3M Rapid Detection RSV Test

Transcription

1 3M Rapid Detection RSV Test IVD FOR IN VITRO DIAGNOSTIC USE INTENDED USE The 3M Rapid Detection RSV Test is a qualitative, immunochromatographic test for the detection of Respiratory Syncytial Virus (RSV) F-protein antigens in nasal wash/aspirate, nasopharyngeal aspirate and nasopharyngeal swab samples. It is an in vitro diagnostic assay that aids in the rapid diagnosis of RSV infections in symptomatic patients 21 years of age and younger. A negative test is presumptive and it is recommended that all negative results be confirmed by cell culture or direct specimen fluorescence assay (DSFA). Negative results do not preclude RSV infection and should not be used as the sole basis for treatment or other management decisions. The test is intended for professional use. SUMMARY AND EXPLANATION OF THE TEST Respiratory syncytial virus (RSV) is the most common cause of severe lower respiratory tract disease among infants and young children. Persons at increased risk for severe disease or death include premature infants, older adults, and persons of any age with compromised respiratory, cardiac, or immune systems 1,2. RSV is transmitted from person to person via close contact, droplets, or fomites. In temperate climates, peak RSV activity typically occurs during the winter. However, year-to-year national and regional variability in the RSV season onset and offset occurs in the United States 3. Appropriately timed diagnostic tests can provide data that indicate when the RSV season begins nationally and regionally, information that has been critical in determining when to begin RSV prophylaxis for infants and children at high risk for infection 3. Currently available methods for diagnosis of RSV infections include cell culture; serology; and direct examination of respiratory secretions. These include electron microscopy, indirect and direct immunofluorescent antibody tests (DFA), enzyme-linked immunoassays (EIAs), and nucleic acid amplification (e.g. polymerase chain reaction (PCR)) 4. The 3M Rapid Detection RSV Test is a rapid lateral-flow immunoassay that can be utilized to aid in the diagnosis of RSV infection in patients 21 years of age and younger. PRINCIPLES OF THE TEST The 3M Rapid Detection RSV Test is a qualitative immunochromatographic test that utilizes the 3M Rapid Detection Reader for the detection of RSV F-protein antigens in nasal wash/aspirate, nasopharyngeal aspirate (NP aspirate) and nasopharyngeal swab (NP swab) samples. A wash/aspirate or swab sample is added to the Sample Buffer. This sample is then mixed with the Assay Tip containing fluorescent-dyed particles conjugated to specific antibodies and applied into the sample well of the Test Cartridge. The sample migrates along the strip. Fluorescent-dyed particles coated with anti-rsv antibodies bind to RSV F-protein antigens, if present in the sample. As the sample migrates along the strip, RSV-bound particles are captured at the detection zone. Excess fluorescent-dyed particles are captured at the internal standard zone. The Reader then measures the amount of fluorescence emitted by the complexes at the detection zone and at the internal standard zone. The instrument calculates a ratio (RAMP Ratio) of the detection zone fluorescence reading to the internal standard zone fluorescence reading. The Reader then compares these ratios to pre-defined threshold limits to determine a positive or negative result for RSV in the tested sample. Page 1 of 14

2 REAGENTS AND MATERIALS PROVIDED The 3M Rapid Detection RSV Test kit contains all the reagents necessary for the detection of RSV F-protein antigens in nasal wash/aspirate, NP aspirate and NP swab samples. The 3M Rapid Detection RSV Test kit should be stored at 15-30ºC. Do not freeze. Kit contents are stable until the expiration date printed on the box. Test Cartridge 25 Individually foil pouched with desiccant. Packaged with Assay Tip. Contains membrane coated with monoclonal antibodies targeting the RSV F-protein. Assay Tip 25 Packaged with Test Cartridge. Polypropylene tips containing fluorescent particles coated with monoclonal antibodies targeting the RSV F-proteins (contains azide as a preservative). Sample Buffer Vial 25 Containing phosphate buffer, animal protein, surfactant and ProClin 300/ ProClin 950 as preservatives. (contains 150 µl) Disposable Dropper 25 Fixed-volume Disposable Bulb Dropper used to transfer sample to the test Sample Buffer vial. (delivers 150 µl) Transfer Device 1 Fixed volume device used to transfer sample mixed with sample buffer into Test Cartridge. (delivers 75 µl) Control Swab 1 Individually foil pouched with desiccant. Swab coated with non-infectious RSV antigen (gamma irradiated). Negative Control Swab 1 Individually foil pouched with desiccant. Swab coated with non-infectious protein stabilizer solution. Lot Card 1 Stored in anti-static protective pouch. Package Insert 1 Procedure Card 1 MATERIALS REQUIRED BUT NOT PROVIDED 3M Rapid Detection Reader Viral Transport Media for swab sample elution. Refer to the SPECIMEN COLLECTION AND HANDLING section. Specimen collection materials (NP swab, wash/aspirate or NP aspirate container) Disposable gloves Page 2 of 14

