Principle of Test NOTE: Materials Provided
2
Intended Use The PIFA PLUSS™ PF4 Rapid Assay (PIFA PLUSS ™ PF4) is a qualitative in vitro diagnostic device designed for the detection of antibodies to Platelet Factor 4 (PF4) sensitized microspheres. These antibodies are found in some patients undergoing heparin therapy. Summary and Explanation Summary The risk of heparin-induced thrombocytopenia is greatly increased in patients with recent exposure to heparin. Immune-mediated heparin-induced thrombocytopenia (HIT) is often caused by platelet-activating antibodies that recognize complexes of Heparin/PF4. As a result, antibody detection is rapidly becoming a standard of care in hematology and cardiology. The majority of laboratory tests for HIT are classified by CLIA as high complexity, take many hours to perform, and often provide confirmation of HIT or HIT and thrombosis (HITT) after the symptoms are seen in a patient. As a result, there is a need for an easily performed, rapid test that helps clinicians identify and treat patients at risk for HIT or thrombosis. Explanation HIT is caused by heparin-dependent antibodies (HIT Antibodies) formed to the heparin/PF4 complex. HIT antibodies are most frequently induced by Unfractionated Heparin (UFH) use following Cardiopulmonary Bypass Surgery (50%) and major Orthopedic Surgery (15%); ultimately HIT is observed in 1-to- 5% of patients treated with UFH (1) . HIT antibodies are initially formed when a patient has been on heparin therapy for five or more days. An immune response to a heparin dose may be observed sooner (1-to-2 days) if the patient has had previous exposure to heparin. The hallmark symptoms of HIT are a drastic fall in platelet count and thrombosis. Other symptoms may include cutaneous reactions, from a simple allergic reaction to lesions or necrosis. There are two categories of laboratory methods used to identify HIT antibodies: Functional/Activation Assays and Antigen Assays/Immunoassays. Functional Assays include the C-14 Serotonin Release Assay (SRA) and the Platelet Aggregation Test (PAT); these assays assess the platelet-activating potential of HIT antibodies. The Antigen Assay class consists of the solid-phase Enzyme-Linked Immunsorbent Assay (ELISA) and the liquid-phase Particle ImmunoFiltration Assay (PIFA ® ) which determine the presence or absence of HIT Antibodies. SRAs, PATs and ELISAs are CLIA-classified as high complexity, take many hours to perform or are often completed sporadically in “batch” or out-sourced. (2) Results from these tests frequently provide clinicians with retrospective patient data that cannot be factored into time-sensitive anticoagulation decisions. The PIFA PLUSS™ PF4 is a rapid manual assay that can easily be performed in minutes, in-house and is CLIA-classified as moderate complexity. Like the PIFA Heparin/PF4 Rapid Assay (PIFA HPF4), the PIFA PLUSS™ PF4 Assay detects antibodies to complexes of heparin/PF4. While the PIFA HPF4 utilizes serum obtained from Red Top blood collection tubes, the first-in-class PIFA PLUSS™ PF4 assay utilizes ABI’s patented seraSTAT™ Blood Cell Separator to extract and deliver a measured volume of the liquid fraction of Whole Blood into the PIFA PLUSS™ PF4 device. Principle of Test The PIFA PLUSS™ PF4 is based upon principles of the Particle ImmunoFiltration Assay (PIFA ® ). BLUE, microparticle-based reagents, coated with purified Platelet Factor 4 (PF4) protein derived from platelet-rich plasma obtained from human sources provide the visual signal for the results of the assay. The inability of matrixed microparticles, or the ability of non-matrixed particles, to move through a filtration medium is the measure of the reactivity/nonreactivity of the test sample. The PIFA PLUSS™ PF4 consists of two components - a MiniReactor device and the seraSTAT™ Blood Cell Separator (seraSTAT). The MiniReactor features a push button reagent dispensing system, referred to as the TOWER, that contains BLUE, microparticle-based reaction reagents (reagent). The microparticles are coated with purified PF4 protein, as well as additional enhancing agents designed to promote rapid aggregation or matrix formation of the microparticles in the presence of HIT antibodies in the test sample. The MiniReactor also contains a membrane filtration