PCR-ELISA I: Application to simultaneous analysis of mixed bacterial samples composed of intestinal species.
Sixteen oligonucleotide identification probes, designed in this study or adapted from literature, were tested for a PCR-ELISA application to simultaneously detect under standardised conditions selected intestinal bacteria, lactobacilli and bifidobacteria. The level of specificity obtained with most of the probes fulfilled the set criteria. The lack of efficiency of PCR performed with the primers, proposed to be specific for the entire eubacteria domain, and compromises made in hybridisation conditions due to simultaneous usage of multiple probes reduced the sensitivity of the PCR-ELISA test. The method was, however, found to be suitable for detecting predominant members of the intestinal flora. Applicability of the PCR-ELISA test could be further widened using primers with a more restricted specificity in the PCR step, as was demonstrated for the detection of Bifidobacterium with genus-specific primers. Advantages of the PCR-ELISA method include convenient performance and the possibility to test rapidly large amounts of samples with a multitude of probes.
D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000032:analysis* / Q000145:classification / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007422:Intestines / Q000382:microbiology*; D007778:Lactobacillus / Q000235:genetics / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes