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K1Z35 ?BIOPLEX 2200 VASCULITIS KIT, CALIBRATORS AND CONTROLS 510(k) SUMMARYThis summary of 510(k) safety and effectiveness information is being submitted in accordancewith the requirements of SMVDA 1990 and 21 CFR 807.92. 510(k) Numbber510(k) Summary Report DateOctober 24, 2007K072358MANUFACTURER INFORMATIONManufacturerBio-Rad Laboratories, Inc.Clinical Systems Division4000 Alfred Nobel DriveManufacturer AddressTelephoneEstablishment Registration No.Owner I OperatorHercules, CA 94547(510) 724-70002915274Bio-Rad Laboratories, Inc.4000 Alfred Nobel DriveHercules, CA 94547Owner I Operator No.9929003Official Correspondent for the BioPlex 2200 VasculitisOfficial Correspondent Address Bio-Rad Laboratories6565 185"'h Ave NETelephoneEstablishment Registration No.Owner I OperatorRedmond, WA 98052425-881-83003022521Bio-Rad Laboratories6565185"'h Ave NEOfficial CorrespondentTelephoneFaxRedmond, WA 98052Mr. Christopher Bentsen(425) 498-1709(425) 498-1651CLASSIFICATION INFORMATIONTest System, Antineutrophil Cytoplasmic Antibodies (ANCA),Devices, Measure, Antibodies to Glomerular BasementClassification NameCommon Name:Product Trade NameBioPlexDevice ClassClassification PanelRegulation NumberMembrane (GBM)Multi-Analyte Detection System VasculitisBioPlex 2200 Vasculitis kit on the BioPlex 2200 Multi-AnalyteDetection SystemBioPlex 2200 Vasculitis Calibrator Set2200 Vasculitis Control SetClass 11Immunology866.5660-Multiple Autoantibodies Immunological Test SystemBioPlex 2200 Vasculitis KitCalibrator Set and Control Set 510(k) Summary (10-24-07)Page 1 of 21
LEGALLY MARKETED EQUIVALENT (SE) DEVICESComparative FDA ClearedPREDICATE DEVICEPhadia Varelisa T M MPO EIAPhadia Varelisa T M PR3 EIAINOVA QUANTA Lite TM GBM ELISAINOVA NOVA Lite ANCA (Ethanol-fixed ate06/16/0407/02/0402/0819907/10/96DEVICE DESCRIPTIONThe Vasculitis kit uses multiplex flow immunoassay, a methodology that greatly resemblestraditional EIA, but permits simultaneous detection and identification of many antibodies in asingle tube. Three (3) different populations of beads are coated with antigens associated withvasculitis disease (MPO, PR3 and GBM).The BioPlex 2200 System combines an aliquot ofpatient sample, sample diluent, and bead reagent into a reaction vessel. The mixture is incubatedat 37 C. After a wash cycle, anti-human IgG antibody, conjugated to phycoerythrin (PE), is addedto the dyed beads and this mixture is incubated at 370C. The excess conjugate is removed inanother wash cycle, and the beads are re-suspended in wash buffer. The bead mixture thenpasses through the detector. The identity of the dyed beads is determined by the fluorescence ofthe dyes, and the amount of antibody captured by the antigen is determined by the fluorescenceof the attached PE. Raw data is calculated in relative fluorescence intensity (RFI).Three additional dyed beads, an Internal Standard Bead (ISB), a Serum Verification Bead (SVB)and a Reagent Blank Bead (RBB) are present in each reaction mixture to verify detectorresponse, the addition of serum or plasma to the reaction vessel and the absence of significantnon-specific binding in serum or plasma. Refer to the BioPlex 2200 System Operation Manual formore information.The instrument is calibrated using a set of four (4) distinct calibrator vials, supplied separately byBio-Rad Laboratories. A combination of four (4) vials representing four (4) different antibodyconcentrations are used for semi-quantitative calibration. The result for each of these antibodiesis expressed as an antibody index (Al).KIT COMPONENTSBioPlex 2200 Vasculitis reagent pack (Catalog No. 665-1850). The reagent pack containssupplies sufficient for 100 tests.VialDescriptionBead SetOne (1) 10 mL vial, containing dyed beads coated with Myeloperoxidase (MPO),Proteinase-3 (PR3) and Glomerular Basement Membrane (GBM); an InternalStandard bead (ISB), a Serum Verification bead (SVB), and a Reagent Blankbead (RBB) in a MOPS (3-[N-Morpholino] propanesulfonic acid) buffersupplemented with Glycerol and protein stabilizers (bovine). ProClin 300(0.3%), sodium benzoate (0.1%) and sodium azide ( 0.1%) are added aspreservatives.BioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 2 of 21
ConjugateFCartN-J-1One (1) 5 mL vial, containing phycoerythrin conjugated murine monoclonal anti-human IgG and phycoerythrin conjugated murine monoclonal anti-human FXIIIin phosphate buffer supplemented with murine and bovine protein stabilizers.ProClin 300 (0.3%), sodium benzoate (0.1%) and sodium azide ( 0.1%) areadded as preservatives.SampleDiluentOne (1) 10 mL vial, containing bovine and murine protein stabilizers intriethanolamine buffer. ProClin 300 (0.3%), sodium benzoate (0.1%) andsodium azide ( 0.1%) are added as preservatives.ADDITIONAL REQUIRED ITEMS, AVAILABLE FROM BIO-RADDescription663-1800BioPlex 2200 Vasculitis Calibrator Set: Four (4) 500 pL vials, each containinghuman antibodies to MPO, PR3 and GBM, in a human serum matrix made fromdefibrinated plasma. All calibrators contain ProClin 300 (0.3%), sodiumbenzoate (0.1%) and sodium azide ( 0.1%) as preservatives.663-1830BioPlex 2200 Vasculitis Control Set: Two (2) 1.5 mL Positive Control serum vials,each containing human antibodies to MPO, PR3 and GBM, in a human serummatrix made from defibrinated plasma; and two (2) 1.5 mL Negative Control serumvials, in a human serum matrix made from defibrinated plasma. All controlscontain ProClin 300 (0.3%), sodium benzoate (0.1%) and sodium azide ( 0.1%)as preservatives.660-0817BioPlex 2200 Sheath Fluid: Two (2) 4 L bottles containing Phosphate BufferedSaline (PBS). ProClin 300 (0.03%) and sodium azide ( 0.1%) are added aspreservatives.660-0818BioPlex 2200 Wash Solution: One (1) 10 L bottle containing Phosphate BufferedSaline (PBS) and Tween 20. ProClin 300 (0.03%) and sodium azide ( 0.1%)are added as preservatives.660-0000BioPlex 2200 Instrument and Software SystemBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 3 of 21
INTENDED USEBioPlex TM 2200 Vasculitis KitThe BioPlex TM 2200 Vasculitis kit is a multiplex flow immunoassay intended for the semiquantitative detection of IgG autoantibodies to Myeloperoxidase (MPO), Proteinase 3 (PR3) andGlomerular Basement Membrane (GBM) in human serum. In conjunction with clinical findings, thetest system is used as an aid in the diagnosis of anti-neutrophil cytoplasmic antibodies (ANCA)associated vasculitides: Microscopic Polyangiitis (MPA), Necrotising Glomerulonephritis, ChurgStrauss Syndrome, Wegener's Granulomatosis and the autoimmune renal disorder, Goodpasture'ssyndrome.The BioPlex 2200 Vasculitis kit is intended for use with the Bio-Rad BioPlex 2200 System.BioPlex 2200 Vasculitis Calibrator SetThe BioPlex 2200 Vasculitis Calibrator Set is intended for the calibration of the BioPlex 2200Vasculitis Reagent Pack.BioPlex 