studies (Clinical Sensitivity and Specificity, Positive and Negative Percent Agreement) using human urine and CSF samples is characteristic of an IVD device undergoing regulatory review.
Predicate Device(s): The listing of predicate devices, which are also IVDs (BinaxNOW® Streptococcus pneumoniae and BinaxNOW® Legionella), further confirms the device's classification as an IVD.

All these points align with the definition and characteristics of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / 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.

Product codes (comma separated list FDA assigned to the subject device)

MJH, GTZ

Device Description

ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test is a rapid lateral flow test for 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 effective in presumptive diagnosis of pneumococcal pneumonia caused by S. pneumoniae or Legionella pneumonia (Legionnaires' disease) caused by L. pneumophila, in conjunction with culture and other methods. Correct and early treatment is vital for the prognosis of both diseases and therefore quick methods to confirm both diseases in the initial phase are very important in order to initiate the proper antibiotic treatment as soon as possible.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Urine, Cerebrospinal Fluid (CSF)

Indicated Patient Age Range

Not Found

Intended User / Care Setting

This device is for in vitro diagnostic use only. It is for prescription use only and to be used only by clinical laboratory professionals.

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Clinical Sensitivity and Specificity for Urine Samples (Retrospective study)

S. pneumoniae: 100 frozen urine samples from patients originally determined to be infected with S. pneumoniae. All 100 samples were from Europe (48 from Sweden, 52 from Denmark) and from blood culture positive patients.
L. pneumophila: 98 stored frozen urine samples from patients with a culture confirmed Legionella infection. 55 samples from Europe, 43 from the U.S. (previously positive in a urinary antigen test).
Specificity (Urine): Known negative (culture confirmed negative) urine samples collected from 3 sites (one in the U.S., two in Europe).
Annotation protocol: Culture confirmed.

Positive and Negative Percent Agreement for urine samples (Prospective study)

Sample size: 306 prospective collected urine samples.
Data source: Two different sites (Spain and Denmark). Fresh* urine samples were from patients (all comers) at risk of having community acquired pneumonia.
Annotation protocol: Results were compared with other lateral flow urine antigen tests (Comparator) implicitly serving as the reference.
*Of the 306 samples, a total of 92 had to be frozen before testing could be performed.

Clinical sensitivity and specificity - CSF

Sensitivity: Leftover CSF specimens from patients suspected of meningitis, as well as spiked CSF and negative CSF.
- U.S.A Laboratory testing: 9 samples known positive for S. pneumoniae meningitis. 113 negative human CSF samples. Blinded, testing performed by three operators on different days to prevent test bias.
- European Laboratory testing: 5 samples known to be positive for S. pneumoniae. 56 negative CSF samples. Blinded, testing performed by one operator on different days to prevent test bias.
Spiked CSF testing: Additional human CSF samples were spiked at the LOD with S. pneumoniae (N=50) and an additional unspiked negative CSF samples (N=10).

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Clinical Sensitivity and Specificity for Urine Samples (Retrospective study)

Study Type: Retrospective
Sample Size: S. pneumoniae (100 culture-positive, 221 culture-negative); L. pneumophila (98 culture-positive, 240 culture-negative).
Key Results:
- S. pneumoniae:
  - Sensitivity: 78% (78/100) (95%CI 69.0-85.0%)
  - Specificity: 98.1% (217/221) (95%CI 95.4-99.3%)
  - Comparison to Comparator (Blood culture only): ImmuView 78% (78/100) CI 67-85% vs. Comparator 80% (76/95) CI 71-87%.
  - After boiling: ImmuView S. pneumoniae sensitivity (Europe) increased to 81% (81/100).
  - Specificity (Europe) increased to 98.6% (218/221) after boiling.
- L. pneumophila:
  - Sensitivity: 87.8% (86/98) (95%CI 79.8-92.9%)
  - Specificity: 99.6% (239/240) (95%CI 97.4-100%)
  - Comparison to Comparator:
    - L. pneumophila Sg 1 (U.S.): ImmuView 97.7% (42/43) CI 88-100% vs. Comparator 100% (43/43) CI 92-100%.
    - L. pneumophila Sg 1 (Europe): ImmuView 80.0% (44/55) CI 68-88% vs. Comparator 66.7% (36/54) CI 53-78%.
  - After boiling: L. pneumophila sensitivity (U.S.) increased to 100% (43/43) (95%Cl 91.8-100%) in the ImmuView Test. L. pneumophila sensitivity (Europe) changed to 74.6% (41/55). Specificity (Europe) increased to 100% (221/221) after boiling.

