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In addition to using the MNvit assay to identify chemicals (substances and mixtures) that induce micronuclei, the use of a cytokinesis block, immunochemical labelling of kinetochores, or hybridisation with centromeric/telomeric probes (fluorescence in situ hybridisation (FISH)), also can provide information on the mechanisms of chromosome damage and micronucleus formation (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16).
Okrem použitia skúšky MNvit na identifikáciu chemikálií (látok a zmesí), ktoré indukujú mikrojadrá, môže použitie inhibítorov cytokinézy, imunochemického označovania kinetochorov alebo hybridizácia s centromérovými/telomérovými sondami [fluorescenčná hybridizácia in situ (FISH)] tiež poskytnúť informácie o mechanizmoch poškodenia chromozómov a tvorbe mikrojadier (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16).
The labelling and hybridisation procedures can be used when there is an increase in micronucleus formation and the investigator wishes to determine if the increase was the result of clastogenic and/or aneugenic events.
Postupy označovania a hybridizácie možno použiť, ak dôjde k zvýšenej miere tvorby mikrojadier a výskumník chce určiť, či je tento nárast výsledkom klastogénnych a/alebo aneugénnych procesov.
The use of a DNA specific stain (e.g. acridine orange (61) or Hoechst 33258 plus pyronin-Y (62)) can eliminate some of the artefacts associated with using a non-DNA specific stain.
Použitie špecifického farbiva DNA [napríklad akridínová oranžová (61) alebo Hoechst 33258 plus pyronín-Y (62)] môže eliminovať niektoré umelo vzniknuté štruktúry buniek spojené s používaním farbiva nešpecifického pre DNA.
Anti-kinetochore antibodies, FISH with pancentromeric DNA probes, or primed in situ labelling with pancentromere-specific primers, together with appropriate DNA counterstaining, can be used to identify the contents (chromosome/chromosomal fragment) of micronuclei if mechanistic information of their formation is of interest (15) (16).
Protilátky kinetochoru, FISH (fluorescenčná hybridizácia in situ) s pan-centromerérovými sondami DNA alebo značenie in situ pomocou primerov špecifických pre pancentroméru spolu s vhodným odfarbovacím roztokom DNA možno použiť na identifikáciu obsahov mikrojadier (chromozómy/fragmenty chromozómov), ak sú predmetom záujmu mechanické informácie o ich tvorbe (15) (16).
Other methods for differentiation between clastogens and aneugens may be used if they have been shown to be effective.
Na rozlišovanie medzi klastogénmi a aneugénmi možno použiť aj iné metódy, ak sa ukázalo, že sú účinné.
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In addition, chemical classes or substances containing functional groups shown to act as potential confounders (15) may necessitate the use of guinea pig tests (i.e. B.6; OECD Test Guideline 406 (13)).
Okrem toho chemické triedy alebo látky, ktoré obsahujú potenciálne zmätočné funkčné skupiny (15), si tiež môžu vyžadovať použitie testov s morčatami (t. j. B.6; usmernenie OECD na vykonávanie testov 406) (13).
Limitations that have been identified for the LLNA (1 and Chapter B.42 in this Annex) have been recommended to apply also to the LLNA: BrdU-ELISA (10).
Obmedzenia, ktoré boli identifikované pre LLNA (1 a kapitola B.42 tejto prílohy), sa odporúčajú aj pre LLNA: BrdU-ELISA (10).
Other than such identified limitations, the LLNA: BrdU-ELISA should be applicable for testing any chemicals unless there are properties associated with these chemicals that may interfere with the accuracy of the LLNA: BrdU-ELISA.
Okrem takto identifikovaných obmedzení by sa LLNA: BrdU-ELISA mala uplatňovať pri testovaní akýchkoľvek chemikálií okrem prípadu, keď tieto chemikálie majú vlastnosti, ktoré by mohli narúšať presnosť LLNA: BrdU-ELISA.
In addition, consideration should be given to the possibility of borderline positive results when Stimulation Index (SI) values between 1,6 and 1,9 are obtained (see paragraphs 31-32).
Okrem toho by sa mala zvážiť možnosť hraničných pozitívnych výsledkov v prípade, že sa dosiahnu hodnoty stimulačného indexu (SI) od 1,6 do 1,9 (pozri odseky 31 a 32).
This is based on the validation database of 43 substances using an SI ≥ 1,6 (see paragraph 6) for which the LLNA: BrdU-ELISA correctly identified all 32 LLNA sensitisers, but incorrectly identified two of 11 LLNA non-sensitisers with SI values between 1,6 and 1,9 (i.e. borderline positive) (10).
To je založené na validačnej databáze 43 látok s použitím SI ≥ 1,6 (pozri odsek 6), pre ktoré LLNA: BrdU-ELISA správne identifikovala všetkých 32 látok spôsobujúcich senzibilizáciu podľa LLNA, ale nesprávne identifikovala dve z 11 látok nespôsobujúcich senzibilizáciu podľa LLNA s hodnotami SI od 1,6 do 1,9 (t. j. hraničný pozitívny výsledok) (10).
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