A combined polymerase chain reaction-colour development hybridization assay in a microtitre format for the detection of Clostridium spp.
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.
Galindo
I
I
Gerencia Nacional de Biotecnología, Caracas, Venezuela.
Rangel-Aldao
R
R
Ramírez
J L
JL
D001431:Bacteriological Techniques; D001483:Base Sequence; D001709:Biotechnology; D003013:Clostridium / Q000235:genetics* / Q000302:isolation & purification*; D003116:Color; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D005069:Evaluation Studies as Topic; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D004269:DNA, Bacterial; D004275:DNA, Ribosomal