DESCRIPTION OF SYMBOLS USED KIT COMPONENTS SUBSTRATE 1 bottle 1. Avoid microbial contamination of reagents when opening 4. Avoid microbial contamination of the reagents, when Solution of 5-bromo-4-chloro- (100 ml) and removing aliquots from the original vials or bottles. opening and removing aliquots from the original vials or The following are graphical symbols used in or found on MP 3-indolyl phosphate (BCIP) bottles. As this will prematurely reduce the shelf life of the Component Description Quantity Diagnostics products and packaging. These symbols are and nitroblue tetrazolium 2. Do not pipette by mouth. kits and give erroneous results. Use aseptic techniques Provided the most common ones appearing on medical devices and (NBT). including pipettes or disposable pipette tips when drawing their packaging. Some of the common symbols are explained 3. Wear laboratory coats and disposable gloves while aliquots from vials. NITROCELLULOSE STRIPS Available in more detail in the British and European Standard BS EN BLOTTING POWDER 10 packets performing the assay. Discard gloves in biohazard waste- Incorporated with HTLV-I in 18 or 36 980: 2008. Non-fat dry milk (1g each) bags. Wash hands thoroughly afterwards. 5. The kit controls should be assayed concurrently with viral lysate and recombinant strips samples for each test run. Use by Consult Instructions envelope antigens, and a Incubation Tray, 2 or 4 trays 4. It is highly recommended that this assay be performed in Synonyms for this: for Use serum addition control (anti- a biohazard cabinet. Expire Date human IgG) band. Keep dry 9 wells each 6. Use a new pipette tip for each specimen aliquot to prevent cross contamination. HTLV BLOT 2.4 Catalogue and away from light. 5. Keep materials away from food and drink. Batch Code Number Instructions For Use 1 copy WESTERN BLOT ASSAY 7. For best results dispense all reagents while cold and return Synonyms for this are: NON-REACTIVE 1 vial 6. In case of accident or contact with eyes, rinse immediately Instructions For Use Lot Number Forceps to 2°C to 8°C storage as soon as possible. CONTROL (80 µl) 1 pair with plenty of water and seek medical advice. Batch Number Inactivated normal human FOR RESEARCH USE ONLY Caution serum non-reactive for anti- 7. Consult a physician immediately in the event that 8. It is recommended that glassware to be used with the NOT FOR USE IN DIAGNOSTIC PROCEDURES Temperature Limitation Note : Volume of reagents provided are suffi cient for 4 runs. reagents should be washed with 2M hydrochloric acid and HCV, anti-HIV-1/2, anti-HTLV- contaminated materials are ingested or come in contact I/II and HBsAg. Contains with open lacerations or other breaks in the skin. rinsed thoroughly with distilled or deionised water prior to Do not reuse REVISION DATE: 09/10 Note Changes Highlighted sodium azide and thimerosal use. MAK0012-ENG-3 Manufacturer as preservatives. WARNINGS AND PRECAUTIONS 8. Wipe spills of potentially infectious materials immediately 9. Use only reagent grade quality, deionised or distilled water Harmful (Xn) / with absorbent paper and swab the contaminated area with (18 tests kit) : 11082-018 1. For Research Use Only. It is not intended for use in the to dilute reagents. Contains suffi cient Irritant (Xi) STRONG REACTIVE 1 vial 1% sodium hypochlorite solution before work is resumed. (36 tests kit) : 11082-036 diagnosis or prognosis of disease. for <n> tests CONTROL I (80 µl) Sodium hypochlorite should not be used on acid containing Inactivated human serum 2. For Professional use only. spills unless the area is wiped dry with absorbent paper 10. All reagents must be mixed well before use. with high titered antibodies fi rst. Material used (including disposable gloves) should to HTLV-I and non-reactive HEALTH AND SAFETY INFORMATION be disposed as potentially biohazardous material. Do not 11. Working Conjugate solution, Diluted Wash Buffer and CHEMICAL & BIOLOGICAL PRINCIPLES OF THE for anti-HCV, anti-HIV-1/2 autoclave material containing sodium hypochlorite. Blotting Buffer should be prepared fresh prior to use. NAME AND INTENDED USE CAUTION: This kit contains materials of human origin. PROCEDURE and HBsAg. Contains sodium The MP Diagnostics HTLV BLOT 2.4 is a qualitative enzyme azide and thimerosal as No test method can offer complete assurance that 9. Autoclave all used and contaminated materials at 121°C 12. The Working Conjugate solution should be prepared using human blood products will not transmit infection. at 15 p.s.i. for 30 minutes before disposal. Alternatively, immunoassay for the in vitro detection of antibodies to HTLV-I The nitrocellulose strips are incorporated with HTLV-I viral preservatives. a polypropylene container or beaker. and HTLV-II in human serum or plasma samples. This test kit proteins derived from native inactivated disrupted viral particles decontaminate materials in 5% sodium hypochlorite is supplied for