system, a REAGENT Window, a TEST Result Window, and a device CONTROL Window. The Test Procedure is initiated with 150 µL of the patient’s Whole Blood dispensed onto the seraSTAT™ sample well. The membrane within the seraSTAT™ separates the specimen into liquid and cellular fractions. Once YELLOW reaches the Lower Edge of the membrane, this signals that the measured volume of liquid fraction has been extracted. The seraSTAT™ is then inserted into the Slot in the TOWER. Manual pressure is applied to the TOWER to deliver the prepared specimen into the TOWER’s internal reaction chamber. The YELLOW specimen makes contact with the BLUE, microparticle-based reagent. After a brief, 65-second mixing and reaction step, the mixture is dispensed into the MiniReactor’s membrane filtration system. A Reactive sample will form a matrix of BLUE microparticles that are bound together and trapped within the membrane filter. As a result, only YELLOW from the seraSTAT™-prepared specimen can migrate into the TEST Result Window to signal a Positive result. Conversely, a Non-Reactive sample does not form a matrix of microparticles, allowing the YELLOW specimen and BLUE reagent to form a mixture that passes through the membrane filter; therefore, a BLUE-GREEN color will migrate into the TEST Result Window to indicate a Negative result. Specimen Collection, Preparation, and Storage Collection Only FRESH, anticoagulated blood samples collected with EDTA or Sodium Citrate collection tubes can be used with the PIFA PLUSS™ PF4 assay. No other collection tubes are recommended for use. Do not use frozen and/or thawed specimens. Consult Collection Tube Manufacturer’s instructions for mixing protocols. Preparation The seraSTAT™ blood cell separator must be used to extract the appropriate volume of patient specimen for introduction into the PIFA PLUSS™ PF4 MiniReactor. In accordance with CLSI Guideline H18-A4, “Procedures for the Handling and Processing of Blood Specimens”, patient specimens that remain at room temperature after collection must be introduced to the seraSTAT blood cell separator within a maximum of TWO (2) hours from the draw. (3) . Storage Properly collected anticoagulated whole blood specimens that cannot be tested immediately, should be stored refrigerated (2 to 8°C; 36 to 46°F) for no longer than 24 hours. NOTE: Refrigerated specimens should reach an ambient temperature (18 to 27°C; 64 to 81°F) prior to performing the test. Materials Provided: One (1) PIFA PLUSS™ PF4 Kit (Item #4001101) contains: One (1) PIFA PLUSS™ PF4 Package Insert and Six (6) PIFA PLUSS PF4 Assays packaged in individual sealed pouches. Each sealed pouch contains the following components: One (1) PIFA PLUSS™ PF4 MiniReactor Device One (1) seraSTAT™ Blood Cell Separator Materials Required But Not Provided Timing Device Pipettor capable of delivering 150µl and disposable tips Positive and Negative Controls (see Quality Control) Quality Control The PIFA PLUSS™ PF4 is a single-use/unit use test system. Akers Biosciences, Inc. recommends that the laboratory analyzes a sample representative of the lot for Quality Control. It is also recommended that a positive and negative control be utilized in the following circumstances: 1) during the initial use of a new lot of PIFA PLUSS™ PF4 devices, and 2) after running 100 tests of the same lot of devices. The PIFA PLUSS™ PF4 contains an internal device control with each test run. The appearance of RED in the CONTROL Window indicates that the device has functioned as designed. If RED does not develop in the CONTROL Window within a maximum of 10 minutes after performing the test procedure, the test result is considered invalid. The appearance of YELLOW or BLUE-GREEN in the TEST Result window indicates that a sufficient sample has been introduced into the reaction mixture. Controls should be assayed as required by your laboratory’s standard, quality control procedures using the same test procedure for patient specimens. Use only confirmed Heparin/PF4 antibody positive and negative specimens. Control specimens may be sera or plasma. If sera, the specimens MUST have originated from whole blood samples collected in Plain, Glass Red Top tubes OR Plastic, Red Top Tubes, with Clot Activator; no other collection tubes are recommended for use, including