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VITOTOX

The VitotoxTM test is a high throughput bacterial genotoxicity assay which can detect DNA damage caused by genotoxic compounds as light emission changes depending on SOS DNA repair induction.

Use of the Vititox test:

The Vitotox test was developed to test chemical substances on genotoxity and citytoxity. Vitotox is a licensed and trade marked product.

This test is performed in the start of pipeline for developing new pharmaceutical substances. The test, carried out in 1 day, is already used by some of the most important pharmaceutical product developers in the world. Up till now, these companies used the more complicated Ames test, witch takes 3 days to have results. Usually, the Ames test is done, at the end of the pharmaceutical developing pipeline.
A sublicense for continuous use can be subject of co-operating business with Gentaur Institute.

The use of this test is not limited to pharmaceutical research. For every interest in DNA-damaging substances, this test could be useful. Further research on it will proceed during the next decade.
It can be in the environmental, medical or biological fields of research.

Up till now the test proves to perform by excellence on purified substances. Non-purified mixes can give false positive results due to internal factor interfering of the substance mix.

 If this is what you need, the test will give you fast and reliable results.

 
For more information, you can find www.gentaur.com or you can take contact on [email protected].  

 Protocol for Vitotox (Version Français)

Overview of the reagents and equipment for doing a vitotox test.

1. A Vitotox 10 Kit containing: (*)

• 5 vials (0.1mL yellow label) with Salmonella typhimurium TA104 recN2-4 (Genox strain).
  
  These carry a plasmid containing the bacterial luciferase operon (luxCDABE) of luminous bacteria Vibrio fischeri under transcriptional control of an recN promoter (recN2-4). The latter strain constitutively expresses the lux operon. Genotoxic compounds activate the recN promoter in the Genox strain, which results in transcriptional induction of the lux operon followed by the enhancement of light emission.
   

• 5 vials (0.1mL blue label) with Salmonella typhimurium TA104 pr1 (Cytox strain).
  These are to identify the non-specific enhancement of light emission. Concomitant use of the Genox and the Cytox strains allows us to identify false positive results caused by non-specific light emission emission induced by other mechanisms, and not by the genotoxic effect.
   

•    10 vials (10mL white label) Rich growth medium with enhancer.
   

•     5 vials ( 8 mL white label) Poor growth medium.
   

•     5 dark 96 well plates
   

•     3 clear 96 well plates
   

•     20 reagent basins.
   

•     1 catalog of the Vitotox 10 kit, describing the test. [1]
   

•     1 overview document about what you need to proceed the tests with all references.
   

•     1 copy of the quality control report for the Genox and Cytox strains.

(*) This Vitotox 10 Kit is purchased from GENTAUR Molecular Products. A Vitotox-384 kit for use with 384-well microplates can be assembled for you on request.

2. S9 extract and Co-factor I (additionally needed and not included in the test Vitotox 10 kit).

*   Rat liver S9, is prepared as a 9000x g supernatant fraction of the liver homogenates prepared from male SD rats treated with phenobarbital and 5,6-benzoflavone.
    (Rat liver S9 can be purchased from Kikkoman Co. (Chiba, Japan)

*   Co-factor I to prepare S9 mix solutions at the volume ratio of 1:9 (Co-factor can be purchased from Oriental Yeast Co., Ltd (Tokyo, Japan).

*   Your compounds that you want to test.
     Remark: Any preliminary preparing, separation or purification, that needs to be done, to get satisfactory results.
     (Gentaur can give you advice and support on these matters. Please send your requests to [email protected]).

 3. Reference chemical substances for testing (facultative).

Several test compounds were tested during the development research program at the research institute in Belgium.

GENTAUR Molecular Products can supply some or more of these substances on request.
The use of these is facultative, not really needed for primary testing of your interested compounds. It can be usefull for an additional or second validation of the Vitotox test.

Table 1 in the catalog {1} gives the results of the Vitotox test in comparison with other genotoxicity tests, such as the Ames test, using 59 carcinogens and non-carcinogens     mainly evaluated in the National Toxicology Program (NTP), USA. (ref.[1] in the catalogue {1}).
During the research program, 61 chemicals were selected as test substances, witch have already been examined for their mutagenicity with the Ames test.

(see table 1 in the catalogue {1}).

The chemical compounds were purchased from:

        Wako Pure Chemical Ind. Ltd. (Tokio, Japan),

        Tokyo Kasei Kogyo Co. Ltd. (Tokio, Japan),

        Nacalai Tesque Inc. (Kyoto, Japan),

        and Sigma-Aldrich (St. Louis, U.S.A.).

Our 137 pharmaceutical candidates were synthesized in the developing research institute.
The developing research institute for Vitotox co-operated with the

Toxicology Laboratory, Pharmaceuticals Research Unit, Research & Development Division, Mitsubishi Pharma Corporation, 1-1-1, Kazusakamatari, Kisarazu, Chiba 292-0818, Japan

 4. Equipment that is needed in your laboratory to perform the Vitotox 10 kit test.

Gentaur Molecular Products can supply the needed equipment, mentioned here under.
Please send your inquiry to [email protected]. This equipment is in our catalogs on www.gentaur.com. Don't hesitate to contact at [email protected] for an offer.

A specific complete catalogue with technical description for all the Vito ox-related equipment is being made now. We expect it to be ready from March 2008.

Equipment list:

            Luminometer and Software for automatic calculation of Genox/Cytox ratio.

            Orbital Shaker with temperature control 35 - 37°C

            Photometer 590 nm

            12 channel pipette 5 - 50 µL

            12 channel pipette 50 - 300 µL

            Single channel pipettes 10 - 100 µL, 20 - 200 µL, 1 - 5 mL

 5. Training, inter scientist relations and congress participation.

• GENTAUR Molecular Products can intermediate for a 1 day training at the research institute in Belgium.  

Such a training can be useful to see how a Vitotox test is carried out, and how the results of it can be interpreted in a conclusion about the toxicity, according to the scientific standards.
On a simple request, we will send you an offer with prices and conditions.

•  GENTAUR Molecular Products can intermediate for inter scientists contacts with scientists who are involved in the development of the Vitotox science.

Because bio-science is moving fast, you might be interested to make contact with scientists who work on the field of Genetic toxicity and Mutagen activity of substances.
For contacts we will help you to achieve direct and suitable communication channels.

•  GENTAUR Molecular Products can inform you about congresses and literarature concerning genotoxicity by sending you our Quarterly Journal "GENTAUR NEWS".

Please inform us, if you want to be informed. We will put you on our mailing list.

More info 
Discontinuation Letter

Copyright © 2002 GENTAUR Molecular Products
Last modified: 05/29/09