Research Use Only. It is not intended for use in STRONG REACTIVE HANDLE ASSAY SPECIMENS, STRONG REACTIVE solution for 30-60 minutes before disposal in biohazard 13. Do not expose reagents or perform test in an area and genetically engineered proteins. Individual nitrocellulose 1 vial AND NON-REACTIVE CONTROLS AS POTENTIALLY the diagnosis or prognosis of disease. CONTROL II waste-bags. containing a high level of chemical disinfectant fumes (e.g. strips are incubated with diluted serum or plasma specimens (80 µl) INFECTIOUS AGENTS. It is recommended that the components and controls. Specifi c antibodies to HTLV-I/II, if present in the Inactivated human serum hypochlorite fumes) during storage or during incubation and test specimens be handled using universal precautions. 10. Decontaminate all used chemicals and reagents by adding specimen will bind to the HTLV-I/II proteins on the strips. The with high titered antibodies They should be disposed of in accordance with established steps. Contact inhibits colour reaction. Also do not expose INTRODUCTION strips are washed to remove unbound materials while antibodies suffi cient volume of sodium hypochlorite to make a fi nal to HTLV-II and non-reactive safety procedures. reagents to strong light. that bind specifi cally to the HTLV proteins can be visualized for anti-HCV, anti-HIV-1/2 concentration of at least 1%. Leave for 30 minutes to The HTLV Blot 2.4 is intended as a supplemental antibody using a series of reactions with goat anti-human IgG conjugated and HBsAg. Contains sodium ensure effective decontamination. 14. The assay should preferably be performed at room assay for Research Use Only. The possible serological profi les The Strong Reactive Control I, Strong Reactive Control II with alkaline phosphatase and the substrate, BCIP/NBT. azide and thimerosal as temperature (25°C ± 3°C). defi ned by the HTLV Blot 2.4 include the following: HTLV and Non-Reactive Control contain Thimerosal and Sodium preservatives. 11. We do not recommend re-use of incubation trays. Seropositive, HTLV-I Seropositive, HTLV-II Seropositive, Azide, the Stock Buffer Concentrate and Wash Buffer 15. Make sure that the test strips are laid with the numbers Seronegative or Indeterminate. Concentrate contain Thimerosal and the Conjugate contains ANALYTICAL PRECAUTIONS LYOPHILIZED STOCK 1 or 2 bottles Sodium azide. Sodium azide can react with copper and lead on the strips facing upwards. BUFFER (each to be used in some plumbing systems to form explosive salts. 1. Optimal assay performance requires STRICT To be reconstituted in reagent reconstituted The quantities used in this kit are small, nevertheless when 16. For Western Blot Assay, it is important to use a rocking ADHERENCE to the assay procedure described in this grade water. Tris buffer with to 100 ml) disposing of azide-containing materials they should be fl ushed platform shaker and not a rotary shaker. Otherwise, Instruction Manual. Deviations from the procedure may heat inactivated animal and away with relatively large quantities of water to prevent metal performance of the kit will be compromised. The lead to aberrant results. non-animal proteins. Contains azide buildup in plumbing system. recommended speed and tilt angle of the shaker are 12 to thimerosal as preservative. 16 cycles per minute, and 5 to 10 degrees, respectively. 2. DO NOT MODIFY OR SUBSTITUTE REAGENTS FROM ONE KIT LOT TO ANOTHER. Controls, conjugate and WASH BUFFER 1 bottle 17. Ensure that automated equipment if used is validated Western Blot strips are matched for optimal performance. CONCENTRATE (20x) (70 ml) before use. Tris with Tween-20. Contains Use only the reagents supplied with the kit. thimerosal as preservative. 18. Ensure that the specimens are added away from the strip. 3. Do not use kit components beyond the expiry date printed Tray can be tilted and specimen added where the buffer is on the kit box. CONJUGATE 1 vial collected at lower end. This prevents dark spot formation Goat anti-human IgG (160 µl) due to specimen addition on the strip. conjugated with alkaline phosphatase. Contains sodium 19. Avoid the use of self-defrosting freezers for the storage of azide as a perservative. reagents and samples. 1 2 3 4. William AE., Fang CT., Slamon DJ. et al. Seroprevalence STORAGE PREPARATION OF REAGENTS Procedure: SUMMARY OF ASSAY PROTOCOLS LIMITATIONS OF THE PROCEDURE and epidemiological correlates of HTLV-I infection in U.S. 1. Using forceps, carefully remove required 1. Store HTLV BLOT 2.4 kit and its components at 2°C to 8°C 1. DILUTED WASH BUFFER Reagents Qty Duration blood donors. Science 1988; 240: 643-646. number of STRIPS from the tube and when not in use. (a) DILUTED WASH BUFFER should be prepared fresh Optimal assay performance requires the strict adherence to the place numbered side up into each well. Nitrocellulose strip 1 - assay procedure described. Deviation from the recommended prior to use.