gel serum separation tubes (SST). If plasma, the specimens MUST have originated from anticoagulated blood samples drawn in EDTA or Sodium Citrate collection tubes. Storage Conditions PIFA PLUSS™ PF4 assays must be stored refrigerated (2 to 8°C (36 to 46°F)). Devices that have been removed from refrigeration, left in their sealed pouches, and stored at an ambient temperature (18 to 27°C; 64 to 81°F) for a maximum of 8 hours, may be returned to refrigeration for future use. This cycle can be repeated for a maximum room temperature exposure of 32 hours. Please note: • Do not freeze tests. • If the test is frozen upon receipt, results will be invalid. • Do not use any tests beyond their expiration date. Warnings and Precautions • For in vitro Diagnostic Use. • Do not expose the PIFA PLUSS™ PF4 MiniReactors to temperatures greater than 40°C (104°F) or below 0°C (32°F). • Allow each PIFA ® MiniReactor to warm to an ambient temperature (18 to 27°C; 64 to 81°F), in the individual sealed pouch, for a minimum of 30 minutes prior to performing the test. • PIFA PLUSS™ PF4 must be performed using anticoagulated samples draw with EDTA or Sodium Citrate blood collection tubes. • All patient specimens should be collected using aseptic technique and should be handled in accordance with good laboratory practices. • Frozen, thawed, hemolyzed, icteric, lipemic (of an excessive nature), bacterial contaminated specimens, or controls from other test kits should not be used, and can produce erroneous results. • Inadequate incubation time, incomplete mixing, or improperly performed test procedures can produce erroneous results. • Do not initiate testing with the PIFA PLUSS PF4 assay if the membrane extending from the accompanying seraSTAT™ is torn or creased. • See also “Limitations.” Limitations • The PIFA PLUSS™ PF4 should be used for the qualitative detection of any antibody directed against the PF4 complex, and should be used as a screening test. There may be some antibodies reactive to the Heparin/PF4 complex that are non- reactive with this test. Test result should not be relied upon solely to identify an antibody to the PF4 complex. • A positive test result may be indicative of a Heparin/PF4- related antibody in the test sample. However, the presence of these antibodies does not confirm the diagnosis of HIT or HITT. Therefore, results obtained from the PIFA PLUSS™ PF4 should be interpreted along with clinical findings and/or other serological tests. Performance Characteristics Akers Biosciences, Inc. has conducted a series of evaluations to determine the performance of the PIFA ® Heparin/PF4 Rapid Assay for the detection of antibodies to the Heparin/PF4 complex. Studies were performed by outside laboratories to determine the performance of the PIFA ® Heparin/PF4 Rapid Assay compared to standard laboratory methods using samples originating from field sources. The standard laboratory method was a commercially available ELISA technique. Reproducibility The reproducibility of the PIFA ® Heparin/PF4 Rapid Assay in detecting Heparin/PF4 antibodies was demonstrated by testing 10 aliquots of 5 specimens for inter-day evaluation and 10 aliquots for intra-day evaluation. Reproducibility of the PIFA ® Heparin/PF4 Rapid Assay was determined to be 100% in both studies. References 1 Warkentin TE, Greinacher A, eds. Heparin-induced thrombocytopenia, 3rd ed. New York: Marcel Dekker, Inc. 2004, 53, 205, 271, 295. 2 Daneschvar, H.Leon, MD, Daw, Hamed, MD: How to Interpret and Pursue an Abnormal Complete Blood Cell Count in Adults. Mayo Clinic Proc. 2005;80:1389-1394 2005 Mayo Foundation for Medical Education and Research. 3 CLSI (Clinical and Laboratory Standards Institute) H18-A4: Procedures for the Handling and Processing of Blood Specimens; Approved Guideline -- 4th ed. (ISBN 1-56238-72-3). Specificity and Sensitivity ELISA Positive Negative PIFA ® Positive 21 3 Negative 2 153 • Specificity: 98.1% • Sensitivity: 91.3% • Overall Agreement: 97.2%
Transcript of Principle of Test NOTE: Materials Provided
Intended UseThe PIFA PLUSS™ PF4 Rapid Assay (PIFA PLUSS ™ PF4) is a qualitative in vitro diagnostic device designed for the detection of antibodies to Platelet Factor 4 (PF4) sensitized microspheres. These antibodies are found in some patients undergoing heparin therapy.
