Use of heteroduplex analysis to classify legionellae on the basis of 5S rRNA gene sequences

Abstract

Seventeen different species of Legionella, 12 serogroups of Legionella pneumophila, and 2 Legionella-like amoebal pathogens (LLAP1 and Sarcobium lyticum) were examined by heteroduplex analysis of PCR products of the 5S rRNA gene. Eight different banding patterns were identified, indicating that heteroduplex analysis of this gene can be used to classify these bacteria according to base substitutions between species. This classification may have future applications in clinical and epidemiological studies.

Evaluation of Phenotypic and Genotypic Methods for SubtypingCampylobacter jejuniIsolates from Humans, Poultry, and Cattle

“…Further development of this new DGGE bacterial genotyping method will therefore involve selection of a smaller and less polymorphic DNA fragment. Furthermore, the heteroduplex analysis procedure has been shown in recent studies of Salmonella and Legionella typing to greatly enhance the precision and discriminatory power in nondenaturing assay systems (17,39).…”

The Use of Heteroduplex Analysis of Polymerase Chain Reaction Products to Support the Possible Transmission ofLegionella pneumophilaFrom a Malfunctioning Automobile Air Conditioner

“…Heteroduplex banding patterns, if present, were differentiated based on the number and relative positions of bands, compared with the homoduplex band, as previously described. 5 mens. Throat swab and serum specimens were positive on PCR for Legionella 5S rRNA, and urine was positive for L. pneumophila serogroup 1 antigen.…”

“…For heteroduplex analysis, amplified DNA (108 bp) from L. pneumophila serogroup 1 was used as a constant component, as described previously. 5 In a separate tube, 25 uL of the serum or water PCR product was mixed with 25 uL of the L. pneumophila constant PCR product and placed in a GeneAmp PCR System 2400 thermocycler (Perkin-Elmer). DNA strands were denatured at 95° C for 5 minutes and slowly cooled to 25° C by a 2-minute stay at every 5°.…”

“…3 ' 4 Surveys of physicians' and nurses' ballpoint pens have revealed bacterial contamination with methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), and gram-negative bacilli. 5,6 A recent report suggests that adhesive tape used to secure intravascular catheters may harbor bacteria and could lead to device-related infections. 7 Computer keyboards and faucets in an intensive care unit were found to be colonized with microorganisms, some of which were indistinguishable from patient isolates.…”

“…However, we report the possible transmission oi Legionella pneumophila serogroup 1 from a malfunctioning automobile air conditioning system's leaking water onto the floorboard of a car driven for a long distance by the patient. We have previously described the use of heteroduplex analysis to classify legionellae on the basis of the 5S rRNA gene sequences, 5 and this method was used in the current study to analyze Legionella polymerase chain reaction (PCR) products from an environmental and clinical source to establish a possible epidemiologic link.…”