Heparin / PF4 Assayed Antibody Serum Panel Frequently Asked Questions

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1 Please Consult the Product Package Insert Prior to Use Intended Use Q. What is the Intended Use of the Heparin/PF4 Antibody Panel? A. The Heparin/Platelet Factor 4 Antibody Serum Panel is an assayed control, intended for use as a serum QC control to monitor and evaluate precision and accuracy of the (qualitative) PIFA Heparin/PF4 Rapid Assay. Included are both confirmed positive and negative control panel members. Q. What is the difference between the Assayed (PN & ) and Unassayed (PN & ) Heparin/PF4 Antibody Panels? A. The Heparin/PF4 Assayed Antibody Control Panels have been developed specifically for use with the PIFA Heparin/PF4 Rapid Assay. In addition to antibody classification, it also supplies HPF4 antibody analyte values for the control materials. The unassayed Heparin/PF4 Antibody Control Panels supply antibody information (positive/negative) but do not have manufacturer-assigned analyte values. These controls have been developed specifically for use with the PIFA PlussPF4 Rapid Assay and other qualitative Heparin/Platelet Factor 4 antibody detection systems that permit the use of serum-based materials for routine quality control. Storage and Reagent Handling Q. How should the Heparin/PF4 Antibody Serum Panel be stored? A. As noted in the Package Insert, serum panel members should be stored at -70 C or colder. These conditions are required to maintain antibody stability and achieve the labeled expiration dating. If serum panel members and/or aliquots are stored above -70 C, the samples may not perform per classification and can only be used for informational purposes not quality control. The labeled expiration dating would also be invalidated. Q. I don t have a -70 C freezer, do you have any suggestions for meeting my Quality Control needs for the PIFA Heparin/PF4 Rapid Assay? A. First, if possible, borrow space from another department who may have a -70 C or colder freezer. The control material is very small and doesn t take up much space. Controls should be assayed as required by your laboratory s standard quality control policy. As the manufacturer, Akers Biosciences recommends that the Laboratory run controls upon receipt of a new lot of PIFA Heparin/PF4 Rapid Assay devices and after Compiled by AKERS BIO, 201 Grove Road, Thorofare, New Jersey USA Page 1 of 6

2 running 100 devices within the same lot. As unit-use devices, Quality Control samples can be run less frequently. Some customers have found the following approach helpful. Since your distributor may not provide lot-specific information when you place your order for PIFA devices, when it is time to reorder, also order a new serum panel. Run controls upon receipt of your shipment, even if there are no patient samples in queue. This should keep you in compliance until the next shipment (depending on the QC policy of your laboratory). Q. Do I have to use a quick thaw procedure? A. Yes, proteins will precipitate out of frozen specimens if they are thawed at room temperature. The quick thaw procedure should minimize protein shedding. Our serum panel is frozen and comes on dry ice but it is processed under strict specifications in order to avoid the formation of particulates. Particulates could potentially interfere with the PIFA Heparin/PF4 Rapid Assay device performance. Q. If I don t have a water bath capable of maintaining C, can I use a heating block? A. It is generally not recommended to use a heat block. AKERS BIO has found there is a great deal of temperature variation when using a heat block. It is possible to overheat the sample and thus cause damage to the HPF4 antibodies. If no other method is available, AKERS BIO recommends you monitor the heat block while thawing the control samples. Place a small amount of liquid in a tube in the heat block. Place a calibrated thermometer into the tube and monitor to insure the block remains within C for the 6 to 7 minute thaw period. Q. What happens if I forget about the control and it is heated for an extended period of time? A. More than likely, the positive sample will not perform per classification. The sample in each cryovials can quickly be overheated. The proteins contained in the HPF4 positive sample will degrade if exposed to excessive heat and they will not activate the reagent in the PIFA device. The negative control will still work per classification but it is not a true indicator of device performance due to protein degradation. Q. The cryovial contains such a small volume, how can I accurately mix it? A. Inversion is not the mixing method of choice! We suggest the vial be vortexed to ensure mixing before use. It is important to have a homogenous dispersal of antibody throughout the sample. Insure sample is well mixed before use and before additional handling such as preparation of aliquots. Compiled by AKERS BIO, 201 Grove Road, Thorofare, New Jersey USA Page 2 of 6

3 Q. After I thaw the sample, how soon should I use it? A. It is best to use the sample immediately after thaw to maximize antibody reactivity. It is possible to keep the thawed sample at 2 to 8 C for a maximum of eight (8) hours prior to use. Remember: controls that have been kept refrigerated should warm to room temperature prior to use. This should take approximately thirty (30) minutes. Always mix well prior to use. Q. After use, how do I handle the remaining sample? A. When using HPF4 serum panel members from AKERS BIO, it is imperative to avoid multiple freeze-thaw cycles. Once thawed, the well mixed remainder of the control should be flash frozen, stored at 70 C and should only be subjected to one additional freeze-thaw cycle. When you are done using the initial sample, separate the remaining volume into 40µL single use aliquots and return these back into frozen conditions ( 70 C ). At a minimum, each panel member has 150µL starting volume. We recommend using the smallest possible cryovials for storage. If a 40µL aliquot is stored in a large volume tube, it is difficult to recover volume as the sample often coats the sides of the tube. It is also more prone to freezer burn due to the increased surface area. You should get one initial run and at least three (3) aliquots from each control vial. Pipetting technique will affect the yield obtained from each vial. Q. What is meant by Follow asceptic techniques during the aliquot procedure? A. To protect the sample aliquot from bacterial contamination follow standard asceptic techniques. This would include use of sterile pipet tips and cryovials. A bacterially contaminated sample may non-specifically block the membrane filtration system leading to false positives. Q. I don t have time to aliquot; can t I just freeze/thaw the same sample? A. No! Repeated freeze/thaw cycles may diminish antibody activity. The Heparin/PF4 Antibody Serum Panel members should only be exposed to two (2) freeze/thaw cycles; the initial thaw and one additional freeze/thaw cycle. AKERS BIO has validated the performance of the control under these conditions. Results Interpretation Q. How can I get an individual panel member s Certificate of Quality (C of Q) data? Compiled by AKERS BIO, 201 Grove Road, Thorofare, New Jersey USA Page 3 of 6

