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Reagents for Use with the PREVI® Color Gram 14028 E - En - 2015/03

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Reagents for Use with the PREVI® Color Gram 14028 E - En - 2015/03 29519 to 29524 and 29582 to 29587 14028 E - en - 2015/03 ® Reagents for use with the PREVI Color Gram The PREVI® Color Gram dyes are used to stain micro-organisms according to Gram’s method. ® They are intended for use exclusively with the PREVI Color Gram instrument. SUMMARY AND EXPLANATION COMPOSITION The PREVI® Color Gram dyes are used with the PREVI® Theoretical formula for 1 liter: Color Gram instrument to stain bacterial and fungal micro- RA1 ACETONE SAFRANIN solution * organisms for direct examination, according to Gram’s Safranin 0 (CAS no. 477-73-6 – CI (Color index) no. 50240) ............ 2.5 g method. Buffer ................................................................................................... 0.8 g ® ® The PREVI Color Gram instrument sprays the PREVI Acetone / Alcohol .................................................................................... 1 l Color Gram reagents onto microscope slides which have RA2 SAFRANIN solution ** been smeared with specimens to be examined. The Safranin 0 (CAS no. 477-73-6 – CI no. 50240) .................................. 2.5 g staining procedure is rapid and precise. Buffer ................................................................................................... 0.8 g Alcohol .................................................................................................... 1 l Each reagent has a specific position (A, B, C, D, E) on the ® PREVI Color Gram platform that is also indicated on the RA3 ACETONE FUCHSIN solution * Basic Fuchsin (CAS no. 569-61-9 – CI no. 42500) ............................... 1 g reagent label. Buffer ................................................................................................... 0.8 g Acetone / Alcohol .................................................................................... 1 l PRINCIPLE Specimen fixation can be performed either manually (heat RA4 FUCHSIN solution ** ® Basic Fuchsin (CAS no. 569-61-9 – CI no. 42500) ............................... 1 g or alcohol) or using the PREVI Color Gram instrument Buffer ................................................................................................... 0.8 g (ethanol or methanol) (E). Alcohol .................................................................................................... 1 l The crystal violet (C) rapidly penetrates the microbial cells RB IODINE solution and stains them blue-black. The excess dye is washed Lugol .................................................................................................... 13 g away with distilled water (D) in order to avoid the Polyvinylpyrrolidone ............................................................................. 26 g formation of precipitate on the outside of the colored cells. Demineralized water ............................................................................... 1 l The polyvinylpyrrolidone-stabilized iodine (mordant) (B) RC CRYSTAL VIOLET solution penetrates the microbial cell and complexes with the Crystal violet (CAS no. 548-62-9 – CI no. 42555) .............................. 1.5 g crystal violet. Alcohol ............................................................................................... 0.02 l Preservatives ....................................................................................... 0.1g The excess iodine is washed away with distilled water (D). Demineralized water ............................................................................... 1 l The solution in position A (RA1, RA2, RA3 or RA4) contains the decolorizer and counterstain. * Signal word : DANGER During decolorization (A), the iodine-crystal violet complex is eliminated for Gram-negative organisms only, due to the chemical composition of the cell wall. The safranin or fuchsin (A) then acts as a counterstain. Only Gram-negative organisms stain red-pink. Gram- positive organisms that have not been decolorized remain Hazard statements blue-black. After staining has been performed, the PREVI® Color EUH066 : Repeated exposure may cause skin dryness or Gram dries the slides. cracking. H225 : Highly flammable liquid and vapour. CONTENT OF THE KIT H319 : Causes serious eye irritation. H336 : May cause drowsiness or dizziness. Individual packs: Precautionary statements 29519 or 29582 RA1 = 1 x 500ml P210 : Keep away from heat, hot surfaces, sparks, open 29520 or 29583 RA2 = 1 x 500ml flames and other ignition sources. No smoking. P233 : Keep container tightly closed. 29521 RA3 = 1 x 500ml P280 : Wear protective gloves/protective clothing/eye protection/face protection. 