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Chapter 15 : Detection of Legionella spp. and Legionella pneumophila-Specific DNA in Respiratory Secretions by PCR-Enzyme-Linked Immunosorbent Assay and Comparison with Conventional Methods
Detection of Legionella spp. and Legionella pneumophila-Specific DNA in Respiratory Secretions by PCR-Enzyme-Linked Immunosorbent Assay and Comparison with Conventional Methods, Page 1 of 2< Previous page Next page > /docserver/preview/fulltext/10.1128/9781555815660/9781555813901_Chap15-1.gif /docserver/preview/fulltext/10.1128/9781555815660/9781555813901_Chap15-2.gif
This chapter talks about a study in which a PCR-enzyme-linked immunosorbent assay (ELISA) was devised for the detection of Legionella and Legionella pneumophila–specific DNA in human respiratory secretions. The PCR-ELISA incorporates a solid-phase probe hybridization step to verify the PCR reaction. Briefly, DNA was extracted from a total of 343 routine respiratory secretions using Nucleospin tissue columns. The PCR-ELISA was performed using reagents from the Roche Diagnostic PCR-ELISA (Dig Detection) kit Cat No. 1636111 following the manufacturers’ instructions. A total of 343 respiratory samples were tested, of which 21 patients were identified as cases of legionellosis by fulfilling one or more of the criteria for a definitive case. With PCR-based assays becoming more sensitive and specific, perhaps the distinction of a definitive case should incorporate more molecular techniques. The 16S RNA PCR-ELISA has the same sensitivity as the 5S RNA PCR/SB technique but can also identify L. pneumophila and is more sensitive than either direct fluorescent antibody (DFA) or culture. The lack of suitable samples, i.e., respiratory secretions, should not inhibit investigation, as Legionella specific DNA has been found in serum and urine.