Positive and Negative Percent Agreement for urine samples (Prospective study)

Study Type: Prospective agreement study
Sample Size: 306
Key Results:
- S. pneumoniae:
  - 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%) (Note: text states 226/232 or 97.4%)
- L. pneumophila SG1:
  - Positive percent agreement: 100.0% (3/3) (95% CI 43.9%-100%)
  - Negative percent agreement: 100.0% (303/303) (95% CI 98.8%-100%) (Note: text states 304/304 or 100%)
- After boiling, positive and negative agreement remained the same.

Analytical Studies - Urine

Specificity (Cross-Reactivity): Tested urines spiked with 143 whole cell bacteria and different inactivated viruses. All bacterial isolates were negative except 1 of 3 isolates of E. cloaced which was positive for L. pneumophila. No cross-reactions with 19 urinary tract infection samples.
Sensitivity (Limit of detection (LOD)):
- S. pneumoniae CWPS antigen (native): 62.5 pg/mL
- LPS specific for L. pneumophila SG1 (Philadelphia): 25 ng/mL (0.025 µg/mL)
- Whole cell S. pneumoniae bacteria: 10^5 CFU/mL
- L. pneumophila SG1 (Philadelphia): 10^4 CFU/mL
- L. pneumophila SG 1 (Bellingham): 10^5 CFU/mL
- L. pneumophila SG 1 (Bellingham) antigen: 0.5 µg/mL
Strain Reactivity:
- S. pneumoniae serotypes: positive for type 1, 3, 5, 37.
- L. pneumophila: positive for SG1 (Philadelphia, Knoxville, OLDA/Oxford, Allentown/France, Benidorm-Strain Lens) and serogroups 3, 6, 8, 10, 12.
Interfering Substances: Tested with 47 interfering agents. High concentration of plasma may result in gray test lines. Basic (pH≥9) conditions may give false positive S. pneumoniae lines. Water-based personal lubricant might result in false positive or gray L. pneumophila lines (dose-related).

Clinical sensitivity and specificity - CSF

Study Type: Clinical sensitivity and specificity
Sample Size: 14 S.pn. Culture positive, 169 S.pn. Culture negative.
Key Results:
- Sensitivity: 92.9% (13/14) (95% CI 68.5%-98.7%)
- Specificity: 96.0% (162/169) (95% CI 91.7%-98.0%)
Spiked CSF testing:
- Sample Size: 60 real human CSF samples (50 spiked with S. pneumoniae, 10 unspiked negative).
- Key Results: ImmuView and comparator both showed 100% positive percent agreement (50/50, 95% CI 92.9%-100%) and 100% negative percent agreement (10/10, 95% CI 72.2%-100%).

Analytical Studies - CSF

Specificity (Cross-Reactivity): Tested with 24 potential cross-reacting agents. No cross-reactions detected for S. pneumoniae or L. pneumophila test lines.
Sensitivity (Limit of detection (LOD)) in CSF: S. pneumoniae: 10^3 CFU/mL.
Interference agents: Tested with 47 interfering agents. Red blood cells may give false positive shadows on the S. pneumoniae line. Other agents did not interfere.

Reproducibility study

Study Type: Reproducibility
Sample Size: 1,072 test results (from an initial 1,080 tests).
Data Source: 10 members of real positive S. pneumoniae or L. pneumophila urine samples, negative urine samples, artificial CSF positive spiked with S. pneumoniae isolates, negative artificial CSF samples. ImmuView Positive Control and Negative Control.
Key Results:
- Overall Reproducibility: 99.6% (1,068 correct results out of 1,072).
- Ranges from 98.9% to 100.0% for individual categories (e.g., S. pneumoniae moderate positive urine 100%, L. pneumophila low positive urine 1B 98.9%, ImmuView Pos Control 98.9%, ImmuView Neg Control 100%).
- Performed for 5 days with a different kit lot at each site (two in U.S., one in Europe). 3 different lots tested. 6 operators involved.
- No statistical differences in reproducibility by lot, by time or by operator.
- 8 test results (0.7%) were invalid and excluded.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Clinical Sensitivity and Specificity for Urine Samples (Retrospective study)

