A multileft evaluation of the Biotest legionella urinary antigen EIA

Harrison, Timothy G.; Uldum, S⊘en; Alexiou-Daniel, S.; Bangsborg, Jette; Bernander, Sverker; Drašar, Vladimír; Etienne, Jérôme; Helbig, Jörg; Lindsay, Diane; Lochman, I; Marques, Teresa; Ory, Fernando de; Tartakovskiĭ, I S; Wewalka, G.; Fehrenbach, F. J.

doi:10.1111/j.1469-0691.1998.tb00079.x

Cited by 39 publications

(28 citation statements)

References 11 publications

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“…Serologic analysis can be used only retrospectively and may be of limited value in immunocompromised patients. During recent years, screening kits for Legionella antigens in urine have become available (5). Alternatively, many groups have developed and evaluated assays to detect Legionella DNA in respiratory tract specimens and sometimes also in serum or urine (1, 6-8, 9-12, 14-16, 23).…”

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confidence: 99%

Development of Conventional and Real-Time PCR Assays for Detection of Legionella DNA in Respiratory Specimens

Rantakokko‐Jalava

Jalava²

2001

J Clin Microbiol

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The development and validation of a PCR assay based on the use of new 16S ribosomal DNA (rDNA)-targeted primers to detect Legionella DNA in respiratory specimens are described. The assay was originally developed as conventional PCR followed by electrophoretic detection and was then adapted to Lightcycler format with SYBR Green I detection and melting curve analysis. The 73 Legionella pneumophila strains tested were amplified with both applications. In addition, 21 and 23 out of 27 other Legionella strains were found positive by conventional and real-time PCR assays, respectively, including the clinically important species L. micdadei, L. bozemaniae, and L. dumoffii. Two DNA purification methods were compared using artificially seeded clinical specimens: a standard organic extraction method and a commercial kit based on adsorption of DNA to silica particles. The detection limit of the assay varied from 2 CFU to >200,000 CFU per ml of clinical specimen, depending on the background sample (i.e., pooled sputa or BAL fluids) and the DNA purification method, the silica method achieving lower detection limits. Analysis of 77 clinical samples (66 bronchoalveolar lavage fluid and 11 sputum samples) by conventional PCR yielded results that were consistent with Legionella culture results. The melting curve analysis in the Lightcycler system readily detected the specific amplification products. However, run-to-run variations in the measured melting temperatures required normalization against the standard sample in each run. The results obtained with the clinical specimens were similar to those obtained with conventional PCR, but more samples are required to determine whether the system can be applied to routine screening of samples for the presence of Legionella DNA.

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confidence: 99%

Development of Conventional and Real-Time PCR Assays for Detection of Legionella DNA in Respiratory Specimens

Rantakokko‐Jalava

Jalava²

2001

J Clin Microbiol

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“…The most common cause is Legionella pneumophila serogroup 1, however, just under half of the over 40 recognized species in the Legionellaceae family can cause Legionnaires' disease (2). A soluble antigen is present in the urine of patients with Legionnaires' disease due to serogroup 1 (3)(4)(5). A commercially available enzyme immunoassay is available that detects this antigen with a specificity of 100% and a sensitivity of 94.6% (3).…”

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Legionnaires' Disease - Results of a Multicenter Canadian Study

Marrie

Carolis

et al. 2003

Canadian Journal of Infectious Diseases

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BACKGROUND: There has never been a cross-Canada surveillance project to determine the rate ofLegionellaspecies as a cause of community-acquired pneumonia requiring hospitalization and to determine whether there are any regional differences in the rates of Legionnaires' disease in Canada. Anecdotally, Legionnaires' disease is thought to be uncommon in Western Canada.METHODS: From January, 1996 through to October 31, 1997, a prospective study of the etiology of community acquired pneumonia requiring admission to 15 tertiary care hospitals in eight Canadian provinces was conducted. A urine sample from each patient was tested forLegionella pneumophilaserogroup 1 antigen using a commercially available ELISA assay. A culture of sputum or other respiratory specimens for Legionellaceae was carried out at the discretion of the attending physician. Two hundred thirty-four patients had acute and 6-week convalescent serum samples tested for antibodies toL pneumophilaserogroups 1 through 6 using an ELISA method.RESULTS: 28 of the 850 patients (3.2%) had Legionnaires' disease; 18 of 823 (2.1%) were positive forL pneumophilaserogroup 1 by urinary antigen testing. The rate of Legionnaires' disease, based on urinary antigen, at the Halifax site was higher than that at the other sites (seven of 163 patients versus 11 of 660 [P=0.04]). Of the 28 cases of Legionnaires' disease identified using all methods, 11 of 277 patients (3.9%) were enrolled from Western provinces versus 17 of 573 patients (2.9%) from Eastern provinces (P=nonsignificant).CONCLUSIONS: Legionnaires' disease is just as common in Western as in Eastern Canada.L pneumophilaserogroup 1 may be more common in Halifax than at the other sites studied.

