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Query: UMLS:C0023241 (Legionella)
 6,990 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Identification of six Legionella species, which we previously identified by serological test as Legionella bozemanii (L. bozemanii), was performed by DNA-DNA hybridization using a commercial DNA-DNA hybridization kit (Kobayashi Pharm. Co., Japan) introduced by Ezaki et al. All strains were identified as Legionella anisa (L. anisa), this being the first identification of L. anisa in Japan. Conventional laboratory tests were performed following the DNA-DNA hybridization. In this study the results of biochemical examination obtained, corresponded closely with those described in previous reports, but the oxidase reaction was very weak and varied according to the age of the culture, indicating the unreliability of this test in our case. All strains examined under long wave ultraviolet (UV) light (366 nm) revealed a blue-white fluorescence, the intensity of which ranged from strong to weak. Serological identifications were performed by both the slide agglutination test (SAT) and indirect immunofluorescent assay (IFA). SAT using commercially available antiserum (Denka Seiken., Japan) supposedly specific for L. bozemanii showed cross-reaction between L. bozemanii and L. anisa. Hyperimmune rabbit antisera prepared in this study for both L. bozemanii and L. anisa, from which cross-reactive antibodies were removed by the absorption of each antigen, reacted only with homologous antigens. IFA using a commercially available antiserum and hyperimmune rabbit antiserum previously described, gave positive reactions with each strain.
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PMID:Misleading serological identification of Legionella anisa as Legionella bozemanii. 140 75

Seventy-nine strains of legionella spp. and 13 non-legionella isolates were tested using a commercial latex agglutination kit. Cross-reactions were observed for all strains of Legionella pneumophila serogroup 12 examined between latex particles coated with rabbit antibodies raised against L. pneumophila serogroup 1 and those coated with rabbit antibodies to serogroups 2-14. Strains of L. pneumophila serogroup 1 did not cross-react with latex particles directed against L. pneumophila serogroups 2-14. This suggests that L. pneumophila serogroup 12 shares common surface antigens with L. pneumophila serogroup 1, and that conversely serogroup 1 isolates do not share major surface antigen determinants with serogroups 2-14. All other isolates tested gave expected results.
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PMID:Legionella pneumophila species identification using a commercial latex agglutination kit: a potential cross-reaction problem with serogroup 12. 148 81

Recent advance in molecular biology has enabled the specific and rapid diagnosis of the various infectious diseases. Though we commonly use the three major diagnostic procedure as isolation of the pathogen, direct detection of the pathogen and measurement of the immunological host reaction, DNA probe method would be the fourth major procedure in the clinical microbiology. The indication of the DNA probe method would be considered in the four cases as follows, 1. necessity of the special equipment to isolate the pathogen, 2. necessity of the long period to isolate the pathogen, 3. existence of the cross reaction among the pathogen and relative organisms in the immunological procedure, 4. existence of the difficulty to identify the species of the pathogen by the ordinary procedure. When we consider those indications, Legionnaires' disease might be one of the typical infectious disease to show the benefits of the DNA probe method in diagnosis. So far two types of DNA probe kits for Legionnaires' disease are available. One is the genus specific direct detection kit from the clinical specimens (Gen-probe), and the other is the microplate hybridization kit to identify each species of Legionella. The results of the evaluations of both kits showed the high specificity, rapidity and the clinical usefulness. In the next few years, various types of DNA probe kits might be newly developed and the contribution of those in the clinical microbiology would be much more than we expected.
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PMID:[Molecular diagnosis in respiratory infections]. 200 45

Specific DNA probes have been made for both M. pneumoniae and Legionella species. Dot blot methods have been used in research laboratories to test culture isolates of both organisms, and also to test animal tissues with a L. pneumophila-specific probe. Commercial kits are also available for direct specimen testing for these two organisms. The commercial kits are made by a single manufacturer, Gen-Probe, Inc. (San Diego, CA), and use a novel in-solution rapid hybridization assay, using 125I-labeled cDNA to rRNAs of the organisms. The Gen-Probe M. pneumoniae probe appears to be 80% to 100% sensitive, and 97% to 100% specific, based on analysis of two clinical studies using positive culture as the diagnostic criterion. The Gen-Probe legionella probe appears to be 33% to 71% sensitive (mean 57%), and 98.9% to 99.7% specific (mean 99.7%), based on analysis of four prospective clinical studies, using positive culture as the definition of disease, with a total sample size of 3,243 patients, 49 of which were culture-positive. Both Gen-Probe direct tests appear to be clinically useful, although the poor performance of the legionella test in one major university laboratory, and the expense of performing these tests, mandate that thorough evaluations be carried out in each laboratory anticipating using the test. Culture must always be performed for legionella whether or not the DNA probe test is used. It is likely that the use of the M. pneumoniae kit would greatly speed diagnosis, but whether this would alter medical practice or result in lower morbidity and health care costs is unknown.
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PMID:Use of DNA probes for the diagnosis of infections caused by Mycoplasma pneumoniae and legionellae--a review. 219 45