S. pneumoniae Sensitivity: 78% (95%CI 69.0-85.0%)
S. pneumoniae Specificity: 98.1% (95%CI 95.4-99.3%)
L. pneumophila Sensitivity: 87.8% (95%CI 79.8-92.9%)
L. pneumophila Specificity: 99.6% (95%CI 97.4-100%)

Positive and Negative Percent Agreement for urine samples (Prospective study)

S. pneumoniae Positive percent agreement: 96.0% (95% CI 88.9%-98.6%)
S. pneumoniae Negative percent agreement: 97.4% (95% CI (94.5%-98.8%)
L. pneumophila SG1 Positive percent agreement: 100.0% (95% CI 43.9%-100%)
L. pneumophila SG1 Negative percent agreement: 100.0% (95% CI 98.8%-100%)

Clinical sensitivity and specificity - CSF

S. pneumoniae sensitivity (CSF): 92.9% (95% CI 68.5%-98.7%)
S. pneumoniae specificity (CSF): 96.0% (95% CI 91.7%-98.0%)
Spiked CSF testing Positive percent agreement: 100% (95% CI 92.9%-100%)
Spiked CSF testing Negative percent agreement: 100% (95% CI 72.2%-100%)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

K012521, K982238

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

K991726, K070522

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

Regulation Number and Section

§ 866.3300

Haemophilus spp. serological reagents.(a)
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 identifyHaemophilus 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 genusHaemophilus and provides epidemiological information on diseases cause by these microorganisms. Diseases most often caused byHaemophilus spp. include pneumonia, pharyngitis, sinusitis, vaginitis, chancroid venereal disease, and a contagious form of conjunctivitis (inflammation of eyelid membranes).(b)
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.

Summary

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

March 5, 2020

SSI Diagnostica A/S % Christopher Bentsen Regulatory and Clinicals Consultant Bentsen Regulatory and Clinicals Consulting LLC 25803 NE 9th Street Redmond, Washington 98074

Re: K191184

Trade/Device Name: ImmuView S pneumoniae and L pneumophila Urinary Antigen Test Regulation Number: 21 CFR 866.3300 Regulation Name: Haemophilus Spp. Serological Reagents Regulatory Class: Class II Product Code: MJH, GTZ Dated: June 3, 2019 Received: June 10, 2019

Dear Christopher Bentsen:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal

1

statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Kristian Roth, Ph.D. Chief Bacterial Multiplex and Medical Counter Measures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K191184

Device Name

ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test

Indications for Use (Describe)

Intended 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.

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

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510(k) #	K191184	510(k) SUMMARY	Date of Preparation: March 4, 2020
Submitter:	Christopher Bentsen, M.S., RAC, FRAPS; Bentsen Regulatory and Clinicals Consulting LLC.
Submitters Address:	Gig Harbor, Washington 98332
Submitters Number:	(206) 910-1974
Sponsor:	SSI Diagnostica A/S (SSID)
Contact:	Dr. Pernille Landsbo Elverdal, VP R&D
Contact Number:	0045 4111 2731
Device Name:	ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test
Common Name:	Urinary Antigen Test, Streptococcus pneumoniae and Legionella pneumophila serogroup 1
Classification:	Streptococcus spp and Legionella spp serological reagents
21 CFR 866.3740 and 866.3300
Product Codes:	GTZ and MJH
Predicate Devices:	BinaxNOW® Streptococcus pneumoniae (K012521) and BinaxNOW® Legionella (K982238)

Panel:

Microbiology

Intended 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 Streptocccus 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 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.

Indication(s) for use:

Same as Intended Use

Special conditions for use statement(s):

This device is for in vitro diagnostic use only. It is for prescription use only and to be used only by clinical laboratory professionals.