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“…Urinary antigen testing to detect Legionella antigen has proven the most powerful diagnostic method (4,5,15,16,18,22,24), and the commercially available tests have been widely used. However, the main drawback to the available urinary antigen tests is that they only detect the soluble antigen of L. pneumophila serogroup 1 but do not efficiently detect L. pneumophila non-serogroup 1 and other Legionella species (2,12). Moreover, concerns have been raised over the sensitivity of urinary antigen testing, as the prevalence of serogroup 1 decreases (27).…”

Section: Discussionmentioning

confidence: 99%

“…Two commercial EIA kits, Binax EIA (Binax, Portland, Maine) and Biotest EIA (Biotest AG, Dreieich, Germany), have been widely used since being marketed in 1996 and 1997, respectively, and a new Bartels EIA (Bartels, Inc., Trinity Biotech Company, Wicklow, Ireland) has been introduced recently. These EIAs have been reported to be sensitive and specific in many clinical studies (2,6,7,11,12,16,21). However, several authors pointed out that the available tests showed excellent sensitivity to L. pneumophila serogroup 1 antigen but variable sensitivity to L. pneumophila non-serogroup 1 and other Legionella species (2,6,7,12,16).…”

mentioning

confidence: 99%

“…These EIAs have been reported to be sensitive and specific in many clinical studies (2,6,7,11,12,16,21). However, several authors pointed out that the available tests showed excellent sensitivity to L. pneumophila serogroup 1 antigen but variable sensitivity to L. pneumophila non-serogroup 1 and other Legionella species (2,6,7,12,16). Although the L. pneumophila serogroup 1 is the predominant cause of legionellosis in most geographic areas, other pneumophila serogroups and other Legionella species are being recognized with increasing frequency, therefore questioning the broadspectrum utility of these tests (27).…”

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confidence: 99%

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Characterization of a Lipoprotein Common to Legionella Species as a Urinary Broad-Spectrum Antigen for Diagnosis of Legionnaires' Disease

Park¹,

Park²,

Chun

2003

J Clin Microbiol

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We have previously identified the Legionella 19-kDa peptidoglycan-associated lipoprotein (PAL) as a speciescommon immunodominant antigen. We describe here for the first time the excretion and detection of the PAL antigen in infected urine specimens, which is useful for the diagnosis of Legionnaires' disease. Rabbit anti-PAL immunoglobulin G (IgG) antibody was produced by immunization with the purified, recombinant PAL of Legionella pneumophila serogroup 1 and used in the PAL antigen capture enzyme-linked immunosorbent assay (ELISA) to detect urinary PAL antigen. A soluble-antigen capture ELISA using rabbit IgG antibodies against Legionella soluble antigens was prepared independently and used as a broad-spectrum standard test to detect soluble antigens of several Legionella species. Urine samples were obtained from guinea pigs experimentally infected with each of L. pneumophila serogroups 1, 3, and 6, and other Legionella species. The absorbance values of the PAL antigen ELISA highly correlated with those of the soluble-antigen ELISA in infected urine samples, with a correlation coefficient of 0.84 (P < 0.01). When applied to 17 infected urine samples and 67 negative controls from guinea pigs, the sensitivity and specificity of the PAL antigen capture ELISA were 88.2 and 95.5%, respectively. Compared to the commercial Biotest enzyme immunoassay, the PAL antigen ELISA was more efficient for detecting pneumophila non-serogroup 1 and nonpneumophila species. None of the 161 control human urine specimens obtained from healthy adults and patients with either non-Legionella pneumonia or urinary tract infections tested positive in the PAL antigen ELISA. The present study shows that the Legionella PAL is a very useful broad-spectrum antigen for urinary diagnostic testing. Moreover, since recombinant PAL antigen can be produced more efficiently than the soluble antigens, the development of a broad-spectrum diagnostic immunoassay based on the detection of the PAL antigen appears to be warranted.

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A multileft evaluation of the Biotest legionella urinary antigen EIA

Cited by 39 publications

References 11 publications

Development of Conventional and Real-Time PCR Assays for Detection of Legionella DNA in Respiratory Specimens

Development of Conventional and Real-Time PCR Assays for Detection of Legionella DNA in Respiratory Specimens

Legionnaires' Disease - Results of a Multicenter Canadian Study

Characterization of a Lipoprotein Common to Legionella Species as a Urinary Broad-Spectrum Antigen for Diagnosis of Legionnaires' Disease

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