Servicio de Información Comunitario sobre Investigación y Desarrollo - CORDIS

More universal and more rapid PCR methods to detect enterotoxin genes

The first purpose of this result is to permit a rapid detection of Bacillus cereus srains carrying enterotoxin genes (i.e. potentially able to cause diarrhoea). Three toxins coded by one gene (cytK) and by two operons of three genes each (hbl and nhe) have been showed to be able to cause diarrhoeal poisoning. New primers and a multiplex PCR method to amplify simultaneously three genes (cytK gene and one gene of the two operons hbl and nhe) have been developed.

The second purpose of this result is to permit a reliable PCR detection of Bacillus cereus strains carrying enterotoxin genes. Previously published primers for PCR amplification of nhe and hbl operon did not detect the operons in all Bacillus cereus strains, leading to false negative results. The sequence of enterotoxin genes from diverse Bacillus cereus strains was obtained to design more universal primers, able to detect the previously false negative strains. To increase reliabilityn, these primers were also designed to amplify in one-reaction long fragments covering the 3 genes of the hbl and nhe operons (the three genes are necessary for enterotoxic activity), whereas previously published primers detected only one gene for one operon.

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