Special instrument requirements:

N/A

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Device Description:

ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test is a rapid lateral flow test for 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 effective in presumptive diagnosis of pneumococcal pneumonia caused by S. pneumoniae or Legionella pneumonia (Legionnaires' disease) caused by L. pneumophila, in conjunction with culture and other methods. Correct and early treatment is vital for the prognosis of both diseases and therefore quick methods to confirm both diseases in the initial phase are very important in order to initiate the proper antibiotic treatment as soon as possible.

Substantial Equivalence Information:

1. Predicate device names(s): BinaxNOW" Streptococcus pneumoniae Card and BinaxNOW" Legionella pneumophila Card
1. Predicate 510(k) number(s): K991726 and K012521 and K982238 and K070522

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Table 1 Comparison with predicate:

| Description | ImmuView S. pneumoniae and
L. pneumophila Urinary
Antigen Test | BinaxNOW® Streptococcus
pneumoniae and BinaxNOW®
Legionella Antigen Cards | |
|------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------|
| Test Format | Rapid
immunochromatographic
lateral flow test | Rapid immunochromatographic
card tests | |
| Quantitative/Quantitative | Qualitative | Qualitative | |
| Test Antigen | Streptococcus pneumoniae
and Legionella pneumophila
serogroup 1 | Streptococcus pneumoniae or
Legionella pneumophila
serogroup 1 | |
| Specimen Types | Human CSF and human urine | Human CSF and human urine | |
| Reagents/Components | Test Strips
Test Tubes
Sample Running Buffer
Negative Control
Positive Control
Plastic transfer pipettes
Tweezer
Cardboard test tube holder | Test Cards
Reagent A
Positive Control Swab
Negative Control Swab
Swabs | |
| Antibody Sources | | | |
| Test Card | Rabbit Polyclonal antibodies | Rabbit Polyclonal antibodies | |
| Conjugate | Rabbit Polyclonal antibodies | Rabbit Polyclonal antibodies | |
| Sample Preparation | | | |
| Unpreserved and
preserved urine | Add 3 (120 ul) drops of urine
or add 10 ul of CSF to a test
tube
Add 2 (90 ul) drops of sample
running diluent
Mix well
Add test strip with arrow
down | 1. Dip swab into the urine sample
and then insert swab into the
bottom hole of the Test Card.
2. Add 2 drops of Reagent A to
the bottom hole for Legionella
Card and 3 drops for S.
pneumoniae Card. | |
| Testing Time | Approximately 15 minutes | Approximately 15 minutes | |
| Equipment | | | |
| General Laboratory
Equipment | Urine collection container
Timer
Vortex or mix by swirling
Disposable gloves | Urine collection container
Timer
Swab pack
Disposable gloves | |
| Reading Method | Visual | Visual | |
| Results Interpretation | | | |
| Visual Read | Negative: A single purple/gray
Control line in the top of the
strip.
Positive: For both S. pn. and L.
pn. S-1 will show a pink/red
line and a blue line.
For S. pn. will show a pink/red
line.
For L. pn. S-1 will show a blue
line | Negative: Single pink to purple
colored Control line visible in top
half of the window
Positive: Two pink to purple lines | |
| | Invalid: No line at the Control line position and if there is a
dot instead of a test line.
Also, if gray lines appear.
Follow IFU instructions | Invalid: No line at the Control line position or no lines at the Control and Sample line positions. | Test |

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Principle:

ImmuView® S. pneumoniae and L. pneumophila Urinary Antigen Test is a rapid lateral flow test for detection of S. pneumoniae and L. pneumophila using the same test.

Clinical Sensitivity and Specificity for Urine Samples (Retrospective study)

To determine the sensitivity of the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test, 100 frozen urine samples from patients originally determined to be infected with S. pneumoniae were tested. All 100 urine samples came from Europe, and all were from blood culture positive patients; Fortyeight (48) samples were from Sweden3 and fifty-two (52) samples were from Denmark.

To determine the sensitivity of the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test, 98 stored frozen urine samples from patients with a culture confirmed Legionella infection were tested. A total of 55 urine samples came from Europe. The remaining 43 urine samples came from the United States (U.S.), and these were also determined to be previously positive in a urinary antigen test.

The clinical specificity of the ImmuView S. pneumoniae and L. pneumophila test lines was obtained by testing known negative (culture confirmed negative) urine samples collected from 3 sites, one in the U.S. and two in Europe.

