DRG® HBsAg (Confirmatory) ELISAAs o

DRG® HBsAg (Confirmatory) ELISA



As of 24 May 2011 rm (Vers. 1.0)


(EIA-3949 // EIA-5166)



USA:RUO




Please use only the valid version of the package insert provided with the kit.


This kit is intended for Research Use Only.


Not for use in diagnostic procedures.


1. INTENDED USE

A simple procedure based on an immunoreaction of neutralization is used in combination with the HBsAg assay.

2. PRINCIPLE OF THE ASSAY
The device has to be used in combination with the product code EIA-4545 or EIA-4134 for the determination of HBsAg in human sera and plasma.
The sample, whose repeatedly positivity for HBsAg has to be confirmed, is premixed with a reagent containing high titer anti HBsAg antibodies that will neutralize the antigen is really present in the sample.
The neutralized sample is then tested for HBsAg according to the procedure reported for the specific device.
If the positivity in the first screening test is specifically related to the presence of HBsAg in the sample, the same will not react any more in the assay having been neutralized by the antibody.
If at contrary the positivity of the sample is not abolished by the neutralization reaction, this reactivity is not due specifically to the presence of HBsAg in the sample, but to some interfering substance.

3. CONTENT OF THE KIT
The set contains the following reagents:

a. Neutralizing Reagent SOLN NEUT It contains high titer human plasma positive for anti HBsAg antibodies, 0.2 mg/ml gentamicine sulphate and 0.1% Kathon GC as preservatives.

b. Control Reagent CONTROL

It contains human plasma negative for anti HBsAg antibodies, 0.2 mg/ml gentamicine sulphate and 0.1% Kathon GC as preservatives.

c. Assay Diluent DILSPE
0.15 M NaCl phosphate buffered solution pH 7.0 + 0.2 containing 0.1% Kathon GC for the dilution of over ranging samples.
Note: Reagents have been tested and found negative for HBsAg, HCV Ab and HIV Ab with CE-marked kits.










DRG International, Inc., USA Fax: [phone] e-mail: [e-mail] 1




DRG® HBsAg (Confirmatory) ELISA



As of 24 May 2011 rm (Vers. 1.0)


(EIA-3949 // EIA-5166)



USA:RUO







umber of tests ode
ontrol Reagent (CONTROL) Neutralizing Reagent
(SOLN NEUT) Phosphate Buffered Saline (DILSPE)
Package insert

20 EIA 949
1x4ml/vial

1x4ml/vial

1x30ml/vial

N° 1

40 EIA- 166
1x8ml/vial

1x8ml/vial

1x60ml/vial

N° 1



4. MATERIALS REQUIRED BUT NOT PROVIDED
1. Device for HBsAg determination, name HBsAg One, code EIA-3949 or EIA-5166. 2. Isotonic sterile solution.
3. Calibrated Micropipettes and disposable plastic tips. 4. Timer with 60 minute range or higher.
5. Absorbent paper tissues.
6. Calibrated ELISA microplate thermostatic incubator (dry or wet), capable to provide shaking at 1300 rpm+/-150, set at +37°C.
7. Calibrated ELISA microwell reader with 450nm (reading) and possibly with 620-630nm (blanking) filters. 8. Calibrated ELISA microplate washer.
9. Vortex or similar mixing tools.
10. Disposable plastic tube of 2-5 ml.





















DRG International, Inc., USA Fax: [phone] e-mail: [e-mail] 2




DRG® HBsAg (Confirmatory) ELISA



As of 24 May 2011 rm (Vers. 1.0)


(EIA-3949 // EIA-5166)



USA:RUO




5. WARNINGS AND PRECAUTIONS
1. 2. The Set has to be used by skilled and properly trained technical personnel only, under the supervision of a medical doctor responsible of the laboratory.
3. When the device is used for confirmation of a sample repeatedly positive in the screening of blood units and blood components, it has to be used in a laboratory certified and qualified by the national authority in that field (Ministry of Health or similar entity) to carry out this type of analysis.
4. The laboratory environment should be controlled so as to avoid contaminants such as dust or air-born microbial agents, when opening the vials contained in the set.
5. Upon receipt, store the kit at 2..8°C into a temperature controlled refrigerator or cold room.
6. Do not interchange Reagents between different lots of the device. It is even recommended that Reagents between two sets of the same lot are not interchanged.
7. Check that the Reagents of the device are clear and do not contain visible heavy particles or aggregates. If not, advise the laboratory supervisor to initiate the necessary procedures for kit replacement.
8. Avoid cross-contamination between kit reagents by using disposable tips and changing them between the use of each one.
9. Do not use the set after the expiration date stated on the external container and internal (vials) labels.
10. Treat all specimens as potentially infective. All human serum specimens should be handled at Biosafety Level 2, as recommended by the Center for Disease Control, Atlanta, U.S. in compliance with what reported in the Institutes of Health’s publication: “Biosafety in Microbiological and Biomedical Laboratories”, ed. 1984.
11. Waste produced during the use of the set in combination with the devise for HBsAg determination has to be discarded in compliance with national directives and laws concerning laboratory waste of chemical and biological substances. In particular, liquid waste generated from the washing procedure, from residuals of controls and from samples has to be treated as potentially infective material and inactivated before waste. Suggested procedures of inactivation are treatment with a 10% final concentration of household bleach for 16-18 hrs or heat inactivation by autoclave at 121°C for 20 min..
12. Accidental spills from samples and operations have to be adsorbed with paper tissues soaked with household bleach and then with water. Tissues should then be discarded in proper containers designated for laboratory/hospital waste.
13. Other waste materials generated from the use of the kit (example: tips used for samples and controls, used microplates) should be handled as potentially infective and disposed according to national directives and laws concerning laboratory wastes.
14. Refer to the Instructions for Use of the product code EIA-3949, EIA-5166 used in combination for the confirmation assay.