2200 Vasculitis Control SetThe BioPlex 2200 Vasculitis Control Set is intended for use as an assayed quality control tomonitor the overall performance of the BioPlex 2200 Instrument and BioPlex 2200 VasculitisReagent Pack in the clinical laboratory. The performance of the BioPlex 2200 Vasculitis ControlSet has not been established with any other Vasculitis assays.INDICATIONS FOR USEThe BioPlex 2200 Vasculitis kit is a multiplex flow immunoassay intended for the semiquantitative detection of IgG autoantibodies to Myeloperoxidase (MPO), serine proteinase 3(PR3) and Glomerular Basement Membrane (GBM) in human serum.The BioPlex 2200 Vasculitis kit is intended for use with the Bio-Rad BioPlex 2200 System.Uses:The test system is used to detect the presence of antibodies in serum samples, as an aid in thediagnosis of certain autoimmune vasculitides such as Microscopic Polyangiitis (MPA), NecrotisingGlomerulonephritis, Churg-Strauss Syndrome, Wegener's Granulomatosis and autoimmune renaldisorders, such as Goodpasture's syndrome, in conjunction with clinical findings and otherlaboratory tests.TECHNOLOGICAL CHARACTERISTICSThe following tables summarize similarities and differences between the BioPlex 2200 VasculitisKit and the predicate devices used in comparative studies with the BioPlex 2200 Vasculitis Kit.A. BioPlex 2200 Vasculitis vs. Predicate Phadia Varelisa MPO ANCA EIATable 1: Similarities betheen reagents and materialsSimilarities betweenBioPlex 2200 VasculitisComponents / Materials IReagentsWash Buffer, Sample DiluentPredicate Phadia Varelisa MPOANCA EIAWash Buffer, Serum DiluentBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 4 of 21
CalibratorsControlsjCalibratorsCalibratorsNegative Control, Positive Control[Negative Control and Multi-AnalytePositive Control (MPO, PR3 and GBM)Table 2: Similarities between reagents with regardto function and usePredicate Phadia Varelisa MPOBioPlex 2200 VasculitisSimilarities betweenANCA E1AFunction and Useand qualitativeSemi-quantitativeSemi-quantitative detection of IgGIntended Useautoantibodies to MPO, PR3 and GBM determination of MPO Anti neutrophilcytoplasmic antibodies in human serumin human serum.or plasma.SerumSerumMatricesTable 3: Differences between reagents and materialsBioPlex 2200 VasculitisDifferences betweenoomponents / MaterialsSolid PhaseReagentsSheath FluidPredicate Phadia Varelisa M POANCA EIA96 well microplate - antigen coatedmicrowellsConjugate: Anti-human IgG HRPconjugate/Substrate (TMB)PhycoerythrinSheath Fluid is used to suspend the bead Not similar; not utilized in EIA'sreagent and introduce it into the detector.Bead reagent - dyed antigen coatedbeadsConjugate: Anti-human lgG /Table 4: Differences between reagents with regard toifunction and usePredicate Phadia Varelisa MPOANCA EIADifferences betweenFunction and UseBioPlex 2200 VasculitisAnalyte DetectionSingle Analyte Detection (Human IgGMulti-Analyte Detection (human IgGautoantibodies to MPO, PR3 and GBM) antibodies to MPO)PlasmaSerumMatricesB. BioPlex 2200 Vasculitis vs. Predicate Phadia Varelisa PR3 ANCA EIATable 5: Similarities betveen reagents and materialsBioPlex 2200 VasculitisPredicate Phadia Varelisa PR3ReagentsWash Buffer, Sample DiluentANCA EIAWash Buffer, Sample gative Control, Positive ControlNegative Control and Multi-AnalytePositive Control (MPO, PR3 and GBM)Similarities betweenComponents / MaterialsBioPlex 2200 Vasculitis KitCalibrator Set and Control Set 510(k) Summary (10-24-07)Page 5 of 21