3 OPTIONAL MATERIALS 3M Rapid Detection RSV Control Swabs, REF 3966, (5, 5 Negative). Refer to EXTERNAL QUALITY CONTROL SWAB PROCEDURE. WARNINGS AND PRECAUTIONS 1. For in vitro diagnostic use. 2. The device contains material of animal origin and should be handled as a potential biohazard. 3. Do not use the Sample Buffer if it is turbid. Turbidity may be a sign of microbial contamination. 4. Read the entire Package Insert prior to use. Directions should be read and followed carefully, or invalid or erroneous results may occur. 5. RSV virus antigens are relatively unstable. Care should be taken to store samples as indicated in SPECIMEN COLLECTION AND PREPARATION. 6. ProClin is a potential skin sensitizer. Avoid spilling or splashing reagents containing ProClin on skin or clothing. In case of contact, thoroughly flush with water. 7. Once opened, Test Cartridges, Assay Tips and Control Swabs must be used or discarded within 60 minutes. 8. Discard and do not use any visibly damaged cartridges, or the contents of any Test Cartridge, Assay Tip, or Control Swab pouch with a damaged seal. Do not use Test Cartridge, Assay Tip, or Control Swab if desiccant is missing from pouch. 9. Do not use kit contents beyond their expiration date. 10. Do not interchange or mix test components from different kit lots. 11. Use the provided Assay Tip and other test components as directed in the Test Procedure. 12. Assay Tips included in the 3M Rapid Detection RSV Test contain 0.02% (w/v) of sodium azide. Sodium azide is a skin irritant. Avoid skin contact. Disposal of reagents containing sodium azide into drains consisting of lead or copper plumbing can result in the formation of explosive metal oxides. Eliminate build-up of oxides by flushing drains with large volumes of water during disposal. 13. Control Swabs contain inactivated RSV virus and should be handled as if potentially infectious. 14. The 3M Rapid Detection RSV Test Cartridge, Disposable Dropper, Assay Tip, Control Swabs and Sample Buffer Vial should be discarded after single-use. Do not reuse. 15. Do not insert a Test Cartridge that is wet into the Reader, as this may cause contamination or damage to the Reader. If wet, wipe spills before inserting cartridge into Reader. 16. Some patient specimens contain infectious agents; therefore all patient specimens should be handled and discarded as if they are biologically hazardous. Use appropriate precautions in the collection, handling, storage and disposal of clinical specimens, including swabs, and used kit contents. Refer to institutional guidelines for biological waste management. 17. All respiratory samples must be mixed thoroughly before testing, regardless of consistency, to ensure a representative sample prior to testing. 18. Store this product at 15-30ºC when not in use. Do not freeze. Page 3 of 14

