SIEMENS ADVIA Centaur®CP Immunoassay System Anti-HBe2 (aHBe2) Assay for the Detection of Antibodies to Hepatitis B e Antigen Current Revision and Date a Rev. A, DRAFT Product Name ADVIA Centaur® Anti-HBe2 (aHBe2) !REF! 10720831 Abbreviated Product Name ADVIA Centaur aHBe2 Test Name/ID aHBe2 Systems ADVIA Centaur CP system Materials Required but Not ADVIA Centaur aHBe2 Quality Control IREFI 10720832 Provided ADVIA Centaur Wash 1 (2 x 1500 mL) IREFI 01137199 ADVIA Centaur Wash 1 (2 x 2500 mL) IREFI 03773025 Specimen Types Serum, EDTA plasma, and lithium heparin plasma Sample Volume 75 µL Measuring Interval 0.00 –3.50 Index a A vertical bar in the page margin indicates technical content that differs from the previous version. Intended Use The ADVIA Centaur® Anti-HBe2 (aHBe2) assay is an in vitro diagnostic immunoassay for the qualitative detection of antibodies to the e antigen of the hepatitis B virus (HBV) in human pediatric (2-21 years old) and adult serum, EDTA plasma, or lithium heparin plasma using the ADVIA Centaur CP system. Assay results, in conjunction with other laboratory results and clinical information may be used as an aid in the diagnosis of hepatitis B virus (HBV) infection in patients with signs or symptoms of hepatitis B infection, or with risk factors for HBV infection, or with known HBV infection. Results of the assay, in conjunction with other diagnostic information, may be used to aid in determining HBV seroconversion. This assay is not intended for screening donors of blood or blood products or human cells, tissues, and cellular and tissue-based products (HCT/Ps). 10720831_CP_US_EN Rev. A, DRAFT 1 / 27 aHBe2 ADVIA Centaur CP System Summary and Explanation Hepatitis B is a potentially life-threatening liver infection caused by the hepatitis B virus (HBV). It is a major global health problem. It can cause chronic infection, which puts people at high risk of death from cirrhosis and liver cancer.1 Infection is transmitted from person to person by contact with infected body fluids. Sexual contact is an important mode of transmission, but infection is also transmitted by transfusion of contaminated blood or blood products, or by use of contaminated needles or syringes for injections. Perinatal transmission may occur from mother to baby.2 The average incubation period for HBV infection is 6–8 weeks (range 1–6 months). Common clinical symptoms include malaise, fever, gastroenteritis, and icterus. In adults, 90% to 95% of patients with HBV infection completely recover from acute illness and clear the virus. Over 2 billion individuals worldwide either are or have been infected with HBV.3 Of these, approximately 257 million are chronically infected.2 Approximately 5% to 10% of patients with HBV become chronic carriers.4 Hepatitis B can develop into a chronic disease when acute HBV individuals test positive for the virus for more than 6 months. Chronic infection causes a continuous cycle of hepatocyte damage, leading to an immunologic inflammatory and repair response that can culminate in liver scarring, fibrosis, and cirrhosis.3 Chronic HBV is also a major cause of hepatocellular carcinoma (HCC) and is responsible for over 680,000 deaths, annually.2 Approximately 30% of all cases of cirrhosis and 45% of all cases of HCC are caused by chronic HBV infection.5 Safe and effective vaccination is available that offers a 98% to 100% protection against hepatitis B.1 The presence of the e antigen of the hepatitis B virus (HBeAg) during acute or chronic hepatitis B infection indicates that the virus is replicating; the infected person has high levels of HBV. Spontaneous conversion from e antigen to e antibody is a predictor of long-term clearance of HBV in patients undergoing antiviral therapy, and indicates lower levels of HBV.6 Thus, loss of HBeAg and appearance of antibodies to HBe (aHBe) is a favorable serological marker during acute hepatitis B, indicating the initiation of recovery. Principles of the Procedure The ADVIA Centaur aHBe2 assay is a fully automated 2-step competitive immunoassay using chemiluminescent technology. The Ancillary Well Reagent contains recombinant HBeAg. The Solid Phase contains microparticles coated with aHBe monoclonal antibody. The Lite Reagent contains aHBe monoclonal antibody labeled with acridinium ester. The sample is incubated with the Ancillary Well Reagent. During the incubation period, aHBe in the sample binds with recombinant HBeAg. The Solid Phase and Lite Reagent are added next and bind any recombinant HBeAg not already bound by sample. After a wash step, acid and base are dispensed to initiate the chemiluminescent reaction. An inverse relationship exists between the amount of aHBe activity present in the patient sample and the amount of relative light units (RLUs) detected by the system. A result of reactive or non- reactive is determined according to a cut-off of 1.00 Index established with the calibrators. Refer to Interpretation of Results. 