Table 6: Similarities between reagents with regardto function and usePredicate Phadia Varelisa PR3BioPlex 2200 VasculitisSimilarities between4NCA EIA/MaterialsComponentsSemi-quantitative and qualitativeSemi-quantitative detection of lgGIntended Useautoantibodies to MPO, PR3 and GBM etermination of PR3 Anti neutrophilytoplasmic antibodies in human serumin human serum.or plasma.SerumSerumMatricesTable 7: 1ifferences between reagentsand materialsDifferences betweenComponents / MaterialsSolid PhaseReagentsSheath FluidHioPlex 2200 VasculitisPredicate Phadia Varelisa PR3ANCA EIA96 well microplate - antigen coatedBead reagent - dyed antigen coatedmicrowellsbeadsC onjugate: Anti-Human IgG HRPConjugate: Anti-human igGonjugate /Substrate (TMB)PhycoerythrinSheath Fluid is used to suspend the bead ot similar; not utilized in EIA's.reagent and introduce it into the detector.Table 8: Differences between reagents with regardto function and use[Predicate Phadia Varelisa PR3BioPlex 2200 VasculitisDifferences betweenANCA EIAFunction and UseSingle Analyte Detection (Human IgGMulti-Analyte Detection (human lgGAnalyte Detectionautoantibodies to MPO, PR3 and GBM) antibodies to PR3)PlasmaSerumMatricesC. BioPlex 2200 Vasculitis vs. Predicate INOVA QUANTA-Lite GBM ELISATable 9: Similarities between reagents and materialsBioPlex 2200 VasculitisSimilarities betweenComponents / MaterialsWash Buffer, Sample DiluentReagentsControlsPredicate INOVA QUANTA-LiteGBM ELISAWash Buffer, Serum DiluentNegative Control, Positive ControlNegative Control and Multi-AnalytePositive Control (MPO, PR3 and GBM)Table 10: Similarities between reagents with regard tofiunction and usePredicate INOVA QUANTA-LiteBioPlex 2200 VasculitisSimilarities betweenGBM ELISAComponents / MaterialsSemi-quantitative determination of antiSemi-quantitative detection of IgGIntended Useautoantibodies to MPO, PR3 and GBM neutrophil cytoplasmic antibodies toGBM in human serumin human serum.7Table 11: Differences between reagents and materialsBiioPlex 2200 VasculitisDifferences betweenComponents / Materialsead reagent - dyed antigen coatedSolid PhasejbeadsPredicate INOVA QUANTA-LiteGBM ELISA96 well microplate - antigen coatedmicrowellsBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 6 of 21
ReagentsConjugate: Anti-human IgG /Phycoerythrinonjugate: Anti-Human lgG / Horse-Sheath Fluidradish Peroxidase, Substrate (TMB)Sheath Fluid is used to suspend the bead Not similar; not utilized in EIA's.reagent and introduce it into the detectorCalibratorsCalibratorsoneTable 12. Differences between reagentswith regardto function and useDifferences betweenBioPlex 2200 VasculitisPredicate INOVA QUANTA-LiteELISAGBMFunction and UseAnalyte DetectionMulti-Analyte Detection (human IgGSingle Analyte Detection (Human IgGautoantibodies to MPO, PR3 and GBM) antibodies to GBM)D. BioPlex 2200 Vasculitis vs. Predicate INOVA NOVA Lite, ANCA, Ethanol Fixed SlidesTable 13: Similarities between reagents and materialsPredicate INOVA NOVA Lite,BioPlex 2200 VasculitisSimilarities betweenComponents / Materials.NCA,Ethanol Fixed SlideslEA Negative Control, p-ANCANeegative Control and Multi-AnalyteControlsPositive Control (MPO, PR3 and GBM) Positive Control, c-ANCA PositiveJontrolTable 14: Similarities between reagents with regardto function andSimilarities betweenBioPlex 2200 VasculitisComponents / MaterialsANCA,Intended