Summary and ExplanationSummaryThe risk of heparin-induced thrombocytopenia is greatly increased in patients with recent exposure to heparin. Immune-mediated heparin-induced thrombocytopenia (HIT) is often caused by platelet-activating antibodies that recognize complexes of Heparin/PF4. As a result, antibody detection is rapidly becoming a standard of care in hematology and cardiology. The majority of laboratory tests for HIT are classified by CLIA as high complexity, take many hours to perform, and often provide confirmation of HIT or HIT and thrombosis (HITT) after the symptoms are seen in a patient. As a result, there is a need for an easily performed, rapid test that helps clinicians identify and treat patients at risk for HIT or thrombosis.
ExplanationHIT is caused by heparin-dependent antibodies (HIT Antibodies) formed to the heparin/PF4 complex. HIT antibodies are most frequently induced by Unfractionated Heparin (UFH) use following Cardiopulmonary Bypass Surgery (50%) and major Orthopedic Surgery (15%); ultimately HIT is observed in 1-to-5% of patients treated with UFH(1). HIT antibodies are initially formed when a patient has been on heparin therapy for five or more days. An immune response to a heparin dose may be observed sooner (1-to-2 days) if the patient has had previous exposure to heparin. The hallmark symptoms of HIT are a drastic fall in platelet count and thrombosis. Other symptoms may include cutaneous reactions, from a simple allergic reaction to lesions or necrosis.
There are two categories of laboratory methods used to identify HIT antibodies: Functional/Activation Assays and Antigen Assays/Immunoassays. Functional Assays include the C-14 Serotonin Release Assay (SRA) and the Platelet Aggregation Test (PAT); these assays assess the platelet-activating potential of HIT antibodies. The Antigen Assay class consists of the solid-phase Enzyme-Linked Immunsorbent Assay (ELISA) and the liquid-phase Particle ImmunoFiltration Assay (PIFA®) which determine the presence or absence of HIT Antibodies. SRAs, PATs and ELISAs are CLIA-classified as high complexity, take many hours to perform or are often completed sporadically in “batch” or out-sourced.(2) Results from these tests frequently provide clinicians with retrospective patient data that cannot be factored into time-sensitive anticoagulation decisions. The PIFA PLUSS™ PF4 is a rapid manual assay that can easily be performed in minutes, in-house and is CLIA-classified as moderate complexity. Like the PIFA Heparin/PF4 Rapid Assay (PIFA HPF4), the PIFA PLUSS™ PF4 Assay detects antibodies to complexes of heparin/PF4. While the PIFA HPF4 utilizes serum obtained from Red Top blood collection tubes, the first-in-class PIFA PLUSS™ PF4 assay utilizes ABI’s patented seraSTAT™ Blood Cell Separator to extract and deliver a measured volume of the liquid fraction of Whole Blood into the PIFA PLUSS™ PF4 device.
Principle of TestThe PIFA PLUSS™ PF4 is based upon principles of the Particle ImmunoFiltration Assay (PIFA®). BLUE, microparticle-based reagents, coated with purified Platelet Factor 4 (PF4) protein derived from platelet-rich plasma obtained from human sources provide the visual signal for the results of the assay. The inability of matrixed microparticles, or the ability of non-matrixed particles, to move through a filtration medium is the measure of the reactivity/nonreactivity of the test sample.
The PIFA PLUSS™ PF4 consists of two components - a MiniReactor device and the seraSTAT™ Blood Cell Separator (seraSTAT). The MiniReactor features a push button reagent dispensing system, referred to as the TOWER, that contains BLUE, microparticle-based reaction reagents (reagent). The microparticles are coated with purified PF4 protein, as well as additional enhancing agents designed to promote rapid aggregation or matrix formation of the microparticles in the presence of HIT antibodies in the test sample. The MiniReactor also contains a membrane filtration system, a REAGENT Window, a TEST Result Window, and a device CONTROL Window.
The Test Procedure is initiated with 150 µL of the patient’s Whole Blood dispensed onto the seraSTAT™ sample well.