S. pneumoniae culture verified vs. ImmuView
	Culture positive	Culture negative
ImmuView pos	78	4
ImmuView neg	22	217
Total	100	221
ImmuView Sensitivity	78%	95%CI (69.0-85.0%)
ImmuView Specificity	98.1%	95%CI (95.4-99.3%)
L. pneumophila culture verified vs. ImmuView
	Culture positive	Culture negative
ImmuView pos	86	1
ImmuView neg	12	239
Total	98	240
ImmuView Sensitivity	87.8%	95%CI (79.8-92.9%)

Table 1

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Table 3

	ImmuView	Comparator
Sensitivity (Urine) Based on culture vs comparator
S. pneumoniae
(Blood culture only)	78% (78/100)
(CI 67-85%)	80% (76/95a)
(CI 71-87%)
L. pneumophila Sg 1
(U.S.)	97.7% (42/43)
(CI 88-100%)	100% (43/43)
(CI 92-100%)
L. pneumophila Sg 1
(Europe)	80.0% (44/55)
(CI 68-88%)	66.7% (36/54b)
(CI 53-78%)
Specificity (Urine) Based on culture vs comparator
S. pneumoniae
(Europe)	98.2% (217/221c)
(CI 95-99%)	97.8% (218/223)
(CI 95-99%)
L. pneumophila
(U.S.)	100% (19/19)
(CI 83-100%)	100% (19/19)
(CI 83-100%)
L. pneumophila
(Europe)	99.5% (220/221d)
(CI 97-100%)	99.6% (223/224)
(CI 98-100%)

ª 5 samples were QNS for testing, º 1 sample was QNS for testing, º 3 samples were QNS for testing, º 3 samples were QNS for testing

S. pneumoniae sensitivity (Europe) increased to 81/100 or 81% for ImmuView S. pneumoniae and L. pneumophila urinary antigen test compared with comparator that after boiling had 76/95 or 80%. L. pneumophila sensitivity (Europe) changed to 41/55 or 74.6% for ImmuView and remained 36/54 or 66.7% for the comparator. The specificity (Europe) increased to 98.6% (218/221) and 100% (221/221) for S. pneumoniae and L. pneumophila respectively after boiling when using ImmuView. The comparator did not change after boiling. L. pneumophila sensitivity (U.S.) increased to 43/43 or 100% (95%Cl 91.8-100%) in the ImmuView Test for L. pneumophila after boiling2-2. L. pneumophila specificity (U.S.) did not change after boiling for either test.

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Positive and Negative Percent Agreement for urine samples

(Prospective study)

In a prospective study three-hundred-six (306) prospective collected urine samples from two different sites (Spain and Denmark) were tested with both the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test and the Comparator tests. Fresh* urine samples were from patients (all comers) at risk of having community acquired pneumonia. The results were compared with other lateral flow urine antigen tests (Comparator).

| able
T

4
----------------

Prospective samples positive agreement S. pneumoniae
ImmuView	Comparator positive	Comparator negative	Total
Positive	72	6	78
Negative	3	225	228
Total	75	231	306
Positive percent agreement	96.0%	95% CI (88.9%-98.6%)
Negative percent agreement	97.4%	95% CI (94.5%-98.8%)
Prospective samples positive agreement L. pneumophila SG1
ImmuView	Comparator positive	Comparator negative	Total
Positive	3	0	3
Negative	0	303	303
Total	3	303	306
Positive percent agreement	100.0%	95% CI (43.9%-100%)
Negative percent agreement	100.0%	95% CI (98.8%-100%)

Of the 306 samples, a total of 92 had to be frozen before testing could be performed.

The positive agreement for S. pneumoniae was 72/75 or 96% (88.9-98.6%). The negative agreement for S. pneumoniae was 226/232 or 97.4% (94.5-98.8). The positive agreement for L. pneumophila was 3/3 or 100% (43.9-100%). Negative agreement for L. pneumophila was 304/304 or 100% (98.8-100%).

After boiling22 the positive and negative agreement for S. pneumoniae and L. pneumophila remained the same.