6. SPECIMEN: PREPARATION AND WARNINGS
1. The sample turned out to be repeatedly positive in the first HBsAg determination with HBsAg One has to be used for the test of neutralization. Treat the sample as described in section L.
2. Avoid any addition of preservatives to samples after first screening; especially sodium azide as this chemical would affect the enzymatic activity of the conjugate, generating false negative results.
3. Samples have to be clearly identified with codes or names in order to avoid misinterpretation of results.
4. Haemolysed (red) and lipemic (“milky”) samples have to be discarded by definition anyway as they could generate false results in the test for HBsAg.
5. Samples containing residues of fibrin or heavy particles or microbial filaments and bodies should be discarded as well as they could give rise to false positive results both in HBsAg first assay and even in the confirmation one.

DRG International, Inc., USA Fax: [phone] e-mail: [e-mail] 3




DRG® HBsAg (Confirmatory) ELISA



As of 24 May 2011 rm (Vers. 1.0)


(EIA-3949 // EIA-5166)



USA:RUO




6. The assay is not suitable to confirm the negativity of samples that turned out to be negative in the first HBsAg screening test.
7. Sera and plasma can be stored at +2°..8°C for up to five days after collection. For longer storage periods, samples can be stored frozen at –20°C for several months.
8. Any frozen sample should not be frozen/thawed more than once as this may generate particles that could affect the test result. If some turbidity is present or presence of micro particles is suspected after thawing, filter the sample on a disposable 0.2-0.8u filter to clean it up for testing or use the two-steps alternative method.
9. Refer to the Instructions for Use of the product code EIA-3949, EIA-5166 used in combination for the confirmation assay.






































DRG International, Inc., USA Fax: [phone] e-mail: [e-mail] 4




DRG® HBsAg (Confirmatory) ELISA



As of 24 May 2011 rm (Vers. 1.0)


(EIA-3949 // EIA-5166)



USA:RUO





7. INSTRUMENTS AND TOOLS USED IN COMBINATION WITH THE KIT

1. Micropipettes have to be calibrated to deliver the correct volume required by the assay and must be submitted to regular decontamination (70% ethanol, 10% solution of bleach, hospital grade disinfectants) of those parts that could accidentally come in contact with the sample or the components of the kit. They should also be regularly maintained in order to show a precision of 1% and a trueness of +2%.
2. The ELISA incubator has to be set at +37°C (tolerance of +1°C) and regularly checked to ensure the correct temperature is maintained. Both dry incubators and water baths are suitable for the incubations, provided that the instrument is validated for the incubation of ELISA tests.
3. In case of shaking during incubations, the instrument has to ensure 350 rpm +150. Amplitude of shaking is very important as a wrong one could give origin to splashes and therefore to some false positive result.
4. The ELISA washer is extremely important to the overall performances of the assay. The washer must be carefully validated and correctly optimized using the kit controls/calibrator and reference panels, before using the kit for routine laboratory tests. Usually 4-5 washing cycles (aspiration + dispensation of 350ul/well of washing solution = 1 cycle) are sufficient to ensure that the assay performs as expected. A soaking time of 20-30 seconds between cycles is suggested. In order to set correctly their number, it is recommended to run an assay with the kit controls/calibrator and well-characterized negative and positive reference samples, and check to match the values reported below in the section “Internal Quality Control”. Regular calibration of

Kết quả (Việt) 1:

DRG ® HBsAg (Confirmatory) ELISATheo 24 có thể 2011 rm (Vers. 1.0) (DỰ ÁN-3949 / / DỰ ÁN-5166)HOA KỲ: RUO Xin vui lòng sử dụng chỉ có phiên bản hợp lệ của chèn gói cung cấp với các bộ.Kit này được thiết kế để nghiên cứu sử dụng chỉ.Không để sử dụng trong thủ tục chẩn đoán.1. MỤC ĐÍCH SỬ DỤNGMột thủ tục đơn giản dựa trên một immunoreaction trung hòa được sử dụng kết hợp với các khảo nghiệm HBsAg.2. NGUYÊN TẮC CỦA CÁC KHẢO NGHIỆMThiết bị đã được sử dụng kết hợp với mã sản phẩm dự án-4545 hoặc dự án-4134 xác định HBsAg trong huyết thanh của con người và plasma.Mẫu, mà liên tục tích cực cho HBsAg đã được xác nhận, là premixed với một tinh khiết có cao titer chống kháng thể HBsAg sẽ vô hiệu hóa các kháng nguyên là thực sự hiện diện trong mẫu.Neutralized mẫu sau đó được thử nghiệm cho HBsAg theo thủ tục báo cáo cho thiết bị cụ thể.Nếu dương tính trong xét nghiệm sàng lọc đầu tiên đặc biệt là liên quan đến sự hiện diện của HBsAg trong mẫu, như vậy sẽ không phản ứng bất kỳ chi tiết trong khảo nghiệm có được vô hiệu hóa do kháng thể.Nếu tại ngược lại tích cực của mẫu vật không bị xoá bỏ bằng phản ứng trung hòa, phản ứng này không phải là do cụ thể đến sự hiện diện của HBsAg trong mẫu, mà một số chất interfering.3. NỘI DUNG CỦA KITBộ chứa các chất phản ứng sau:a. vô hiệu hóa tinh khiết SOLN NEUT có titer cao huyết tương của con người tích cực cho chống kháng thể HBsAg, cách 0.2 mg/ml gentamicine sulphate và 0.1% Kathon GC như là chất bảo quản.b. Control Reagent CONTROLIt contains human plasma negative for anti HBsAg antibodies, 0.2 mg/ml gentamicine sulphate and 0.1% Kathon GC as preservatives.c. Assay Diluent DILSPE0.15 M NaCl phosphate buffered solution pH 7.0 + 0.2 containing 0.1% Kathon GC for the dilution of over ranging samples.Note: Reagents have been tested and found negative for HBsAg, HCV Ab and HIV Ab with CE-marked kits.DRG International, Inc., USA Fax: [phone] e-mail: [e-mail] 1 DRG® HBsAg (Confirmatory) ELISAAs of 24 May 2011 rm (Vers. 1.0) (EIA-3949 // EIA-5166)USA:RUO umber of tests odeontrol Reagent (CONTROL) Neutralizing Reagent(SOLN NEUT) Phosphate Buffered Saline (DILSPE)Package insert 20 EIA 9491x4ml/vial1x4ml/vial1x30ml/vialN° 1 40 EIA- 1661x8ml/vial1x8ml/vial1x60ml/vialN° 1 4. MATERIALS REQUIRED BUT NOT PROVIDED1. Device for HBsAg determination, name HBsAg One, code EIA-3949 or EIA-5166. 2. Isotonic sterile solution.3. Calibrated Micropipettes and disposable plastic tips. 4. Timer with 60 minute range or higher.5. Absorbent paper tissues.6. Calibrated ELISA microplate thermostatic incubator (dry or wet), capable to provide shaking at 1300 rpm+/-150, set at +37°C.7. Calibrated ELISA microwell reader with 450nm (reading) and possibly with 620-630nm (blanking) filters. 8. Calibrated ELISA microplate washer.9. Vortex or similar mixing tools.10. Disposable plastic tube of 2-5 ml.DRG International, Inc., USA Fax: [phone] e-mail: [e-mail] 2 DRG® HBsAg (Confirmatory) ELISAAs of 24 May 2011 rm (Vers. 1.0) (EIA-3949 // EIA-5166)USA:RUO 5. WARNINGS AND PRECAUTIONS1. 2. The Set has to be used by skilled and properly trained technical personnel only, under the supervision of a medical doctor responsible of the laboratory.3. When the device is used for confirmation of a sample repeatedly positive in the screening of blood units and blood components, it has to be used in a laboratory certified and qualified by the national authority in that field (Ministry of Health or similar entity) to carry out this type of analysis.4. The laboratory environment should be controlled so as to avoid contaminants such as dust or air-born microbial agents, when opening the vials contained in the set.5. Upon receipt, store the kit at 2..8°C into a temperature controlled refrigerator or cold room.6. Do not interchange Reagents between different lots of the device. It is even recommended that Reagents between two sets of the same lot are not interchanged.7. Check that the Reagents of the device are clear and do not contain visible heavy particles or aggregates. If not, advise the laboratory supervisor to initiate the necessary procedures for kit replacement.8. Avoid cross-contamination between kit reagents by using disposable tips and changing them between the use of each one.9. Do not use the set after the expiration date stated on the external container and internal (vials) labels.10. Treat all specimens as potentially infective. All human serum specimens should be handled at Biosafety Level 2, as recommended by the Center for Disease Control, Atlanta, U.S. in compliance with what reported in the Institutes of Health’s publication: “Biosafety in Microbiological and Biomedical Laboratories”, ed. 1984.11. Waste produced during the use of the set in combination with the devise for HBsAg determination has to be discarded in compliance with national directives and laws concerning laboratory waste of chemical and biological substances. In particular, liquid waste generated from the washing procedure, from residuals of controls and from samples has to be treated as potentially infective material and inactivated before waste. Suggested procedures of inactivation are treatment with a 10% final concentration of household bleach for 16-18 hrs or heat inactivation by autoclave at 121°C for 20 min..12. Accidental spills from samples and operations have to be adsorbed with paper tissues soaked with household bleach and then with water. Tissues should then be discarded in proper containers designated for laboratory/hospital waste.13. Other waste materials generated from the use of the kit (example: tips used for samples and controls, used microplates) should be handled as potentially infective and disposed according to national directives and laws concerning laboratory wastes.14. Refer to the Instructions for Use of the product code EIA-3949, EIA-5166 used in combination for the confirmation assay.