The membrane within the seraSTAT™ separates the specimen into liquid and cellular fractions. Once YELLOW reaches the Lower Edge of the membrane, this signals that the measured volume of liquid fraction has been extracted. The seraSTAT™ is then inserted into the Slot in the TOWER. Manual pressure is applied to the TOWER to deliver the prepared specimen into the TOWER’s internal reaction chamber.
The YELLOW specimen makes contact with the BLUE, microparticle-based reagent. After a brief, 65-second mixing and reaction step, the mixture is dispensed into the MiniReactor’s membrane filtration system. A Reactive sample will form a matrix of BLUE microparticles that are bound together and trapped within the membrane filter. As a result, only YELLOW from the seraSTAT™-prepared specimen can migrate into the TEST Result Window to signal a Positive result. Conversely, a Non-Reactive sample does not form a matrix of microparticles, allowing the YELLOW specimen and BLUE reagent to form a mixture that passes through the membrane filter; therefore, a BLUE-GREEN color will migrate into the TEST Result Window to indicate a Negative result.
Specimen Collection, Preparation, and StorageCollectionOnly FRESH, anticoagulated blood samples collected with EDTA or Sodium Citrate collection tubes can be used with the PIFA PLUSS™ PF4 assay. No other collection tubes are recommended for use. Do not use frozen and/or thawed specimens. Consult Collection Tube Manufacturer’s instructions for mixing protocols.
PreparationThe seraSTAT™ blood cell separator must be used to extract the appropriate volume of patient specimen for introduction into the PIFA PLUSS™ PF4 MiniReactor. In accordance with CLSI Guideline H18-A4, “Procedures for the Handling and Processing of Blood Specimens”, patient specimens that remain at room temperature after collection must be introduced to the seraSTAT blood cell separator within a maximum of TWO (2) hours from the draw.(3).
StorageProperly collected anticoagulated whole blood specimens that cannot be tested immediately, should be stored refrigerated (2 to 8°C; 36 to 46°F) for no longer than 24 hours.
NOTE: Refrigerated specimens should reach an ambient temperature (18 to 27°C; 64 to 81°F) prior to performing the test.
Materials Provided:One (1) PIFA PLUSS™ PF4 Kit (Item #4001101) contains: One (1) PIFA PLUSS™ PF4 Package Insert and Six (6) PIFA PLUSS PF4 Assays packaged in individual sealed pouches.
Each sealed pouch contains the following components:One (1) PIFA PLUSS™ PF4 MiniReactor DeviceOne (1) seraSTAT™ Blood Cell Separator
Materials Required But Not ProvidedTiming DevicePipettor capable of delivering 150µl and disposable tipsPositive and Negative Controls (see Quality Control)
Quality Control The PIFA PLUSS™ PF4 is a single-use/unit use test system. Akers Biosciences, Inc. recommends that the laboratory analyzes a sample representative of the lot for Quality Control. It is also recommended that a positive and negative control be utilized in the following circumstances:
1) during the initial use of a new lot of PIFA PLUSS™ PF4 devices,and2) after running 100 tests of the same lot of devices.
The PIFA PLUSS™ PF4 contains an internal device control with each test run. The appearance of RED in the CONTROL Window indicates that the device has functioned as designed. If RED does not develop in the CONTROL Window within a maximum of 10 minutes after performing the test procedure, the test result is considered invalid.
The appearance of YELLOW or BLUE-GREEN in the TEST Result window indicates that a sufficient sample has been introduced into the reaction mixture.
Controls should be assayed as required by your laboratory’s standard, quality control procedures using the same test procedure for patient specimens. Use only confirmed Heparin/PF4 antibody positive and negative specimens.
Control specimens may be sera or plasma. If sera, the specimens MUST have originated from whole blood samples collected in Plain, Glass Red Top tubes OR Plastic, Red Top Tubes, with Clot Activator; no other collection tubes are recommended for use, including gel serum separation tubes (SST). If plasma, the specimens MUST have originated from anticoagulated blood samples drawn in EDTA or Sodium Citrate collection tubes.
Storage ConditionsPIFA PLUSS™ PF4 assays must be stored refrigerated (2 to 8°C (36 to 46°F)).