UseSemi-quantitative detection of lgGautoantibodies to MPO, PR3 and GBMin human serum.MatricesSerumusePredicate INOVA NOVA Lite,Ethanol Fixed SlidesSemi-quantitative determination ofAnti neutrophil cytoplasmic antibodiesin human serum.SerumTable 15: Differences between reagents and materialsDifferences betweenBioPlex 2200 VasculitisComponents / MaterialsBead reagent - dyed antigen coatedSolid PhasebeadsConjugate: Anti-human IgGReagentsSheath FluidCalibratorsPredicate INOVA NOVA Lite,ANCA, Ethanol Fixed SlidesANCA, Ethanol fixed neutrophilsSubstrate slidesC onjugate: Anti-Human IgG / HorsePhycoerythrinradish PeroxidaseSheath Fluid is used to suspend the bead Not similar; not utilized in EIA's.reagent and introduce it into the detector.CalibratorsNoneTable 16: Differences between reagents with regardto,function and useDifferences betweenIioPlex 2200 Vasculitis,Predicate INOVA NOVA Lite,Function and UsejNCA,Ethanol Fixed SlidesAnalyte DetectionMulti-Analyte Detection (human IgG}Single Analyte Detection (Human IgG}autoantibodies to MPO, PR3 and GBM) %antibodies)BioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 7 of 21
Solid PhaseBead reagent - dyed antigen coatedbeadsNCA, Ethanol fixed neutrophilsSubstrate slidesPERFORMANCE SUMMARYA. Expected ValuesExpected values for the BioPlex 2200 Vasculitis kit are presented in the following tables for serumsamples from normal blood donors (N 293) and unselected patient samples previously tested withvasculitis tests (N 300). A total of 300 serum samples from the normal blood donor population weretested. Seven (7) samples from the normal blood donor population were excluded due to "SerumVerification Bead (SVB) signal too low" analysis error messages during BioPlex 2200 Vasculitis kittesting. For all analytes, results of 1.0 Al are negative and results of 1.0 Al or greater are reportedas positive.Table A. BioPlex 2200 Vasculitis Kit Normal Blood Donors (N 293)BioPlex ResultPositiveNegativeAnti-MPO01293 (0.0%)293293 (100.0%)Anti-PR30/293 (0.0%)293/293 (100.0%)Anti-GBM2/293 (0.7%)291/293 (99.3%)Table B. BioPlex 2200 Vasculitis Kit Unselected Patient Samples PreviouslyTested With Vasculitis Tests (N 300)PositiveNegativeAnti-MPO14/300 (4.7%)286/300 (95.3%)Anti-PR381300 (2.7%)292/300 (97.3%)Anti-GBM1/300 (0.3%)299/300 (99.7%)BioPlex ResultB. Reproducibility StudiesA reproducibility panel, consisting of ten (10) serum panel members, was prepared by Bio-RadLaboratories. The positive panel members were prepared by combining patient samples positive forBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 8 of 21
antibodies to MPO, PR3 and GBM. Two (2)of the ten (10) had high levels of the antibodies toMPO, PR3 and GBM; two (2) members had low levels of the antibodies to MPO, PR3 and GBM;and two (2) members had antibody levels near the cutoff. There was also one (1) high negative andone (1)low negative panel member. In addition, a BioPlex 2200 Vasculitis positive control (antibodypositive for MPO, PR3 and GBM) and a negative control (antibody negative for all 3 analytes) wereincluded and tested as panel members.Reproducibility testing was performed at two (2)US testing facilities and an internal site (Bio-RadLaboratories) on a total of two (2) lots of the BioPlex 2200 Vasculitis kit. The ten (10) panelmembers were provided to the each of the testing sites. Two (2) of the three (3) testing facilitiesevaluated reproducibility using one (1) kit lot