29522 or 29585 RA4 = 1 x 500ml P305 + P351 + P338 : IF IN EYES: Rinse cautiously with 29523 or 29586 RB = 1 x 500ml water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. 29524 or 29587 RC = 1 x 500ml P337 + P313 : If eye irritation persists: Get medical advice/attention. bioMérieux SA English - 1 Reagents for use with the PREVI® Color Gram 14028 E - en - 2015/03 ** Signal word : DANGER WARNINGS AND PRECAUTIONS • For in vitro diagnostic use only. • For professional use only. • Refer to the hazard statements “H” and precautionary statements “P” indicated above. Hazard statements • All specimens, microbial cultures and inoculated H225 : Highly flammable liquid and vapour. products should be considered infectious and handled H301 : Toxic if swallowed. appropriately. Aseptic technique and usual precautions H311 : Toxic in contact with skin. for handling the bacterial group studied should be H319 : Causes serious eye irritation. observed throughout this procedure. Refer to "CLSI H331 : Toxic if inhaled. M29-A, Protection of Laboratory Workers From H336 : May cause drowsiness or dizziness. Occupationally Acquired Infections; Approved Guideline H370 : Causes damage to organs. – Current Revision". For additional information on handling precautions, refer to "Biosafety in Precautionary statements Microbiological and Biomedical Laboratories – CDC/NIH P210 : Keep away from heat, hot surfaces, sparks, open – Latest edition", or the current regulations in the flames and other ignition sources. No smoking. country of use. P280 : Wear protective gloves/protective clothing/eye • The reagents required but not provided (ethanol or protection/face protection. methanol) may present a chemical risk and should be P301 + P312 : IF SWALLOWED: Call a POISON handled with care. Refer to the safety data provided by CENTER/doctor if you feel unwell. the original manufacturer. P302 + P352 : IF ON SKIN: Wash with plenty of water. • As a precaution in case of a leak or overflow, P304 + P340 : IF INHALED: Remove person to fresh air bioMérieux recommends placing the bottle, that can and keep comfortable for breathing. contain up to 5 liters of methanol or ethanol, in a metal P305 + P351 + P338 : IF IN EYES: Rinse cautiously with drip tray. Check local regulations concerning the water for several minutes. Remove contact lenses, if maximum authorized volume of alcohol that can be present and easy to do. Continue rinsing. stored in a laboratory. P337 + P313 : If eye irritation persists: Get medical • Do not use reagents after the expiry date. advice/attention. • Before use, make sure the tamper-proof seal on the For further information, consult the safety data sheet. bottle screw-caps is intact. • Caution: It is important to respect the position of the ® MATERIAL REQUIRED BUT NOT PROVIDED reagents on the PREVI Color Gram platform. For this purpose, the letters that figure on the reagent bottle • ® PREVI Color Gram Instrument labels (A, B, C, D or E) correspond to the positions of • ® D&E: empty 5-liter bottles for ethanol or methanol and the reagents on the PREVI Color Gram platform. distilled water • Interpretation of test results should be made taking into • D&E: empty 500ml bottles for ethanol or methanol and consideration the patient’s history, and, if necessary, the distilled water results of any other tests performed. • Distilled water • The performance data presented were obtained using • ® Ethanol or methanol (see specifications in the PREVI the procedure indicated in this package insert. Any Color Gram User’s Manual) change in the procedure may affect the results. • Clean and grease-free glass slides • Thermal block STORAGE CONDITIONS • Immersion oil • Microscope with oil-immersion objective • Store the dyes at 15-25°C until the expiry date. • If the dyes need to be removed from the instrument, put the original caps back on the bottles and store them between 15 and 25°C in the dark. bioMérieux SA English - 2 Reagents for use with the PREVI® Color Gram 14028 E - en - 2015/03 SPECIMENS With the specimens, the PREVI® Color Gram Human specimens (sputum, vaginal swabs, blood decolorization programs 2 and 3 gave equivalent or better cultures, bronchoalveolar liquid, cerebrospinal fluid, feces results than the manual staining method in respectively urine) fixed on microscope slides can be used, and also 99.14% and 100% of cases, and the 2 decolorization programs were equivalent in 99.15% of cases. Moreover, strains isolated from a culture. ® Good laboratory practices for collection and transport staining obtained for slides stained using the PREVI should be respected and adapted to the type of specimen. Color Gram was more distinctive; the regularity and the Any other type of specimen must be validated by the user. Gram-positive / Gram-negative contrast of the staining were slightly better than with the manual staining method. Internal validation showed equivalent performance for all INSTRUCTIONS
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