Devices that have been removed from refrigeration, left in their sealed pouches, and stored at an ambient temperature (18 to 27°C; 64 to 81°F) for a maximum of 8 hours, may be returned to refrigeration for future use. This cycle can be repeated for a maximum room temperature exposure of 32 hours.
Please note:• Do not freeze tests.• If the test is frozen upon receipt, results will be invalid.• Do not use any tests beyond their expiration date.
Warnings and Precautions• For in vitro Diagnostic Use.• Do not expose the PIFA PLUSS™ PF4 MiniReactors to
temperatures greater than 40°C (104°F) or below 0°C (32°F).• Allow each PIFA® MiniReactor to warm to an ambient
temperature (18 to 27°C; 64 to 81°F), in the individual sealed pouch, for a minimum of 30 minutes prior to performing the test.
• PIFA PLUSS™ PF4 must be performed using anticoagulated samples draw with EDTA or Sodium Citrate blood collection tubes.
• All patient specimens should be collected using aseptic technique and should be handled in accordance with good laboratory practices.
• Frozen, thawed, hemolyzed, icteric, lipemic (of an excessive nature), bacterial contaminated specimens, or controls from other test kits should not be used, and can produce erroneous results.
• Inadequate incubation time, incomplete mixing, or improperly performed test procedures can produce erroneous results.
• Do not initiate testing with the PIFA PLUSS PF4 assay if the membrane extending from the accompanying seraSTAT™ is torn or creased.
• See also “Limitations.”
Limitations• The PIFA PLUSS™ PF4 should be used for the qualitative
detection of any antibody directed against the PF4 complex, and should be used as a screening test. There may be some antibodies reactive to the Heparin/PF4 complex that are non-reactive with this test. Test result should not be relied upon solely to identify an antibody to the PF4 complex.
• A positive test result may be indicative of a Heparin/PF4-related antibody in the test sample. However, the presence of these antibodies does not confirm the diagnosis of HIT or HITT. Therefore, results obtained from the PIFA PLUSS™ PF4 should be interpreted along with clinical findings and/or other serological tests.
Performance CharacteristicsAkers Biosciences, Inc. has conducted a series of evaluations to determine the performance of the PIFA® Heparin/PF4 Rapid Assay for the detection of antibodies to the Heparin/PF4 complex.
Studies were performed by outside laboratories to determine the performance of the PIFA® Heparin/PF4 Rapid Assay compared to standard laboratory methods using samples originating from field sources. The standard laboratory method was a commercially available ELISA technique.
ReproducibilityThe reproducibility of the PIFA® Heparin/PF4 Rapid Assay in detecting Heparin/PF4 antibodies was demonstrated by testing 10 aliquots of 5 specimens for inter-day evaluation and 10 aliquots for intra-day evaluation. Reproducibility of the PIFA® Heparin/PF4 Rapid Assay was determined to be 100% in both studies.
References1 Warkentin TE, Greinacher A, eds. Heparin-induced
thrombocytopenia, 3rd ed. New York: Marcel Dekker, Inc. 2004, 53, 205, 271, 295.
2 Daneschvar, H.Leon, MD, Daw, Hamed, MD: How to Interpret and Pursue an Abnormal Complete Blood Cell Count in Adults. Mayo Clinic Proc. 2005;80:1389-1394 2005 Mayo Foundation for Medical Education and Research.
3 CLSI (Clinical and Laboratory Standards Institute) H18-A4: Procedures for the Handling and Processing of Blood Specimens; Approved Guideline -- 4th ed. (ISBN 1-56238-72-3).
Specificity and Sensitivity ELISA
Positive Negative
PIFA®
Positive 21 3
Negative 2 153
• Specificity: 98.1% • Sensitivity: 91.3% • Overall Agreement: 97.2%
STEP 6 (continued):B. Tilt the MiniReactor 45° so the TOWER portion is elevated. Tap the Results Flange with finger until a BLUE/GREEN color appears to fill approximately 50% of the Reagent Window.
TEST PROCEDURE NOTE: The seraSTAT™ blood cell separator MUST be used to both extract a measured volume of the liquid fraction from the patient’s FRESH, Whole Blood specimen and dispense it into the PIFA PLUSS™ PF4 MiniReactor.