of the BioPlex 2200 Vasculitis kit and the third siteevaluated the second lot of the BioPlex 2200 Vasculitis kit. Each of the ten (10) panel members wastested in duplicate (x2) on two runs per day for three days at each testing site (2 times x 2 runs x 3days x 3 sites 36 replicates per panel member and controls). The data were then analyzed forintra assay and inter-assay reproducibility according to the Clinical and Laboratory StandardsInstitute guidance (formerly NCCLS) EP5-A2, revised November 2004 and ISO/TR 22971:2005.The mean Antibody Index (Al), standard deviation (SD), and percent coefficient of variation (%CV)for each panel member were calculated. Results can be found in the below table:Table. Reproducibility; BioPlex 2200 VasculitisBioPlex 2200 Vasculitis KitVasculitisKit 00.2777.0%0.156365.50.2955.4%0.249 4.5%0.000 0.0171.2%0.089 1.3% 4.6%7.9%1HighPositiveo8.4%2 ,c LowPositive1LowBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 9 of 210.082
Cutoff 1a·E. NearCutoff 1%0.0000.0%0.0855.5%0.15710,21NearCutoff %NearCutoff .0%0.02712.6%2410.9%00410 BioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 10 of 21188
%NearCutoff NearCutoff 0%0.00000%0.00000%0 * Between-site0.0410%000variance includes between lot varianceC. Comparative TestingThree hundred (300) normal blood donors and three hundred (300) unselected patient samplespreviously tested with vasculitis tests were tested with the BioPlex 2200 Vasculitis kit. Seven (7) ofthe three hundred (300) normal blood donor samples were excluded due to "Serum VerificationBead (SVB) signal too low" analysis error during BioPlex 2200 Vasculitis kit testing. All sampleswere also tested by the corresponding commercially available microplate EIA methods. The resultscan be observed in Tables A - F.BioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 11 of 21
Table A. BioPlex 2200 vs. Anti-MPO EIA - Normal Blood Donors (N 293)BioPlex 2200 Vasculitis Anti-MPO Result ·MPositive'-. , EquivocalEquivocalEE ,' m·000'o0.Z000000000100.0%, toNegative0293293Total0293293MM 00000- 0o- E0 --N/A N/A(293/293)98.7%,100%100.0%(293/293)98.7%,100%N/A Not ApplicableTable B. BioPlex 2200 vs. Anti-PR3 EIA - Normal Blood Donors (N 293)BioPlex 2200 Vasculitis Anti-PR3 Result o.a.i00i;Z0 293Cio-toAN/A4.U E 0-0E o-100.0%(293/293)98.7%,N/A Not ApplicableBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 12 of 21100.0%100%(293/293)98.7%,100%
SITable C. BioPlex 2200 vs. Anti-GBM EIA - Normal Blood Donors (N 293)BioPlex 2200 Vasculitis Anti-GBM Result04zIL 2NegativePositive02*Negative2289291Total229129329 'E- --Z82 E E .12--N/AN/AN/AN/A (289/291)98.6% 99.3%97.5%,99.8%(289/293)96.5%,99.5%* Two (2) positive anti-GBM EIA results were weak positive.N/A Not ApplicableTable D. BioPlex 2200 vs. Anti-MPO EIA - Unselected Patient Samples PreviouslyTested With Vasculitis Tests (N-300)BioPlex 2200 Vasculitis Anti-MPO Result-I tal14286300---71.4%(5/7)35.9%,91.8%97.6%(284/291)0 -0a--a--95.1%,98.8%Two (2) anti-MPO EIA equivocal results are included in the Overall Agreement.BioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 13 of 2196.3%(289/300)93.6%,97.9%