4 SPECIMEN COLLECTION AND PREPARATION NOTE: Proper specimen collection, storage, and transport are critical to the performance of this test. Inadequate or inappropriate specimen collection, storage, and/or transport could potentially yield false negative results. Appropriate training in specimen collection is highly recommended to ensure specimen quality. Warning: Specimens contaminated with blood (>2% v/v) may interfere with test results. Do not use specimens that are visibly contaminated with blood. For best test results specimens should be tested as soon as possible. If samples are not tested within 4 hours, they should be stored at 2-8ºC. Specimens may be held up to 72 hours at 2-8ºC prior to testing. Transport Media The following Transport Media are recommended and have been tested and shown not to interfere with the performance of the test. Copan Universal Transport Media (UTM) Remel M4 Media Remel M4-RT Media Remel M5 Media Starplex Transport Media Normal Saline Solution Phosphate Buffered Saline (PBS) Solution Specimen Types and Collection Acceptable specimens for testing with the 3M Rapid Detection RSV Test include nasopharyngeal swab, nasal wash/aspirate, and nasopharyngeal aspirate. The following specimen collection methods are recommended 5. Nasopharyngeal Swab Sample For NP swabs, use sterile foam, polyester, nylon, or rayon swabs. Calcium alginate swabs are not recommended 6. Insert swab into one nostril. Press swab tip on the mucosal surface of the mid-interior portion of the inferior turbinate, and rub the swab tip several times across the mucosal surface to loosen and collect cellular material. Withdraw swab; immediately insert swab into sterile vial of 2-3 ml of recommended transport media by vigorously rotating the swab in the liquid. Nasal Wash: Bulb Method Suction 3-5 ml saline into a new sterile bulb. Insert bulb into one nostril until nostril is occluded. Instill saline into nostril with one squeeze of the bulb and immediately release bulb to collect recoverable nasal secretion. Empty bulb into suitable dry, sterile specimen container or one containing transport media. Nasal Wash: Syringe Method Fill syringe with saline; attach tubing to the syringe tip. Quickly instill saline into nostril. Aspirate the recoverable nasal specimen. Recovery must occur immediately, as the instilled fluid will rapidly drain. Alternatively, patients may tilt head forward to allow specimen to drain into suitable sterile container. If aspirated, inject aspirated specimen from syringe into suitable dry, sterile specimen container or one containing transport media. Page 4 of 14

5 Nasopharyngeal Aspirate Attach mucus trap to suction pump and catheter, leaving wrapper on suction catheter; turn on suction and adjust to suggested pressure. Without applying suction, insert catheter into the nose, directed posteriorly toward the opening of the external ear. Apply suction: using a rotating movement, slowly withdraw the catheter. Catheter should be in nasopharynx no longer than 10 seconds. Hold trap upright to prevent secretions from going into pump. Rinse catheter with saline or transport media (if necessary), disconnect suction, connect tubing to arm of mucus trap to seal. CALIBRATION PROCEDURE Each 3M Rapid Detection RSV Test kit includes a Lot Card that is individually packaged in an anti-static pouch. The Lot Card provides information specific to the kit lot, including lot number, expiration date and threshold information. For further details on loading lot-specific information, see the 3M Rapid Detection Reader Operator Manual. No additional calibration, beyond the insertion of the Lot Card is necessary. This operation is required only once per test kit lot. WARNING: Avoid touching the contacts at the end of the Lot Card. For additional information please refer to the 3M Rapid Detection Reader Operator Manual. EXTERNAL QUALITY CONTROL SWAB PROCEDURE NOTE: Do not extract the Control Swabs directly into the Sample Buffer Vials. The and Negative Control Swabs are designed to be eluted into viral transport media used for patient samples. 1. Obtain a vial of viral transport media (2-3 ml). 2. Open and remove Control Swab (positive or negative) from pouch. 3. Insert head of Control Swab into transport media. 4. Rotate Control Swab in liquid vigorously for 10 seconds. 5. Press Control Swab against the side of the vial as it is removed. 6. Discard Control Swab in appropriate container. 7. Follow the TEST PROCEDURE steps to test the prepared liquid quality control (LQC) sample. TEST PROCEDURE This test should be performed by qualified personnel per local, state or federal regulations or accrediting agency requirements. NOTE: Samples must be at room temperature prior to testing. If stored at 2-8 C, allow samples to equilibrate to room temperature for at least 15 minutes. You will need the Transfer Device and for each sample being tested, one Disposable Dropper, one Sample Buffer Vial and a pouch containing one Test Cartridge and one Assay Tip. 1. Prepare Reader for Test Cartridge. Refer to the 3M Rapid Detection Reader Operator Manual for detailed instructions on Starting an Assay or Running LQC. 2. Ensure that the specimen is well mixed. 3. Place Sample Buffer Vial upright on a clean, dry, level surface or in a rack. Ensure all liquid is at the bottom of the vial and remove the lid. 4. Firmly squeeze the top bulb on a new Disposable Dropper. Place the tip of the Dropper into the specimen and release the bulb to fill the Dropper with 150 µl of the specimen. 5. Ensure there are no air bubbles in lower stem of Dropper. Insert lower stem of Dropper into Sample Buffer and squeeze top bulb to transfer sample into Sample Buffer. Dispose of used Dropper. Page 5 of 14