10

Analytical Studies - Urine

Specificity (Cross-Reactivity)

To determine the analytical specificity of the ImmuView S. pneumoniae and L. pneumophila Urinary Antigen test for cross-reactivity with urines spiked with whole cell bacteria and different inactivated viruses (N=143). The whole cell bacterial panel was tested in a 107 CFU/mL diluted from a stock solution. The Viral panel had a concentration of 105 TCID50/mL. The panel was also tested in negative urine.

Organisms tested for interference
Acinetobacter ssp.(4)	Lactobacillus sp.
Bacillus subtilis	Listeria monocytogenes
Bordetella pertussis	Morganella morganii
Moraxella catarrhalis	Moraxella osloensis
Candida albicans (4)	Mycoplasma genitalium
Citrobacter freundii	Neisseria gonorrhoeae (3)
Cornyebacterium sp.	Neisseria lactamica
Cornyebacterium uralyticum	Neisseria meningitidis
Enterobacter cloacae (3)	Neisseria polysaccharea
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)
Haemophilus Influenzae type a-f and non-caps (11)	Salmonella bredeney
Haemophilus paraInfluenzae	Salmonella Thompson
Adenovirus 2,	Salmonella typhimurium
Chlamydophila pneumoniae (2)	Serratia marcescens
Chlamydia trachomatis	Staphylococcus epidermidis
Cytomegalovirus	Salmonella glostrup
Enterovirus D68	Streptococcus mutans (2)
Herpes Simplex 1,2	Streptococcus parasanguis
Influenzae A (H1N1 and H3N2) virus	Streptococcus sanguinis
Influenzae B Virus	Streptococcus aureus (6)
ParaInfluenzae 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)
Lactobacillus catenaforme	Streptococcus mitis
Lactobacillus rhamnosus

Table 5

All of the above bacterial isolates were negative when using ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test. The only potential cross-reactivity was 1 of 3 isolates of E. cloaced which was positive for L. pneumophila. This was confirmed on re-testing of that one isolate.

A total of 19 Urinary tract infection from patients were tested. Previously culture results had shown that eight (8) of them were infected with Escherichia coli, five (5) with Staphylococcus, aureus, five (5) with Streptococcus agalactiae gr. B and one (1) with Candida albicans. None showed any cross reactions with the ImmuView test.

11

Sensitivity (Limit of detection (LOD))

The limit of detection (LOD) for the ImmuView S. pneumoniae and L. pneumophila urinary antigen test is 62.5 pg/mL for purified S. pneumoniae CWPS antigen (native). For LPS specific for L. pneumophila SG1 (Philadelphia) the LOD is 25 ng/mL. Whole cell S. pneumoniae bacteria can be detected at an LOD at 105 CFU/mL and L. pneumophila SG1 (Philadelphia) has a LOD at 104 CFU/mL. Boiling or urine preservatives did not change these results.

Table 6

Stock solution	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)	105 CFU/mL
L. pneumophila SG1 (Philadelphia)	104 CFU/mL
L. pneumophila SG 1 (Bellingham)	105 CFU/mL

Strain Reactivity

lsolates from different S. pneumoniae serotypes were also positive tested with the ImmuView assay including serotype three (3), five (5), and thirty-seven (37). Different species of L. pneumophila were also found to be positive using the assay. Within serogroup one (1) these includes Philadelphia, Knoxville, OLDA/Oxford, Allentown/France, and Benidorm-Strain Lens. In additional studies have found other Legionella serogroups to be positive such as serogroup 3, 6, 8, 10 and 12.

Streptococcus pneumoniae in urine
Subgroup	Antigen Concentration
(µg/mL)	Whole Organism Concentration
(CFU/mL)
type 1	ND*	104
type 3	0.001	104
type 5	0.010	105
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	105
SG1	Pontiac	Allentown/France	0.005	ND*
SG1	Pontiac	Benidorm	ND	104
SG1	Pontiac	Philadelphia	0.010	104
SG1	Non-Pontiac	OLDA/Oxford	0.001	ND
SG1	Non-Pontiac	Camperdown	0.315	ND
SG1	Non-Pontiac	Heysham	1.250	ND

12

SG3	250	ND
SG6	250	ND
SG8	250	ND
SG10	250	ND
SG12	7.8	ND

*ND=Not done

13

Interfering Substances

ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test were tested with forty-seven (47) interfering agents at different concentrations in urine samples.