STEP 1: Ensure that the Whole Blood specimen is thoroughly mixed. Using a calibrated pipettor, dispense 150µL of the specimen onto the seraSTAT™ sample well.
Keep seraSTAT™ stationary until YELLOW reaches the LOWER EDGE of the membrane. Most samples will separate within five (5) minutes; occasionally separation may take up to eight (8) minutes.
STEP 3:Pick up the seraSTAT™ at the clear plastic sleeve. Rotate the seraSTAT™ so the YELLOW membrane is hanging in a downward position and the SAMPLE WELL is facing AWAY from you. Locate the seraSTAT™ above the TOWER, making sure that the YELLOW membrane can be inserted through the middle of the SLOT. Continue Inserting into the SLOT until the seraSTAT™ reaches a seated position.
STEP 4:With your flat palm(s), apply pressure to the seraSTAT™ and push DOWN until the seraSTAT™ and TOWER reach a FULL SEATED position. You will hear a “CRACK” as the reagents contained within the ampoule are released into the Reaction Chamber.
NOTE: the small but visible gap present when the Tower/seraSTAT™ have reached a FULL SEATED position.
STEP 5:Initiate 65-second timer. Vigorously, slide the device from side-to-side on the flat surface for 5 seconds (approximately 12 to 15 times). Stop motion when timer reaches “60”.
STEP 6:
A. Keep device stationary for the remaining 60 seconds.When timer sounds, IMMEDIATELY Pull the TOWER UP to the STOP position.
Akers Biosciences, Inc.
Please review the entire Package Insert and the labeled diagrams (Figures 1 & 2 BELOW) prior to initiating the PIFA PLUSS™ PF4 Assay Procedure
PRE-TEST PREPaRaTiONComplete in the following order:1. Remove the PIFA PLUSS™ PF4 sealed pouch from
refrigeration.
2. Visually inspect the pouch to confirm date of use is prior to expiration date.
3. The sealed pouch must remain at an ambient temperature (18 to 27ºC; 64 to 81ºF) for a minimum of thirty (30) minutes.
4. Open the pouch. Remove the PIFA PLUSS™ PF4 MiniReactor and place on a flat surface. Remove the seraSTAT™ Blood Cell Separator packet; open packet and place seraSTAT™ on a flat surface. Ensure that both components are NOT COOL to the touch. If they are COOL, allow to warm to an ambient temperature. Use an insulator between the units and the countertop if the latter is COOL.
5. Label the PIFA PLUSS™ PF4 MiniReactor with the patient’s identification. Ensure that the PIFA PLUSS™ PF4 logo on the device label is facing you.
6. Set timing device to 65 seconds; DO NOT initiate timing.
Thorofare, New Jersey 08086 USA For Technical Service, please call
1.800.451.8378 or +1.856.848.8698. Visit us on the web at www.akersbiosciences.com
STEP 2:Keep seraSTAT™ flat and gently slide clear, plastic sleeve over the sample well until the top edge of the sleeve is flush with the top edge of the seraSTAT™.
NOTE: YELLOW color may not be uniformly distributed throughout the membrane and may appeared mottled.
PIFA PLUSS™ PF4 MiniReactor
REaGENT WiNDOW
TOWER
SaMPLE WELL
LOWER EDGE
MEMBRaNE
CLEaR PLaSTiC SLEEVE
TOP EDGE
RESULT FLaNGE
CONTROL WiNDOW
TEST WiNDOW
SLOT
LOWER EDGE
TOP EDGE
PN 1300160 R2 10/12
STEP 7:
Return device to a flat surface. When a RED color appears in the CONTROL Window, interpret the result in the TEST Window in well-lit conditions.
RESULT iNTERPRETaTiONNEGaTiVE/Non-Reactive:TEST Window = BLUE-GREEN
NOTE: May contain various hues of BLUE-GREEN as the color is sample dependent.
POSiTiVE/Reactive:TEST Window = YELLOW
NOTE: • Flow rate is sample dependent. The time interval for
RED to develop in the CONTROL Window varies and ranges from 1-to-10 minutes.
• If RED fails to appear in the CONTROL Window, beyond the 10-minute mark, the TEST result is considered INVALID.
• Test Result is stable for thirty (30) minutes.
Figure 1
Figure 2