6 6. Open pouch containing Test Cartridge and Assay Tip. Place Test Cartridge on clean, dry, level surface. 7. Firmly attach Assay Tip to the supplied Transfer Device. 8. Using the Transfer Device (75 μl), insert Assay Tip into Sample Buffer close to, but not touching bottom of vial. Do not press against the bottom of vial as this may block tip. 9. Mix sample slowly by pressing and releasing Transfer Device plunger 10 times (2 seconds per cycle), taking care each time to eject entire sample into Sample Buffer and to draw only liquid and no air into Assay Tip. This will prevent foaming. 10. Position filled Assay Tip directly over sample well of Test Cartridge and fully depress plunger to dispense mixed sample into sample well. (Disregard any remaining droplet in Assay Tip). Dispose of used Assay Tip and Sample Buffer according to local biohazard procedures. 11. Immediately (within 30 seconds) insert Test Cartridge into Reader, and press until firm resistance is felt. Delay will lead to an Error message. 12. Do not insert a Test Cartridge that is wet into the Reader, as this may cause contamination or damage to the Reader. See the 3M Rapid Detection Reader Operator Manual for appropriate cleaning procedure. 13. Test result is complete approximately 15 minutes from Test Cartridge insertion. 14. Record the results. 15. Remove the used Test Cartridge when test is completed and results have been recorded. Dispose of used Test Cartridge according to local biohazard procedures. For additional information on the operation of the Reader, please refer to the 3M Rapid Detection Reader Operator Manual. Refer to the 3M Rapid Detection Reader Operator Manual Troubleshooting section for full description of all Messages. If repeated tests give unexpected or inconsistent results, contact the 3M Health Care Helpline ( ) for assistance; or outside the U.S. and Canada, contact your local 3M subsidiary for assistance. INTERPRETATION OF RESULTS Negative: The Reader screen will display RSV NEG. : The Reader screen will display RSV POS. Error Message: The Reader screen will display an Error message. Refer to the 3M Rapid Detection Reader Operator Manual for detailed Error message information. If an Error message is displayed, the results are invalid, repeat the test using fresh test components. If any of the following Error messages are displayed and the sample is viscous (mucoid), add an approximately equal volume transport media to the remaining original sample, mix by inversion and repeat the test on this diluted sample: Error Message Port Display Sample Error 2 Fail Low Signal LOW High Background HRB NOTE: Diluting the sample may lower the overall test sensitivity. Page 6 of 14

7 REPORTING OF RESULTS Result Recommendation RSV NEG Report negative test results as RSV virus antigen not detected. This result does not exclude RSV infection. A negative test is presumptive and it is recommended that these results be confirmed by cell culture or DSFA. RSV POS Report positive test results as for RSV virus antigen. This result does not rule out co-infections with other pathogens. QUALITY CONTROL Refer to the 3M Rapid Detection Reader Operator Manual for full details on quality control measures and troubleshooting. System Quality Control The instrument has error checking and self-diagnostic functions that assure procedural control. These include algorithms and measurements used to confirm acceptable operator technique, sample handling and test performance. If a problem is detected, a message is displayed. Contact the 3M Health Care Helpline ( ) for assistance, or outside the U.S. and Canada, contact your local 3M Subsidiary for assistance. Valid results are displayed only after all performance requirements have been met. Procedural Controls The 3M Rapid Detection RSV Test has built-in (procedural) controls. Each Test Cartridge has an internal standard zone that is scanned as part of the test protocol to ensure proper sample flow. Control limits for each lot of Test Cartridges are established during the manufacturing process and are incorporated in the specific Lot Card. If a procedural control result does not meet specifications, the sample result is not reported and an Error message is displayed. External and Negative Control Swabs See section on EXTERNAL QUALITY CONTROL SWAB PROCEDURE for instructions on performing these control tests. The reactivity of each new lot and of each new shipment of 3M Rapid Detection RSV Test kit should be verified on receipt using external and Negative Quality Control Swabs. Additional external positive and negative control testing may be required in accordance with applicable local, state and/or federal guidelines or accreditation requirements and each facility s standard Quality Control procedures. Additional and Negative Control Swabs (REF 3966) are available from 3M Health Care. These controls are used to monitor reagent reactivity. Failure of the controls to produce the expected results can mean that one of the reagents or components is no longer reactive at the time of use, the test was not performed correctly or that reagents or samples were not added. If the control tests do not produce the expected result, repeat with fresh Control Swabs, samples and fresh test components. Whenever control results fail, controls and samples should be retested. Do not perform patient tests or report patient test results if the control tests do not produce the expected results. For assistance in troubleshooting contact the 3M Health Care Helpline ( ), or outside the U.S. and Canada, contact your local 3M Subsidiary. and negative tests are described in the section on INTERPRETATION OF RESULTS. Control Swab, containing RSV antigens, is expected to display RSV Page 7 of 14

