Applied Genetics and Regulation

Applied Microbiology and Biotechnology

, Volume 39, Issue 4, pp 553-557

A combined polymerase chain reaction-colour development hybridization assay in a microtitre format for the detection of Clostridium spp.

  • Iván GalindoAffiliated withGerencia Nacional de Biotecnología, Cervecería Polar, C.A.Laboratorio de Genética Molecular, Facultad de Ciencias, Universidad Central de Venezuela
  • , Rafael Rangel-AldaoAffiliated withGerencia Nacional de Biotecnología, Cervecería Polar, C.A.Universidad Simón Bolívar
  • , José Luis RamírezAffiliated withLaboratorio de Genética Molecular, Facultad de Ciencias, Universidad Central de Venezuela

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Abstract

We have developed a rapid and sensitive assay for the detection of clostridial cells or spores in liquid food samples. The method recognizes Clostridium 16S (small ribosomal subunit RNA) rDNA (ribosomal DNA) sequences by a polymerase chain reaction-digoxigenin-labelling protocol, coupled to a capture oligonucleotide immobilized on a microtitre plate. The positive results are revealed by means of a colour reaction. In 6 h of non-intensive labour, we can detect as few as two to five clostridial cells or spores in experimentally contaminated soft drinks.