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Laboratory diagnosis of legionnaires’ disease due to Legionella pneumophila serogroup 1: comparison of phenotypic and genotypic methods

¹Scottish Legionella Reference Laboratory, Department of Microbiology, Stobhill Hospital, Balornock Road, Glasgow G21 3UW, Scotland, UK ²Scottish Centre for Infection and Environmental Health, Clifton House, Clifton Place, Glasgow G3 7LN, Scotland, UK

Correspondence Diane S. J. Lindsay diane.lindsay{at}northglasgow.scot.nhs.uk

Received September 12, 2003
Accepted November 25, 2003

Laboratory results of 67 cases of legionnaires’ disease caused by Legionella pneumophila serogroup (Sg) 1 spanning a 6-year period were analysed by both phenotypic and genotypic methods. The methods compared were urinary antigen enzyme immunoassay (EIA), an immunofluorescent antibody (IFA) test, direct fluorescent antibody (DFA), culture and a 5S rRNA PCR with Southern blotting confirmation. Urine was available in 53 cases, of which 35 (66 %) were positive, with an antigen peak observed at 5–10 days after onset of disease symptoms. The IFA test was positive in 62 (92.5 %) cases, with 56 (90.3 %) cases producing a greater than fourfold rise in titre and 6 (9.7 %) giving presumptive high titres of 1 : 128. There were two antibody peaks, one at 10–15 days and another at >25 days after onset. In 23 cases where samples were available, DFA and culture were respectively positive in 5 (22 %) and 10 (48 %) cases. There was a peak in culture-positives 5–10 days after onset of disease. A Legionella-specific 5S rRNA PCR on patient serum was positive in 54 (80.5 %) cases, with a peak in PCR positivity at 6–10 days after disease onset. In 22 of the 67 cases, the full panel of diagnostic methods was available for comparison. The relative sensitivity and specificity of the urinary antigen EIA and the serum PCR was 100 %. The IFA gave relative sensitivity and specificity values of 93.8 and 95 %. DFA and culture, although 100 % specific, produced only low sensitivities, of 19 and 42.8 %, respectively. This study has shown that urinary antigen and serum PCR are valuable tests in the acute phase of disease, with excellent sensitivity and specificity values. At present, the Legionella species causing infection requires to be verified by IFA serology and/or culture, but this could become unnecessary as new antigen and L. pneumophila Sg 1-specific PCR tests become available.

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