8 POS. Negative Control Swab is expected to display RSV NEG. It is a control failure when the Control Swabs do not produce expected results. The external and Negative Control Swabs manufactured for this test are prepared in a simulated sample matrix, which may not mimic test specimens. If external control materials that are identical in composition to test specimens are preferred, the user can prepare those by using known positive and negative specimens in saline solution according to the SPECIMEN COLLECTION AND PREPARATION section. For further information regarding matrix effects refer to The Clinical Laboratory Standards Institute (CLSI) guideline on matrix effects EP14-A2. 7 LIMITATIONS 1. The 3M Rapid Detection RSV Test can only be used with the 3M Rapid Detection Reader. 2. and negative predictive values are highly dependent on prevalence. False negative test results are more likely during peak activity when prevalence of disease is high. False positive test results are more likely during periods of low RSV activity when prevalence is moderate to low. 3. Tests using monoclonal antibodies may fail to detect, or detect with less sensitivity, RSV viruses that have undergone minor amino acid changes in the target epitope region. 4. Sample types, swabs or viral transport media other than those listed have not been evaluated and should not be used. 5. The 3M Rapid Detection RSV Test detects both viable and non-viable RSV viruses. Test performance depends on antigen load and may not correlate with cell culture or DSFA results. 6. The test results are to be used in conjunction with information available from the patient s clinical evaluation and other diagnostic procedures. 7. A test result that is inconsistent with the clinical signs and symptoms should be interpreted with caution. 8. Specimens contaminated with whole blood >2% v/v (visibly bloody samples) may interfere with test results and should not be used. 9. Viscous (mucoid) samples may require dilution to obtain valid results. This dilution may lower the overall test sensitivity. 10. Inadequate or improper sample handling/transport may yield a false negative result. 11. A negative test is presumptive and it is recommended these results be confirmed by cell culture or DSFA. Negative results do not preclude RSV infection and should not be used as the sole basis for treatment or other management decisions. 12. Analytical studies were conducted using RSV A samples only. Potential interference with RSV B was not assessed M Rapid Detection RSV Test potential cross-reactivity with Chlamydophila pneumoniae has not been determined. EXPECTED VALUES The prevalence of RSV varies each year with epidemics occurring during the fall and winter months in the US. Variables that affect the rate of positivity observed in respiratory testing include: the efficiency and timing of specimen collection, handling and transport of the specimen, the time of year, age of the patient, and local disease prevalence. The prevalence observed with culture and DSFA during the clinical study (November 2008 February 2009) was 33.9% (443/1306). During the previous U.S. respiratory season (October 2007 March 2008), the prevalence of RSV was 15.9% 8. Page 8 of 14