Table 8

Agent	Concentration	Agent	Concentration
Acetaminophen	0.1mg/mL	Leucocytes	>250 cells/µL
Acetylsalicylic 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%	10%	White blood cells	10%
Washed pooled cells
Ibuprofen	0.1mg/mL	Whole blood	10%
Itraconazole	0.22mg/mL	Whole blood	15%

High concentration of plasma in urine may result in gray test lines. Additionally, basic (pH≥9) conditions in urine can give false positive S. pneumoniae lines. Water-based personal lubricant might result in false positive or gray L. pneumophila lines, however, this outcome seems dose-related.

14

Clinical sensitivity and specificity - CSF

The sensitivity of the S. pneumoniae test line was obtained by testing leftover CSF specimens from patients suspected of meningitis, as well as spiked CSF and negative CSF table 10 below.

Table 9

ImmuView	S.pn. Culture positive	S.pn. Culture negative
S.pn. Positive	13	7
S.pn. Negative	1	162
Total	14	169
sensitivity	92.9% (13/14)	95% CI (68.5%-98.7%)
specificity	96.0% (162/169)	95% CI (91.7%-98.0%)

U.S.A Laboratory testing

Of the samples tested at the two U.S. labs, 9 were known positive for S. pneumoniae meningitis. Onehundred-thirteen (113) were negative human CSF samples were blinded, and the testing of the ImmuView Test was performed by three operators on different days to prevent test bias.

European Laboratory testing

Of the samples tested within Europe, 5 were known to be positive for S. pneumoniae. Of the total samples, 56 were negative CSF samples were blinded and the testing with the ImmuView Test was performed by one operator on different

days to prevent test bias.

The sensitivity of ImmuView L. pneumophila test line was not validated in this study, Legionella do not usually cause meningitis.

Spiked CSF testing

Additional human CSF samples were spiked at the LOD with S. pneumoniae (N=50) and an additional unspiked negative CSF samples (N=10) were tested with the Immuview test and the comparator test. The sensitivity for the both the ImmuView test and the comparator test was 50/50 (100%) and the additional negative CSF samples used for blinding of the testing were negative 10/10 (100%) in both the ImmuView test and the comparator test.

60 real human CSF samples 50 spiked with S. pneumoniae
ImmuView	Comparator
	S.pn. Positive	S.pn. Negative	Total
S.pn. Positive	50	0	50
S. pn. Negative	0	10	10
Positive percent
agreement	100%	95% CI (92.9%-100%)
Negative percent
agreement	100%	95% CI (72.2%-100%)

15

Analytical Studies - CSF

Specificity (Cross-Reactivity)

ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test were tested with a panel of 24 potential cross-reacting agents. No cross-reactions were detected for the S. pneumoniae or the L. pneumophila test lines.

Table 11

Organisms not affecting test performance in CSF
E. coli (5)	Neisseria meningitidis Gr. B, D and W135 (3)
Haemophilus influenza type a-f and non-caps (7)	Staphylococcus aureus
Listeria monocytogenes	Streptococcus Gr A
Measles	Streptococcus agalactiae (GBS) sg la, Ib, II, III (4)
	Streptococcus mitis

Sensitivity (Limit of detection (LOD)) in CSF

ImmuView S. pneumoniae and L. pneumophila analytical sensitivity was determined by limit of detection. Two different operators performed the dilutions and the testing. The dilutions were made with whole cell bacteria spiked in human CSF.

CSF	LoD
S. pneumoniae	103 CFU/mL

16

Interference agents

ImmuView S. pneumoniae and L. pneumophila Urinary Antigen Test were tested with forty-seven (47) interfering agents at different concentrations in artificial CSF either negative or spiked with either CWPS or S. pneumoniae 107 CFU/mL.