A commercial DNA probe kit designed to detect rRNA from legionellae was evaluated for its ability to correctly discriminate between legionellae and non-legionellae taken from culture plates. The probe kit, made by the Gen-Probe Corp. (San Diego, Calif.), was radiolabeled with 125I, and probe bacterial RNA hybridization, detected in a simple one-tube system hybridization assay, was quantitated with a gamma counter. A total of 156 Legionella sp. strains were tested, of which 125 were Legionella pneumophila and the remainder were strains from 21 other Legionella spp. A total of 106 gram-negative non-legionellae, isolated from human respiratory tract (81%) and other body site (19%) specimens, were also tested; 14 genera and 28 species were represented. The probe easily distinguished all of the legionellae from the non-legionellae. The average legionellae/non-legionellae hybridization ratio was 42:1, and the lowest ratio was 2:1; a minor modification in the procedure increased the lowest ratio to 5:1. In addition to correctly identifying all Legionella species, the probe was able to separate some of the various species of Legionella. L. pneumophila strains hybridized more completely to the probe than did the other Legionella spp.; L. wadsworthii and L. oakridgensis hybridized only about 25% of the probe relative to L. pneumophila. Some strains of phenotypically identified L. pneumophila had much lower hybridization to the probe than other members of the species and may represent a new Legionella species. The simplicity of the technique and specificity of the probe make it a good candidate for confirming the identity of legionellae in culture.
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PMID:Evaluation of the Gen-Probe DNA probe for the detection of legionellae in culture. 242 Aug 20

Confirmation of a culture as Legionella when it is unreactive with available serologic reagents involves tests that are impractical in most clinical laboratories. A nucleic acid probe that hybridizes only to members of the genus Legionella was recently prepared for marketing by Gen-Probe, Inc., San Diego, Calif. We tested 215 Legionella strains, representing 22 species, and 84 non-Legionella strains, representing 17 bacterial genera, with the Gen-Probe kit. All but four Legionella strains (L. bozemanii, less than 2% of total) and no heterologous strains gave positive test results. We conclude that the Legionella gene probe is a valuable addition to existing diagnostic tests for Legionella organisms.
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PMID:Evaluation of a commercial gene probe for identification of Legionella cultures. 242 99

We evaluated a 33-valent polyclonal indirect immunofluorescent-reagent kit (Merifluor-Legionella; Meridian Diagnostics Inc., Cincinnati, Ohio) made for the detection of Legionella species by testing bacterial isolates, seeded sputum, and negative sputum samples. Use of the reagent according to the directions of the manufacturer gave false-negative staining of homologous culture isolates due to a prozone phenomenon; this was solved by diluting test strain suspensions. After this change in testing protocol was made, the reagent gave bright fluorescent staining with 31 of the 33 Legionella strains with which it supposedly reacts. Strongly reacting Legionella strains included the type strains of L. pneumophila serogroups 1 to 10, L. longbeachae serogroups 1 and 2, and serogroup 1 of L. anisa, L. bozemanii, L. cherrii, L. dumoffii, L. gormanii, L. hackeliae, L. jamestowniensis, L. jordanis, L. maceachernii, L. micadedi, L. oakridgensis, L. rubrilucens, L. sainthelensi, L. spiritensis, L. steigerwaltii, and L. wadsworthii. Type strains of L. erythra and L. feeleii fluoresced only dimly with the reagent. Of 10 non-Legionella bacteria known to cross-stain with other polyvalent antisera, 5 also cross-reacted with the Merifluor reagent; these included 3 Bacteroides fragilis and 2 Pseudomonas fluorescens strains. The lower limit of detection of L. pneumophila serogroup 1 in seeded sputum was about 5 x 10(4) to 5 x 10(5) cells per ml. None of 21 randomly collected sputum specimens tested contained fluorescing legionellalike organisms, but 6 specimens did contain brightly fluorescing bacteria atypical in morphology for Legionella species. The Merifluor-Legionella kit appears to perform as well as other polyclonal immunofluorescent reagents used for detection of Legionella species. Because of the cross-reactions observed, which are common to all polyclonal reagents, utilization of this reagent for either bacterial identification or detection must be performed in combination with culture.
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PMID:Evaluation of the Merifluor-Legionella immunofluorescent reagent for identifying and detecting 21 Legionella species. 219 30