6. SPECIMEN: PREPARATION AND WARNINGS
1. The sample turned out to be repeatedly positive in the first HBsAg determination with HBsAg One has to be used for the test of neutralization. Treat the sample as described in section L.
2. Avoid any addition of preservatives to samples after first screening; especially sodium azide as this chemical would affect the enzymatic activity of the conjugate, generating false negative results.
3. Samples have to be clearly identified with codes or names in order to avoid misinterpretation of results.
4. Haemolysed (red) and lipemic (“milky”) samples have to be discarded by definition anyway as they could generate false results in the test for HBsAg.
5. Samples containing residues of fibrin or heavy particles or microbial filaments and bodies should be discarded as well as they could give rise to false positive results both in HBsAg first assay and even in the confirmation one.

DRG International, Inc., USA Fax: [phone] e-mail: [e-mail] 3




DRG® HBsAg (Confirmatory) ELISA



As of 24 May 2011 rm (Vers. 1.0)


(EIA-3949 // EIA-5166)



USA:RUO




6. The assay is not suitable to confirm the negativity of samples that turned out to be negative in the first HBsAg screening test.
7. Sera and plasma can be stored at +2°..8°C for up to five days after collection. For longer storage periods, samples can be stored frozen at –20°C for several months.
8. Any frozen sample should not be frozen/thawed more than once as this may generate particles that could affect the test result. If some turbidity is present or presence of micro particles is suspected after thawing, filter the sample on a disposable 0.2-0.8u filter to clean it up for testing or use the two-steps alternative method.
9. Refer to the Instructions for Use of the product code EIA-3949, EIA-5166 used in combination for the confirmation assay.






































DRG International, Inc., USA Fax: [phone] e-mail: [e-mail] 4




DRG® HBsAg (Confirmatory) ELISA



As of 24 May 2011 rm (Vers. 1.0)


(EIA-3949 // EIA-5166)



USA:RUO





7. INSTRUMENTS AND TOOLS USED IN COMBINATION WITH THE KIT

1. Micropipettes have to be calibrated to deliver the correct volume required by the assay and must be submitted to regular decontamination (70% ethanol, 10% solution of bleach, hospital grade disinfectants) of those parts that could accidentally come in contact with the sample or the components of the kit. They should also be regularly maintained in order to show a precision of 1% and a trueness of +2%.
2. The ELISA incubator has to be set at +37°C (tolerance of +1°C) and regularly checked to ensure the correct temperature is maintained. Both dry incubators and water baths are suitable for the incubations, provided that the instrument is validated for the incubation of ELISA tests.
3. In case of shaking during incubations, the instrument has to ensure 350 rpm +150. Amplitude of shaking is very important as a wrong one could give origin to splashes and therefore to some false positive result.
4. The ELISA washer is extremely important to the overall performances of the assay. The washer must be carefully validated and correctly optimized using the kit controls/calibrator and reference panels, before using the kit for routine laboratory tests. Usually 4-5 washing cycles (aspiration + dispensation of 350ul/well of washing solution = 1 cycle) are sufficient to ensure that the assay performs as expected. A soaking time of 20-30 seconds between cycles is suggested. In order to set correctly their number, it is recommended to run an assay with the kit controls/calibrator and well-characterized negative and positive reference samples, and check to match the values reported below in the section “Internal Quality Control”. Regular calibration of