Combined fluorescent in situ hybridisation and immunolabelling of Bacteroides fragilis.
We have demonstrated that pure cultures of Bacteroides fragilis can be riboprobed with the oligoprobes BAC303 and EUB338, whilst simultaneously immunolabelled with either the mAb QUBF7, or polyclonal antiserum specific for a common antigen of B. fragilis. We were also able to distinguish between pure cultures of B. fragilis and Escherichia coli, by means of combined immunolabelling and riboprobing. The success of the combined technique is critically dependent on the size of the bacterial capsules, bacterial growth phase, antibody diluent and the length of the washing steps. The combined FISH and immunolabelling of bacteria has potential applications in studies of bacteria of medical and veterinary importance, as well as bacteria from other environments, as it yields information about both the identity and antigen expression of individual bacterial cells.
Department of Microbiology and Immunobiology, School of Clinical Medicine, The Queen's University of Belfast, Northern Ireland, UK.
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G
Ramage
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Patrick
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S
Houston
D000907:Antibodies, Bacterial; D015373:Bacterial Typing Techniques; D001441:Bacteroides fragilis / Q000145:classification*; D005455:Fluorescent Antibody Technique; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S / Q000302:isolation & purification; D015203:Reproducibility of Results
D000907:Antibodies, Bacterial; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S