4 A. The antibody classification information (positive/negative) is provided on each control vial. Detailed C of Q data for each panel member can be found by looking up the Heparin/PF4 Antibody Serum Panel kit lot number (found on the outside of the kit box). This data can be accessed by visiting the Akers Biosciences Inc. website ( and going to the appropriate product webpage. Data is provided for informational purposes and is listed by Lot Number. If you wish to be mailed or faxed a copy of this information, contact technical service at (US: TEST). Q. Can I use a control from another Heparin/PF4 test kit with the PIFA Heparin/PF4 Rapid Assay devices? A. No! Most controls are plasma based. Plasma or converted plasma is not suitable to test with this assay and should not be used. Each control panel member in the HPF4 Assayed Antibody Serum Panel originates from a single donor serum unit. These antibody controls are manufactured under strict guidelines to give our customers the highest quality controls for use as validation and QC samples. When using controls from other test kits, AKERS BIO cannot guarantee their sample matrix would be compatible with this assay. Controls from other test kits should not be used. Q. What happens if my Heparin/PF4 antibody proficiency testing sample is plasma? A. It cannot be run on the PIFA Heparin/PF4 Rapid Assay devices per manufacturer specifications. Most likely, it will give an inaccurate result. Q. What can I do in the above circumstance? A. If this happens, consult your survey and the governing body to clarify their requirements. It may be possible to blind and run assayed Heparin/PF4 antibody panel members or an appropriately gathered characterized patient sample. Additionally, AKERS BIO is happy to announce that the College of American Pathologists (CAP) currently offers a proficiency panel for use specifically with the PIFA Heparin/PF4 Rapid Assay. The Coagulation Special Testing Series offers a Heparin-Induced Thrombocytopenia Module that has a program specific for AKERS BIO PIFA Heparin/PF4 Rapid Assay. Q. If the HPF4 serum panel members are frozen, why can't I use my frozen specimens? A. If not handled meticulously, freezing and thawing will decrease antibody activity, and could cause a positive sample to produce a negative test result. Freezing/thawing may also cause certain proteins to precipitate out of solution, and cause other microparticulates or debris to form. These particulates can clog the pores in the membrane filter system in the test device, and could cause a negative sample to produce a Compiled by AKERS BIO, 201 Grove Road, Thorofare, New Jersey USA Page 4 of 6

5 positive test result. AKERS BIO manufactures our serum panel members under very strict conditions to avoid the formation of any particulates and interfering substances. Q. I m using such a small volume of control (30µL), does the temperature really matter? A. Yes! The overall temperature of both the control and the devices is very important. They should both warm to room temperature for a minimum of 30 minutes prior to use. The colder the reagent/sample mixture, the more slowly the antibody/antigen reaction occurs. Q. I m just a little short in the volume of control I have, can I run it anyway? A. No, an incorrect amount of sample can affect the test result! Individual Quality Control Plan Q. Are the Assayed HPF4 Antibody Serum Panels used with a CLIA waived assay? A. No, the Assayed HPF4 Antibody Serum Panels are used with the PIFA Heparin/PF4 Rapid Assay which is classified as moderately complex by CLIA. Q. On December 31, 2015 the ability to use Equivalent Quality Control (EQC) policies expired. Laboratories are required to either follow the quality control regulations as outlined in CLIA 88 or develop an Individualized Quality Control Plan (IQCP) to replace the current EQC option. Does AKERS BIO supply manufacturer s recommendations to help meet the new quality control requirements under CLIA? A. Yes, Akers Biosciences Inc. has developed an IQCP Support Document, a useful template that can be customized for your laboratory s particular needs. It was developed using the Center for Medicare & Medicaid Services (CMS) and CDC publication Developing an IQCP, A Step By Step Guide. It is accompanied by a Training Quiz that can be used as part of your laboratory s ongoing training. Q. Does the template cover all aspects of the new CLIA requirements? A. No, although Akers Biosciences has provided a useful template for your use with our products, the CMS expects your IQCP to reflect your lab conditions and not what Akers Biosciences assumes about your laboratory operation and conditions. Q. Does my lab have to develop an IQCP for use with this product? A. No, Your laboratory director is responsible for deciding whether the laboratory will utilize IQCP and for ensuring that the quality control plan (QCP) developed effectively Compiled by AKERS BIO, 201 Grove Road, Thorofare, New Jersey USA Page 5 of 6

6 meets the IQCP requirements. Alternately, your lab can follow the Quality Control Regulations outlined in CLIA 88 to replace Equivalent Quality Control policies. Q. How can I get the IQCP Support Document & Training Quiz? A. Both the IQCP Support Document and the Training Quiz can be found and be downloaded from the Akers Biosciences website: or by calling TEST and requesting them. Compiled by AKERS BIO, 201 Grove Road, Thorofare, New Jersey USA Page 6 of 6