Bioluminescent Salmonella reverse mutation assay: a screen for detecting mutagenicity with high throughput attributes.

Here, we describe the development and evaluation of a novel bioluminescent high-throughput Salmonella reverse mutation assay applicable to the screening of large numbers of small molecules. The bioluminescent Salmonella assay utilizes genetically engineered standard Salmonella tester strains TA98 and TA100 expressing the lux(CDABE) operon from Xenorhabdus luminescence. In principle, the assay employs bioluminescence as a sensor of changes in bacterial metabolism associated with starvation or energy depletion effectively identifying colonies of histidine-independent revertant cells in a high-throughput fashion. The assay provides highly concordant data with the outcome in the standard Salmonella plate incorporation reverse mutation assay. Since the results of the standard Salmonella plate assay are required by various regulatory agencies for approval of new drugs, the bioluminescent Salmonella assay can be effectively used for prioritization of compounds in pharmaceutical drug discovery as well as the evaluation of environmental and industrial chemicals. Because of its high throughput attributes, the assay permits effective, fast and economical screening of a large series of structural analogs enabling the investigation of structure-activity relationships.

[1]  K. Nealson,et al.  Cloning, organization, and expression of the bioluminescence genes of Xenorhabdus luminescens , 1990, Journal of bacteriology.

[2]  T. Brooks.  The use of a streamlined bacterial mutagenicity assay, the MINISCREEN. , 1995, Mutagenesis.

[3]  B. Ames,et al.  Carcinogens are mutagens: a simple test system combining liver homogenates for activation and bacteria for detection. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[4]  L. Verschaeve,et al.  The VITOTOX test, an SOS bioluminescence Salmonella typhimurium test to measure genotoxicity kinetics. , 1997, Mutation research.

[5]  J W Green,et al.  A review of the genotoxicity of marketed pharmaceuticals. , 2001, Mutation research.

[6]  Bonnie L Bassler,et al.  Small Talk Cell-to-Cell Communication in Bacteria , 2002, Cell.

[7]  B. Ames,et al.  Detection and classification of mutagens: a set of base-specific Salmonella tester strains. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[8]  J. DiMasi,et al.  Success rates for new drugs entering clinical testing in the United States , 1995, Clinical pharmacology and therapeutics.

[9]  T Sofuni,et al.  ICH-harmonised guidances on genotoxicity testing of pharmaceuticals: evolution, reasoning and impact. , 1999, Mutation research.

[10]  B. Ames,et al.  Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. , 1975, Mutation research.

[11]  B. Ames,et al.  Revised methods for the Salmonella mutagenicity test. , 1983, Mutation research.

[12]  E. Zeiger,et al.  Salmonella mutagenicity tests: III. Results from the testing of 255 chemicals. , 1987, Environmental mutagenesis.

[13]  E. Zeiger,et al.  Salmonella mutagenicity tests: II. Results from the testing of 270 chemicals. , 1986, Environmental mutagenesis.

[14]  P. Williams,et al.  lux genes and the applications of bacterial bioluminescence. , 1992, Journal of general microbiology.

[15]  E. Zeiger,et al.  Salmonella mutagenicity test results for 250 chemicals. , 1983, Environmental mutagenesis.

[16]  B. Margolin,et al.  Prediction of rodent carcinogenicity utilizing a battery of in vitro and in vivo genotoxicity tests , 1999, Environmental and molecular mutagenesis.

[17]  E. Zeiger,et al.  Salmonella mutagenicity tests: IV. Results from the testing of 300 chemicals , 1988, Environmental and molecular mutagenesis.

[18]  E Zeiger,et al.  Identification of rodent carcinogens and noncarcinogens using genetic toxicity tests: premises, promises, and performance. , 1998, Regulatory toxicology and pharmacology : RTP.

[19]  L. Passador,et al.  Cell-to-Cell Communication in Bacteria , 2005 .

[20]  J. Casida,et al.  Glutathione activation of chloropicrin in the Salmonella mutagenicity test. , 1999, Mutation research.

[21]  H. Vainio,et al.  Mutascreen, an automated bacterial mutagenicity assay. , 1985, Mutation research.

[22]  R. Snyder,et al.  Comparison of the results of a modified miniscreen and the standard bacterial reverse mutation assays , 2000, Environmental and molecular mutagenesis.

[23]  Spiral Salmonella assay: Validation against the standard pour‐plate assay , 1996, Environmental and molecular mutagenesis.

[24]  L. Claxton,et al.  Development and validation of the spiral Salmonella assay: an automated approach to bacterial mutagenicity testing. , 1989, Mutation research.

[25]  J. Drake,et al.  An environmental mutagenesis test development programme. , 1980, IARC scientific publications.

[26]  E. Zeiger,et al.  Salmonella mutagenicity tests: V. Results from the testing of 311 chemicals , 1992, Environmental and molecular mutagenesis.

[27]  B. Burlinson,et al.  Use of the Miniscreen assay to screen novel compounds for bacterial mutagenicity in the pharmaceutical industry. , 1996, Mutagenesis.