9 PERFORMANCE CHARACTERISTICS CLINICAL STUDIES The performance of the 3M Rapid Detection RSV Test was compared to cell culture and DSFA in a prospective study conducted as part of a multi-center trial during the RSV season. Eight (8) independent laboratories located in distinct geographical regions across the United States (NY, MO(2), MD, OH(2), MA, AR) evaluated the 3M Rapid Detection RSV Test in parallel with cell culture / DSFA. Testing staff included both laboratory and non-laboratory personnel, and two sites (MO, AR) also performed testing in point-of-care settings (near patient). The sites included an Emergency Department and Pediatric Testing Unit.. One thousand, two hundred and seventy nine (1279) fresh specimens were collected from pediatric patients 21 years of age and younger. Of these specimens, valid results were obtained from 1140, with an approximately equal mix of male and female patients. During the clinical study, if either the cell culture or DSFA result was positive for a patient sample, it was determined to be a positive reference result. A reference result was deemed negative only if both, culture and DSFA, were negative. Prospective Study Table 1. Results with Prospective Nasopharyngeal Swab Samples Culture / DSFA NP Swab 3M Rapid Detection RSV Negative Total Negative 7* Total % CI Sensitivity 88.2% Specificity 97.4% * 1 was positive by the 3M Rapid Detection RSV Test and by the Prodesse ProFlu RT-PCR analysis. Table 2. Results with Prospective Nasopharyngeal Aspirate Samples Culture / DSFA NP Aspirate 3M Rapid Detection RSV Negative Total * 197 Negative 15** Total % CI Sensitivity 85.3% Specificity 95.1% * 8 were negative by the 3M Rapid Detection RSV Test and by an investigational RT-PCR. ** 3 were positive by the 3M Rapid Detection RSV Test and by an investigational RT-PCR. Page 9 of 14

10 Table 3. Results with Prospective Nasal Wash / Aspirate Samples Culture / DSFA Nasal Wash / Aspirate 3M Rapid Detection RSV Negative Total 98 11* 109 Negative 9** Total % CI Sensitivity 89.9% Specificity 93.4% * 1 was negative by the 3M Rapid Detection RSV Test and by an investigational RT-PCR. ** 1 was positive by the 3M Rapid Detection RSV Test and by an investigational RT-PCR. PRECISION AND REPRODUCIBILITY The precision and reproducibility of the 3M Rapid Detection RSV Test was evaluated using a panel of 3 simulated samples that included high negative, low positive (at LoD) and moderate positive (2 x LoD) RSV samples. The RSV strain used to prepare the samples was RSV A (A-2) strain. To evaluate reproducibility, multiple operators at multiple sites tested each of the three precision samples on 5 different days. To evaluate precision, at one site, operators tested each of the three precision samples on 12 different days. There was 99.2% agreement with the expected test results for all specimens tested, with no differences within run (same operator on same day) between run, operators or sites. Table 4. Precision and Reproducibility Study Results RSV RSV Panel High Low Member ID Negative Viral Titer (TCID 50/mL) RSV Moderate Total Agreement All (%) Site 1 Site 2 Site 3 Agreement 30/30 30/30 30/30 Mean RAMP Ratio* Value % CV 11% 11% 9% Agreement 30/30 29/30 30/30 Mean RAMP Ratio Value % CV 13% 14% 18% Agreement 71/72 71/72 72/72 Mean RAMP Ratio Value % CV 17% 12% 11% Total Agreement 131/132 (99.2%) 130/132 (98.5%) 132/132 (100%) 90/90 (100%) 89/90 (98.9%) 214/216 (99.1%) 393/396 (99.2%) 95% CI % % % % Overall Mean RAMP Ratio Overall % CV 16% 13% 14% *RAMP Ratios are for reference only. These values are not reported by the instrument. Page 10 of 14