2 / 27 10720831_CP_US_EN Rev. A, DRAFT ADVIA Centaur CP System aHBe2 Reagents Material Description Storage Stability ® ADVIA Centaur aHBe2 ReadyPack primary reagent Unopened at 2–8°C Until expiration date on a pack product Lite Reagent 3.8 mL/reagent pack Onboard 60 days Mouse monoclonal aHBe labelled with acridinium ester in buffer; bovine serum albumin (BSA); mouse IgG; surfactant; preservative Solid Phase 7.5 mL/reagent pack Paramagnetic microparticles coated with mouse monoclonal aHBe in buffer; BSA, surfactant; sodium azide (< 0.1%) Ancillary Well Reagent 3.8 mL/reagent pack Recombinant HBe antigen in buffer; BSA; surfactant; preservative ADVIA Centaur aHBe2 CALa Unopened at 2–8°C Until expiration date on 2.0 mL/vial product Processed human plasma negative and positive for aHBe; sodium azide (< 0.1%) Opened at 2–8°C 60 days At room temperature 8 hours ADVIA Centaur Wash 1a Unopened at 2–25°C Until expiration date on 1500 mL/pack product Phosphate buffered saline; sodium azide (< 0.1%); surfactant Onboard 1 month ADVIA Centaur Wash 1a Unopened at 2–25°C Until expiration date on 2500 mL/pack product Phosphate buffered saline; sodium azide (< 0.1%); surfactant Onboard 1 month a Refer to Storage and Stability. Warnings and Precautions For in vitro diagnostic use. For Professional Use. For Prescription Use Only. CAUTION Federal (USA) law restricts this device to sale by or on the order of a licensed healthcare professional. Safety data sheets (SDS) available on siemens-healthineers.com 10720831_CP_US_EN Rev. A, DRAFT 3 / 27 aHBe2 ADVIA Centaur CP System H317 Warning! P280, May cause an allergic skin reaction. P302+P352, Wear protective gloves/protective clothing/eye protection/face protection. P333+P313, IF ON SKIN: Wash with plenty of soap and water. If skin irritation or rash occurs: Get P362+P364 medical advice/attention. Take off contaminated clothing and wash it before reuse. Contains: 2-methylisothiazol-3(2H)-one (in ADVIA Centaur aHBe2 Lite Reagent, Ancillary Well Reagent). CAUTION POTENTIAL BIOHAZARD Contains human source material. Each donation of human blood or blood component negative for antibodies to HBe antigen was tested by FDA-approved methods for the presence of antibodies to human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2), as well as for hepatitis B surface antigen (HBsAg) and antibody to hepatitis C virus (HCV). The test results were negative (not repeatedly reactive). No test offers complete assurance that these or other infectious agents are absent; this material should be handled using good laboratory practices and universal precautions.7–9 Calibrators and controls contain processed human plasma that is nonreactive for HIV and HCV but is reactive for antibodies to HBe antigen. The units were treated with a BPL-UV inactivation procedure;10 however, all products manufactured using human source material should be handled as potentially infectious. CAUTION This device contains material of animal origin and should be handled as a potential carrier and transmitter of disease. Contains sodium azide as a preservative. Sodium azide can react with copper or lead plumbing to form explosive metal azides. On disposal, flush reagents with a large volume of water to prevent buildup of azides. Disposal into drain systems must be in compliance with prevailing regulatory requirements. Dispose of hazardous or biologically contaminated materials according to the practices of your institution. Discard all materials in a safe and acceptable manner and in compliance with prevailing regulatory requirements. Note For information about reagent preparation, refer to Preparing the Reagents in the Procedure section. Storage and Stability Store all reagents in an upright position, away from light and heat. Do not use products beyond the expiration date printed on the product labeling. For information about product storage and stability, refer to Reagents. Onboard Stability Reagents are stable onboard the system for 60 days. Discard reagents at the end of the onboard stability interval. Do not use products beyond the expiration date printed on the product labeling. The ADVIA Centaur aHBe2 Calibrators are stable on the system for 8 hours. 4 / 27 10720831_CP_US_EN Rev. A, DRAFT ADVIA Centaur CP System aHBe2 Specimen Collection and Handling Serum, EDTA plasma, and lithium heparin plasma are the recommended specimen types for this assay. Do not use heat-inactivated specimens. Collecting the Specimen • Observe universal precautions when collecting specimens. Handle all specimens as if they are capable of transmitting disease.8 • Follow recommended procedures for collection of diagnostic blood specimens by venipuncture.11 • Follow the instructions provided with your specimen collection device for use and processing.12 • Allow blood specimens to clot completely before centrifugation.9 • Keep tubes capped at all times.9 • Test specimens as soon as possible after collecting. Storing the Specimen • Separated specimens are stable for up to 72 hours at room temperature, and for up to 8 days at 2–8°C.