We used the Du Pont radioimmunoassay kit for soluble Legionella pneumophila serogroup 1 antigenuria (Du Pont Co., Wilmington, Del.) to test 422 urine samples from patients with and without Legionnaires disease (LD). The urine specimens were collected from 23 patients with culture-proven LD and from 346 patients without LD. L. pneumophila serogroup 1 was isolated from 14 patients with culture-proven LD, and other L. pneumophila serogroups or other Legionella species were isolated from 9 patients; 58 urine specimens were tested from these 23 patients. The non-LD group was composed of 75 bacteremic patients (35 gram-negative and 40 gram-positive bacteremias), 7 patients with candidemia, 48 patients with non-LD pneumonia, 90 patients with gram-negative bacteriuria (greater than 10(5) CFU/ml), 23 patients with gram-positive bacteriuria (greater than 10(5) CFU/ml), 14 patients with candiduria (greater than 10(5) CFU/ml), and 89 outpatients with negative urine cultures. All tests were performed in duplicate, including positive and negative controls. Sample results with values greater than or equal to 3.0 times those of the negative controls were considered positive for L. pneumophila serogroup 1 antigenuria. The average sample-to-negative ratios were 19.1 for the L. pneumophila serogroup 1 specimens, and 1.0 for both the non-serogroup 1 legionella group and the non-LD specimens. All but one of the patients who were culture positive for L. pneumophila serogroup 1 had at least one specimen positive for serogroup 1 antigenuria; none of the non-L. pneumophila serogroup 1 patients had a positive urine test. The test was highly specific (100%) and sensitive (93%) for the detection of L. pneumophila serogroup 1 antigenuria. Concentrations of urine by vacuum evaporation increased test sensitivity without apparently affecting specificity.
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PMID:Retrospective evaluation of the Du Pont radioimmunoassay kit for detection of Legionella pneumophila serogroup 1 antigenuria in humans. 318 24

A nonradioactive method is described that detects 10 to 100 legionellae in 1 ml of bronchoalveolar lavage fluid. DNA is purified by a proteinase K-phenol protocol or with a commercial DNA preparation kit and amplified by PCR with amplimers specific for the 16S rRNA gene of Legionella pneumophila. The upstream primer is 5' biotinylated. The amplification product is immobilized on streptavidin-coated microtiter plates. Because of the high binding capacity, no removal of nonincorporated biotin from the PCR product is required. After alkaline denaturation, the single-stranded PCR product is hybridized with a 5' digoxigenin-labeled probing oligomer. The amplification product is then detected by using peroxidase-labeled anti-digoxigenin antibodies in a luminescence or colorimetric reaction. The assay detects as few as 10 legionellae in 1-ml bronchoalveolar lavage fluid specimens. It is specific for medically relevant Legionella species, including Legionella pneumophila, L. bozemanii, and L. longbeachae. Of over 250 clinical specimens examined, 8 were positive for legionellae by both culture and the PCR assay. Six further specimens were culture negative but PCR positive for legionellae; of these, five specimens were from patients receiving high-dose erythromycin therapy for suspected or previously diagnosed legionella pneumonia. None of the remaining 240 specimens that were culture negative for legionellae yielded a positive PCR test, although a total of over 30 different bacterial species were cultured from these specimens. The PCR assay therefore appears to exhibit high sensitivity and specificity and thus could prove suitable for use in the routine microbiological diagnostic laboratory.
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PMID:Enzyme-linked immunoassay for detection of PCR-amplified DNA of legionellae in bronchoalveolar fluid. 754 66

Reclaimed water is an important resource for areas with inadequate water supplies. However, there have been few studies on the variety of microorganisms found in this type of water, since typically reclaimed water is examined only for the presence of coliform bacteria. Many microorganisms, including the legionellae, are known to be more resistant to chlorine than are coliform bacteria. Previously, we detected > 10(3) Legionella cells per ml in primary and secondary sewage effluents and observed no significant reduction in population numbers throughout the treatment process. In this study, we detected Legionella spp. in chlorinated effluent by using an EnviroAmp Legionella PCR kit and direct fluorescent antibody (DFA) staining. However, we were not able to isolate Legionella spp. from either natural or seeded reclaimed water samples. This suggests that the Legionella spp. detected by the PCR and DFA methods may be injured or viable but nonculturable after exposure to the high residual chlorine levels typically found in this type of water source. The numbers of coliform bacteria were low (< 2 cells per 100 ml) in most reclaimed water samples and were not correlated with the presence or absence of Legionella spp. We also collected air samples from above a secondary aeration basin and analyzed them by using the PCR, DFA, and plate culture methods. Legionella spp. were detected in the air obtained from above the secondary basin with all three methods. We concluded that the PCR was superior to the culture and DFA methods for detecting Legionella spp. in environmental water samples.
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PMID:Detection of Legionella species in reclaimed water and air with the EnviroAmp Legionella PCR kit and direct fluorescent antibody staining. 757 78


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