Table E. BioPlex 2200 vs. Anti-PR3 EIA - Unselected Patient Samples PreviouslyTested With Vasculitis Tests (N 300)BioPlex 2200 Vasculitis Anti-PR3 Result I-o L .Equivocal'5 0 0w[o[o--'7 o.2zo)55 OPositive--.Q Z0 100.0%(5/5) .1%,99,7%99.0%(297/300)97%,99.7%295 300o. - 4). 4 2r0 sf If.4Table F. BioPlex 2200 vs. Anti-GBM EIA - Unselected Patient Samples PreviouslyTested With Vasculitis Tests (N 300)BioPlex 2200 Vasculitis Anti-GBM Result 5Positive,.,ega vo {Total0100.0%298299 8 299 ---1N ot --0ZI- (5/55%Not 99.7%100Accurate% (298/299) 0/1)98.1%,99.9%99.3%(298/300)97.6%,99.8%300· One (1) anti-GBM EIA results was a weak positive.kit was further evaluated by testing 227 retrospective samples positiveVasculitis2200The BioPlexfor anti-MPO (N 1 00), anti-PR3 (N 1 00), and anti-GBM (N 27). All samples were also tested bythe corresponding commercially available microplate EIA methods. In addition, the anti-MPO andanti-PR3samplscuritested by an ANCA IFA method using ethanol-fixed slides) ThepositiveBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 14 of 21
results can be observed in Tables G - K.Table G. BioPlex 2200 vs. Anti-MPO EIA - Retrospective Anti-MPO Positive Samples (N 100)BioPlex 2200 Vasculitis Anti-MPO Result0M0UM 93.9%87.3%,:5(92/98) oNegativeTotal930117100 e (1) anti-MPO EIA equivocal result is included in the OverallAgreement. Not ApplicableN/ATableH. BioPlex 2200 vs. Anti-PR3 EIA - Retrospective Anti-PR3 Positive Samples(N 100)BiaPlex 2200 Vasculitis Anti-PR3 Result-o ')40 4(0a- AN/A83.0%(83/100)LUI*Ten (10) anti-PR3 EIA equivocal results are included in the Overall Agreement.*N/A Not ApplicableSioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 15 of 2174.4%,89.1%
Table I. BioPlex 2200 vs. Anti-GBM EIA - Retrospective Anti-GBM Positive Samples(N 27)BioPlex 2200 Vasculitis Anti-GBM gativ02**- 110 0E, , Oc18- · -- E . .·· 9%27*Two (2) of the sixteen (16) anti-GBM F/A positive results were weak positive.positive results wereweak positive.·* One (2) of the two (2) anti-GBM EIAThe remaining fifteen (15) of the eighteen (18) anti-GBM EIA results were moderate to strongpositive.N/A Not ApplicableTable J. BioPlex 2200 vs. pANCA IFA - Retrospective Anti-MPO Positive Samples(N 100)BioPlex 2200 Vasculitis Anti-MPO Result4' IL Z4)Positive83n.Negative10111Total937100-6o, -4o:· 0o- 89-86.1%,93.3%93.3%(83/89)969% N/AN/A -ZoN/A Not ApplicableBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 16 of 2184.0%(84/100)75.6%,899%
Table K. BioPlex 2200 vs. cANCA IFA - Retrospective Anti-PR3 Positive Samples(N 100)BioPlex 2200 Vasculitis Anti-PR3 ResultCz 0Cr00zPositive93598 %:394.9%(93/98)88.6%,97.8%N/AN/AU-4ZC)N/A Not ApplicableTables L and M compare EIA results from the retrospective positive samples for anti-MPO EIA(N 100) and anti-PR3 EIA (N 100) with ANCA IFA results.Table L. pANCA IFA vs. Anti-MPO EIA - Retrospective Anti-MPO Positive Samples(N 100)Anti-MPO EIA Result4 C0U's002 00) z 0 .,.,u9:2.Positive87,v-eatvTotal11-Negative1 0 10(874899810o00n, oCo00Q0.0'*-oz00o89-1197.8%(837/89)92.2%-N/A99.4%1 c.0 1004 · One (1) anti-MPO EIA equivocal result is included in the Overall Agreement.N/A Not ApplicableBioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 17 of 21N/A87.0%79%,(87/1 00)92.2%