Table 13

Agent in CSF	Concentration	Agent	Concentration
Whole S. pneumoniae (Type 1)		Negative Artificial CSF
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	10.6x106/mL	White blood cells	10.6x106/mL
White blood cells	5.3x106/mL	White blood cells	5.3x106/mL
White blood cells	2.7x106/mL	White blood cells	2.7x106/mL
White blood cells	1.8x106/mL	White blood cells	1.8x106/mL
White blood cells	0.9x106/mL	White blood cells	0.9x106/mL
		Bilirubin
Antigen		Bilirubin
Bilirubin	15%	Bilirubin
Bilirubin	10%	Plasma
Bilirubin	5%	Plasma
Plasma	15%	Plasma
Plasma	10%
Plasma	5%

Red blood cells may give false positive shadows on the S. pneumoniae line due to excessive red color. The other agents in the panel did not interfere with the test.

17

Reproducibility study

The ImmuView S. pneumoniae and L. pneumophila Urinary Antigen test demonstrated excellent overall reproducibility with 1,068 correct results out of 1,072 test results (99.6%), when tested with 10 members of real positive S. pneumoniae or L. pneumophila urine samples and negative urine samples; and artificial CSF positive spiked with S. pneumoniae isolates as well as negative artificial CSF samples. The ImmuView Positive Control and Negative Control were also tested as blinded/masked panel members. The testing was performed for 5 days with a different kit lot at each site, two in the U.S. and one in Europe.

Description	Correct results	Agreement
S. pneumoniae , moderate positive urine	90/90 Positive	100.0%
S. pneumoniae , moderate positive CSF	89/892 Positive	100.0%
S. pneumoniae , low positive spiked in artificial CSF	89/903 Positive	98.9%
S. pneumoniae , low positive urine	90/90 Positive	100.0%
L. pneumophila , moderate positive urine 2A	90/90 Positive	100.0%
L. pneumophila , moderate positive urine 2B	88/894 Positive	98.9%
L. pneumophila , low positive urine 1A	89/895 Positive	100.0%
L. pneumophila , low positive urine 1B	89/906 Positive	98.9%
Negative pooled urine	90/90 Negative	100.0%
Negative artificial CSF	90/90 Negative	100.0%
ImmuView Pos Control	89/907 Positive	98.9%
ImmuView Neg Control	85/858 Negative	100.0%
Summary	1068/1072 Correct	99.6%

Table 14

A total of 3 different lots were tested. Each site, using two operators (A and B) performed a total of 360 reproducibility tests and a grand total of 1,072 reproducibility results out of 1,080 tests in the study using 6 operators. A total of 8 test results (0.7%) were determined to be invalid and were excluded and not re-tested. The panel members were blinded by changing of the panel members and identity daily. The reading and interpretation of the reproducibility panels was performed visually. There were no statistical differences in reproducibility by lot, by time or by operator.

18

The protocol was run five different days, each day each sample had a different code number.
Operator did not see a positive control band, so one sample was invalid as the package insert states that this is necessary before interpreting the result. The sample was not re-tested.
A visual L. pneumophila band was seen.
Operator interpreted band as S. pneumoniae positive instead of L. pneumophila positive. One sample was invalid due to dot (incomplete band) on the strip per the package insert and was not re-tested.
One sample was invalid due to an incomplete band in S. pneumoniae according to the pack insert.
No L. pneumophila band present.
Operator interpreted S. pneumoniae Band result as negative even though band was present.
Five samples excluded due to the presence of dots and incomplete bands. The samples were not retested.

19

References

Rota MC, Fontana S, Montaño-Remacha C, et al. Legionnaires? disease pseudoepidemic due to 1. falsely positive urine antigen test results. J Clin Microbiol. 2014;52(6):2279-2280. doi:10.1128/JCM.00493-14
1. Briones ML, Blanquer J, Ferrando D, Blasco ML, Gimeno C, Marín J. Assessment of analysis of urinary pneumococcal antigen by immunochromatography for etiologic diagnosis of community-acquired pneumonia in adults. Clin Vaccine Immunol. 2006;13(10):1092-1097. doi:10.1128/CVI.00090-06
1. Athlin S, Iversen A, Özenci V. Comparison of the ImmuView and the BinaxNOW antigen tests in detection of Streptococcus pneumoniae and Legionella pneumophila in urine. Eur J Clin Microbiol Infect Dis. 2017;36(10):1933-1938. doi:10.1007/s10096-017-3016-6