11 INTERFERENCE Whole blood and a number of other potentially interfering substances (medications and over the counter (OTC) products) that may be present naturally or artificially introduced in the nasal cavity or nasopharynx were evaluated in the 3M Rapid Detection RSV Test. The substances were added to a simulated negative sample (0.85% saline), an RSV LoD positive sample and an RSV 2x LoD positive sample and tested in the 3M Rapid Detection RSV Test. The RSV strain used to prepare the samples was RSV A (A-2), interference was not evaluated with an RSV B strain.. The following substances were found to have no effect on the test results of the 3M Rapid Detection RSV Test when present in positive and negative RSV simulated respiratory samples at the concentrations indicated: Whole blood (2% v/v), mucin (1% w/v), Scope Mouthwash (40% v/v), Good and Kind Mouthwash (40% v/v), Cepacol Mouthwash (40% v/v), Benzocain as Cepacol Throat Drop (30% w/v), Menthol as Fisherman s Friend Throat Drop (30% w/v), Budesonide as Rhinocort Nasal Spray (15% v/v), Triamcinolone as Nasocort (15%), Fluticasone Furoate as Flonase Nasal Spray (30% v/v), Mometasone Furoate as Nasonex Nasal Spray (15% v/v), 4- Acetamidophenol as Tylenol (20 mg/ml), Acetylsalicylic acid as Aspirin (30 mg/ml), Chloropheniramine as Chlor-Tripolon Tablet (10 mg/ml), Diphenylhydramine as Benadryl Allergy Tablet (5 mg/ml), Dextromethorphan as Delsym DM Cough Syrup (2 mg/ml), Guaiacol Glycerol Ether as Robitussin Syrup (40 mg/ml), Phenylpropanolamine HCl (pure) (40 mg/ml), Oxymetazoline HCl as Afrin Nasal Spray (0.05% v/v), Phenylephrine HCl as Neo- Synephrine Nasal Spray (20 mg/ml), sodium chloride with preservatives as Otrivin Saline (1.4% w/v), Ribavirin as Rebetol (100 ng/ml), Zanamivir as Relenza (20 mg/ml), Rimantadine HCl (500 mg/ml), and Oseltamivir Phosphate as Tamiflu (50 mg/ml). ANALYTICAL SENSITIVITY AND REACTIVITY The analytical sensitivity (limit of detection or LoD) and reactivity of the 3M Rapid Detection RSV Test was determined using quantified (TCID 50 /ml) cultures of 2 RSV subtype A and 4 RSV subtype B strains serially diluted in either viral transport media (VTM) to simulate a swab sample type or saline solution to simulate a wash/aspirate sample type. The RSV strains were re-titered prior to testing. Each viral strain was tested in replicates of 20 per concentration of virus. Analytical sensitivity (LoD) was defined as the lowest concentration, at which 95% of all replicates tested positive. Table 5. Analytical Sensitivity and Reactivity Study Results LoD RSV Sample Concentration Result (TCID 50 /ml) 3 100% RSV A-Long in Saline 6.5x % RSV A-Long in VTM 2.5x % RSV A (A-2) in Saline 1.0x % RSV A (A-2) in VTM 1.2x % RSV B CH93(18)-18 in Saline 3.5x % RSV B CH93(18)-18 in VTM 3.5x % RSV B Wash/18537/62 in Saline 7.0x10 Page 11 of 14

12 RSV Sample LoD Concentration (TCID 50 /ml) Result 3 100% RSV B Wash/18537/62 in VTM 6.0x % RSV B WV/14617/85 in Saline 5.0x % RSV B WV/14617/85 in VTM 2.5x % RSV B9320 in ZMC matrix* > 1.7x10 *Culture supplied by Zeptometrix Corporation in proprietary matrix CROSS REACTIVITY The analytical specificity of the 3M Rapid Detection RSV Test was evaluated by testing a panel consisting of 16 viruses and 17 bacteria that may be present in the nasal cavity or nasopharynx. Bacterial and viral isolates were tested at the concentrations listed below. The bacterial and viral organisms were re-grown and re-titered prior to cross-reactivity testing. None of the organisms tested gave a positive result in the 3M Rapid Detection RSV Test. Table 6. Cross Reactivity Study Results Strain/Isolate Concentration RSV result Adenovirus, Type TCID 50 /ml Negative Adenovirus, Type 7a 10 5 TCID 50 /ml Negative Human coronavirus, Strain OC TCID 50 /ml Negative Human coronavirus, Strain 229E 10 5 TCID 50 /ml Negative Cytomegalovirus 10 5 TCID 50 /ml Negative Enterovirus, Type TCID 50 /ml Negative Epstein Barr Virus 10 5 TCID 50 /ml Negative Human Parainfluenza, Type TCID 50 /ml Negative Human Parainfluenza, Type TCID 50 /ml Negative Human Parainfluenza, Type TCID 50 /ml Negative Influenza A, Brisbane/10/ EID 50 /ml Negative Influenza B, Ohio/01/ TCID 50 /ml Negative Measles 10 5 TCID 50 /ml Negative Human metapneumovirus 10 5 TCID 50 /ml Negative Mumps virus 10 5 TCID 50 /ml Negative Human Rhinovirus, Strain 1A 10 5 TCID 50 /ml Negative Bordetella pertussis 10 6 cfu/ml Negative Corynebacterium Sp cfu/ml Negative Escherichia coli 10 6 cfu/ml Negative Haemophilus influenzae 10 6 cfu/ml Negative Lactobacillus casei 10 6 cfu/ml Negative Legionella pneumophila 10 6 cfu/ml Negative Moraxella catarrhalis 10 6 cfu/ml Negative Mycobacterium tuberculosis, Avirulent 10 6 cfu/ml Negative Mycoplasma pneumoniae 10 6 cfu/ml Negative Neisseria meningitidis 10 6 cfu/ml Negative Page 12 of 14