Table M. cANCA IFA vs. Anti-PR3 EIA - Retrospective Anti-PR3 Positive Samples(N 100)Anti-PR3 EIA ResultO--) Co-Positive781010 [Negative1011ZTotal7910110)Eo0.ru4 ,i i -c- EECo98-21 79.6%(78/98)70.6%,86.4N/AN/A79.0%(79/100)100Ten (10) anti-PR3 EIA equivocal results are included inthe Overall Agreement.N/A Not ApplicableA combined positive agreement of 94.1% (176/187) was observed between BioPlex 2200 Vasculitisanti-MPO/anti-PR3 and ANCA IFA results (from Tables L and M), compared to a combined positiveagreement of 88.2% (165/187) between anti-MPO/anti-PR3 EIA and ANCA IFA results (fromTables N and 0). Also, a combined overall agreement of 88.5% (177/200) was observed betweenBioPlex 2200 Vasculitis anti-MPO/anti-PR3 and ANCA IFA results compared to a combined overallagreement of 83.0% (166/200) observed between anti-MPO/anti-PR3 and ANCA IFA results.Table N summarizes the test results (positive percent agreement, negative percent agreement andoverall percent agreement) for each of the three antibodies (anti-MPO, anti-PR3 and anti-GBM)from normal blood donors (N 293), unselected patient samples previously tested with vasculitistests (N 300), and retrospective positive samples (N 100 for anti-MPO, N 100 for anti-PR3 andN 27 for anti-GBM). These sample populations were tested by the BioPlex 2200 Vasculitis kit andanti-MPO, anti-PR3 and anti-GBM EIAs. In addition, anti-MPO and anti-PR3 known positivesamples were tested by an ANCA IFA method.BioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 18 of 2170.0%85.8%
Table N. Summary of Positive, Negative and Overall Percent Agreement for NormalBlood Donors (N 293), Unselected Patient Samples Previously Tested With VasculitisTests (N 300), and Retrospective Positive Samples (N 100 for anti-MPO, N 100 foranti-PR3 and N 27 for anti-GBM)Anti-MPO EIAResult EEC,Normal BloodDonors* N/AN3UnselectedNo 5/7Palient71.4% Samples(01pANCA IFAResult-EE 0))293/293 /10093 0%Anti-PR3 EIAResultEEE220' cANCA IFAResultEE)CNTNT5NTNT-E0'0'050 28 TN5/5292/29 295292/295 297/300NT10% 99099.0%NTNT799100 0%N/A20 NTAnti-GBM EIAResultN/A293/293 293/293100.0% 100.0%20 NEE-0628 2 28 /289/291 289/293993% 986%Not 298/299 298/300Accuat997% 993% m PreviotoslVPositive92/98939%83/81003NN/A 84 al blood donors and unselectedpatientsamplespreviously tested with vasculitis tests werenot tested by ANCA IFA.N/A Not ApplicableNT Not Tested*E. Cross-ReactivityA cross-reactivity study was performed to determine if samples from various disease states andother potentially interfering factors interfere with test results when tested with the BioPlex 2200Vasculitis kit. A panel of ten (10) specimens positive for each cross reactant were evaluated forpossible cross reactivity with the BioPlex 2200 Vasculitis kit for each of the three (3) autoantibodies.Samples were also tested on a corresponding commercially available microplate EIAs. Most of thesamples evaluated were high positive for each disease state. The results demonstrated that thevarious disease state samples evaluated do not cross react with the three (3) autoantibodies in theBioPlex 2200 Vasculitis kit. Results can be found in the below table:Table. Anti-PR3AntiGBMBioPlex 2200000EIA000Discrepants000BioPlex 2200 Vasculitis Kit,Calibrator Set and Control Set 510(k) Summary (10-24-07)Page 19 of 21
nti-dsDNAAnti-Histone10101010BioPlex 2200010EA010Discrepants000BioPlex 2200000EIA100Discrepants100BioPlex 2200000EIA0Discrepants000BioPlex 2200220EIA100Discrepants120BioPlex 22000Discrepants000BioPlex 2200000EIA000Discrepants000BioPlex 22000000000RheumatoidFactor (RF)Anti-ThyroidPeroxidase(T
The BioPlex 2200 Vasculitis kit is a multiplex flow immunoassay intended for the semi-quantitative detection of IgG autoantibodies to Myeloperoxidase (MPO), serine proteinase 3 (PR3) and Glomerular Basement Membrane (GBM) in human serum. The BioPlex 2200 Vasculitis kit is intended for use with the Bio-Rad BioPlex 2200 System.
