ARCHITECT cSystems OEM Reagent Training MULTIGENT Gentamicin NOTE: “Multigent” and “ARCHITECT cSystems” are trademarks of Abbott Laboratories in various jurisdictions. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Assay Gentamicin (Seradyn) Available on ARCHITECT cSystems only Launch Date – September 2006 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Reagent labeling will indicate: Manufactured by Seradyn Distributed by Abbott Laboratories Disk is currently available for c8000 and c16000. Disk for c4000 is targeted for 4Q 2009. No application for AEROSET. Abbott List Number on labeling. Abbott Laboratories Customer Support phone number on package insert. Reagent cross contamination, high sample carryover and required flagging were evaluated. Assay support is provided by Abbott. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Clinical Utility Gentamicin is used in the treatment of serious infections involving aminoglycoside-sensitive organisms (aerobic gram-negative bacteria). It has a narrow range of safe and effective concentration. Elimination of the drug, or clearance, is highly dependent on renal function, any impairment of glomerular filtration causes accumulation of the drug. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Reagent Methodology - Homogeneous particle-enhanced turbidimetric inhibition immunoassay (PETINIA) CAP Code – 1322 List Number 1E11-20 Reagent - 2 x 100 Test kit R1 = 2 x 25 mL (ready to use) R2 = 2 x 9 mL (ready to use) Storage 2-8°C Kit is stable to the expiration date On board stability is 55 days Do not Freeze or expose to temperatures above 32°C. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Reaction Mechanism Gentamicin + Antibody to Gentamicin /Trizma buffer (R1) + Gentamicin Coated Microparticles (R2) Competition Gentamicin: Antibody complex + Gentamicin-coated Microparticle: Antibody complex 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

Reaction Graphic Not for Customer Distribution     30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Calibrators List Number – 1E11-01 6 levels – 1 x 1 mL - Liquid ready to use Cal 1 – 0.0 mg/mL (0.00 mmol/L) Cal 2 – 0.5 mg/mL (1.05 mmol/L) Cal 3 – 1.5 mg/mL (3.14 mmol/L) Cal 4 – 3.0 mg/mL (6.27 mmol/L) Cal 5 – 6.0 mg/mL (12.54 mmol/L) Cal 6 – 10.0 mg/mL (20.90 mmol/L) 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Calibrators Cal curve stability is 28 days. Cal 1 (Zero Cal) is the calibration blank. Calibrator stability is indicated by the expiration date on the carton. Store tightly capped at 2 to 8°C. Do not freeze or expose to temperatures above 32°C. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin c8000 Calibration Curve Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Specimen Collection and Handling Serum or Plasma are acceptable specimens Serum — Glass or plastic tubes with or without a gel barrier and with or without clot activators. Note: Some gel separation tubes may not be suitable for use with TDM assays. Refer to the tube manufacturer’s labeling. Plasma — Lithium Heparin, Sodium Heparin, K2-EDTA, and K3-EDTA in glass or silicone-coated plastic tubes with or without a gel barrier. B-D gel barrier tubes equivalent to serum 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Specimen Storage Separated Sample Storage: 7 days at 2-8°C 14 days at less than -10°C Samples containing Carbenicillin or Piperacillin: These samples should be stored frozen if a delay in analysis is going to be more than 8 hours. Failure to freeze samples containing these antibiotics may result in falsely low gentamicin levels due to in vitro inactivation. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Performance Characteristics Acceptable Criteria is within 10% or 0.1 mg/mL Linearity 0.5 – 10.0 mg/mL (1.05 – 20.90 mmol/L) 10.0 mg/mL (20.90 mmol/L) is the highest level calibrator value Accuracy by Recovery Recovered concentrations ranged from 1.09 to 8.23 mg/mL (2.28 to 17.20 mmol/L) Recovery at the 4 levels ranged from 102.22 to 109.00 % 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Performance Characteristics, con’d Precision Acceptable Criteria is: <10% total CV for medium and high controls. < 7% total CV for low controls. Representative data for total run precision is shown below: Level 1 Level 2 Level 3 Mean (ug/mL) 2.45 6.10 9.12 Total SD 0.095 0.222 0.251 Total %CV 3.89 3.63 2.75 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Performance Characteristics, con’d Method Comparison ARCHITECT vs. FPIA ___________________________________ N 63 Slope 1.102 y-Intercept - 0.412 Correlation Coefficient 0.997 Range (mg/mL) 0.7 to 9.0 ___________________________________ 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Reagent Cross Contamination SmartWash information is included in the ASSAY PARAMETERS. R1/R2 Detergent A wash against “All” reagents R1/R2 Detergent A wash against “GENT9” Detergent A Sample Probe Wash The following cuvette washes: Albumin BCP – 0.5% acid wash AST - 0.5% acid wash ALT - 0.5% acid wash Trig - 10% Detergent B See the Gentamicin package insert, LIMITATIONS OF THE PROCEDURE, for assays that are affected by the Gentamicin reagent and require the addition of a GENT9 SmartWash. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

SmartWash configuration is critical… Gentamicin is used as a microbial agent, or preservative, in controls and some reagents (Albumin BCP, AST and ALT). Testing confirms sample probe wash is necessary to reduce carryover from high patient samples, but also from control samples containing gentamicin. Reagent washes are configured for R1/R2 for “All” reagents AND for the “GENT9” assay. Cuvette wash configuration is critical. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Interfering Substances Interference results for bilirubin, HAMA, hemolysis, RF, Total Protein, and lipids are listed in the package insert. Sisomicin cross-reacts with the gentamicin assay due to structural similarity (falsely elevates results). This assay can’t be used on samples from patients who are also taking sisomicin. See package insert for cross reactivity due to other drugs and Gentamicin metabolites. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

MULTIGENT Gentamicin Therapeutic and Toxic Ranges Peak (mg/mL) Trough (mg/mL) Traditional Dosing (Toxic Levels) >10 to 12 >2 to 4 Less Severe Infections 5 to 8 <1 Severe Infections Infections 8 to 10 <2 to 4 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

Auto-Dilution The systems are capable of auto-diluting using a 1:2 dilution with saline as the diluent. Auto-dilution is implemented in the assay parameters of package insert commodity 0148346B. 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

Questions and Answers Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2

Why is there no AEROSET application? Due to the Gentamicin reagent being very “sticky” and requiring special washes to reduce carry over, it was not possible to apply this assay to the AEROSET system. The ARCHITECT cSystem has better washing capabilities and improved wash materials that reduce the risk of carry over. End of Document 30-5394/R2 Not for Customer Distribution Located electronically in the c8000 Project Notebook 3.6.4 version 2