13 Strain/Isolate Concentration RSV result Neisseria sicca 10 6 cfu/ml Negative Pseudomonas aeruginosa 10 6 cfu/ml Negative Staphylococcus aureus, Strain cfu/ml Negative Staphylococcus epidermidis, Strain cfu/ml Negative Streptococcus pneumoniae 10 6 cfu/ml Negative Streptococcus pyogenes 10 6 cfu/ml Negative Streptococcus salivarius 10 6 cfu/ml Negative NOTE: 3M Rapid Detection RSV Test potential cross-reactivity with Chlamydophila pneumoniae has not been determined. ASSISTANCE If you have any questions regarding the use of this product please contact the 3M Health Care Helpline ( ). If outside the U.S. or Canada, contact your local 3M Subsidiary for assistance. REFERENCES 1 Falsey AR, Hennessey PA, Formica MA, Cox C, Walsh EE. Respiratory syncytial virus infection in elderly and high-risk adults. N Engl J Med 2005;352: Welliver RC. Review of epidemiology and clinical risk factors for severe respiratory syncytial virus (RSV) infection. J Pediart 2003;143(5 Suppl):S Centers for Disease Control and Prevention Website, Morbidity and Mortality Weekly Report, December 7, 2007 / 56(48); , Brief Report: Respiratory Syncytial Virus Activity --- United States, July November 2007, 13August 2009, 4 Henrickson, KJ and Hall, CB Diagnostic Assays for Respiratory Syncytial Virus Disease (Pediatr Infect Dis J 2007;26: S36-S40) 5 Storch GA (ed): Essentials of Diagnostic Virology 1999; Churchill-Livingstone, New York, NY. 6 Lauer BA, Masters HB. Toxic effect of calcium alginate swabs on Neisseria gonorrhoeae. J Clinical Microbiol Jan;26(1): Clinical and Laboratory Standards Institute. Evaluation of Matrix Effects; Approved Guideline-Second edition. Document EP-14A2. Wayne, PA: Clinical and Laboratory Standards and Institute; Centers for Disease Control and Prevention Website, Morbidity and Mortality Weekly Report, December 19, 2008 / 57(50); , Brief Report: Respiratory Syncytial Virus Activity --- United States, July December 2008, 13August 2009, EXPLANATION OF SYMBOLS Symbol Definition Catalog Number Batch Code Use By Page 13 of 14

14 Symbol Definition Do Not Reuse Consult Instructions for Use Caution, Consult Accompanying Documents In vitro diagnostic medical device Manufacturer Contains sufficient for <n> tests Temperature limitation control Negative control Authorized European Representative REF M Rapid Detection RSV Test 25 tests IVD Made in Canada (Control Swabs made in Italy) for 3M Health Care St. Paul, MN U.S.A. 3M Health Care D Neuss, Germany 3M is a registered trademark of 3M. RAMP Technology is a registered trademark of Response Biomedical Corporation. ProClin is a registered trademark of Rohm and Haas Company. Other trademarks are the property of their respective owners. Copyright 2010, 3M. All rights reserved [MAR 2010] Page 14 of 14