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Journal of Clinical Microbiology, May 2003, p. 2265, Vol. 41, No. 5
0095-1137/03/$08.00+0 DOI: 10.1128/JCM.41.5.2265.2003
Detection of Legionella pneumophila Serogroup 1 Antigen in Bronchoalveolar Lavage Fluid by an Immunochromatographic Assay

LETTER
Recently, an immunochromatographic assay for rapid qualitative
detection of
Legionella pneumophila serogroup 1 antigen in urine
specimens has become available (NOW
Legionella urinary antigen
test; Binax, Portland, Maine). We have previously shown that
this test is of clinical value in providing a rapid diagnosis
of Legionnaires' disease, especially in patients with severe
community-acquired pneumonia (CAP) in an outbreak setting (
5).
There are no reports on the applicability of the test to specimens
other than urine. Here we report results from a 32-year-old
human immunodeficiency virus (HIV)-infected man hospitalized
with CAP of the right upper lobe. Bronchoalveolar lavage (BAL)
was performed on the third day of admission because the patient
did not respond to empirical therapy with amoxicillin. Gram
staining of the BAL fluid revealed no apparent bacterial pathogens.
Ziehl-Neelsen staining showed no acid-fast bacteria. Giemsa
and toluidine blue stainings were negative for fungi. A urine
specimen obtained on the fifth day of admission tested positive
with the NOW
Legionella urinary antigen test. At that time,
culture of the BAL fluid yielded penicillin-resistant
Streptococcus pneumoniae and ß-lactamase-producing
Haemophilus influenzae.
Buffered charcoal yeast extract (BCYE) agar plates had been
incubated for less than 48 h and did not show growth of
Legionella spp. Subsequently, the pellet and supernatant of the stored
BAL fluid were tested with the NOW
Legionella urinary antigen
test, and both gave a positive result. To rule out cross-reactivity,
suspensions of the
S. pneumoniae and
H. influenzae isolates
of the patient were tested. Both suspensions tested negative.
The following day, an indirect immunofluorescent antibody (IFA)
test for
L. pneumophila (MONOFLUO
Legionella pneumophila IFA
test kit; Bio-Rad, Munich, Germany) was performed with the pellet
of the BAL fluid and small colonies growing at that time on
the BCYE agar plates. Both preparations showed brightly fluorescent
rods. The strain did not grow on BCYE in the absence of cysteine
and was identified as
L. pneumophila serogroup 1 by serogrouping
techniques (Dryspot
Legionella latex test; Oxoid, Hampshire,
United Kingdom). Antibacterial therapy was switched to levofloxacin
and cefotaxime. The patient was discharged from the hospital
in good condition after 14 days.
A preliminary evaluation of specificity of this application of the NOW Legionella urinary antigen test was conducted with BAL fluids from five pneumonia patients. L. pneumophila serogroup 1 antigen was not detected in pellets or supernatants of centrifuged BAL fluids, while culture yielded no Legionella spp. Conventional microbiological methods did reveal other microorganisms: e.g., H. influenzae, Serratia marcescens, Pseudomonas aeruginosa, Corynebacterium spp., coagulase-negative staphylococci, Candida spp., and Pneumocystis carinii.
The NOW Legionella urinary antigen test detected L. pneumophila serogroup 1 antigen in BAL fluid from an HIV-infected patient with culture-proven Legionnaires' disease. Application of the test to specimens other than urine might prove useful for rapid diagnosis of Legionnaires' disease. This may be especially true for anuric patients. End-stage renal disease is identified as risk factor for legionellosis, and acute renal failure is a known complication of Legionnaires' disease (1-3). The test might also prove useful for patients with extrapulmonary manifestations of L. pneumophila serogroup 1 infectione.g., pericarditis, which often presents without overt pneumonia (4). While our observation seems promising, additional clinical observations are needed to evaluate the sensitivity and specificity of the test with specimens other than urine.

REFERENCES
1 - England, A. C., III, and D. W. Fraser. 1981. Sporadic and epidemic nosocomial legionellosis in the United States. Epidemiologic features. Am. J. Med. 70:707-711.[CrossRef][Medline]
2 - Heule, H., F. W. Reutter, F. Gloor, and T. Giger. 1986. Renal involvement in Legionnaires' disease. Schweiz. Med. Wochenschr. 116:1736-1739.[Medline]
3 - Marston, B. J., H. B. Lipman, and R. F. Breiman. 1994. Surveillance for Legionnaires' disease. Risk factors for morbidity and mortality. Arch. Intern. Med. 154:2417-2422.[Abstract/Free Full Text]
4 - Stout, J. E., and V. L. Yu. 1997. Legionellosis. N. Engl. J. Med. 337:682-687.[Free Full Text]
5 - Wever, P. C., E. P. F. Yzerman, E. J. Kuijper, P. Speelman, and J. Dankert. 2000. Rapid diagnosis of Legionnaires' disease using an immunochromatographic assay for Legionella pneumophila serogroup 1 antigen in urine during an outbreak in The Netherlands. J. Clin. Microbiol. 38:2738-2739.[Abstract/Free Full Text]
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Peter C. Wever* Daan W. Notermans Igor I. Tulevski Jan K. M. Eeftinck Schattenkerk Menno D. de Jong
Departments of Medical Microbiology and Internal Medicine Academic Medical Center University of Amsterdam Amsterdam, The Netherlands
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* Phone: 31-20-5669111 Fax: 31-20-6979271 E-mail: p.c.wever{at}amc.uva.nl |
Journal of Clinical Microbiology, May 2003, p. 2265, Vol. 41, No. 5
0095-1137/03/$08.00+0 DOI: 10.1128/JCM.41.5.2265.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.