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Sci Rep. 2019 Jan 24;9(1):623. doi: 10.1038/s41598-018-36924-w.

Legionella SBT applied directly to respiratory samples as a rapid molecular epidemiological tool.

Author information

1
Infectious Diseases Unit, Fundació Institut d'Investigació Germans Trias i Pujol, Hospital Universitari Germans Trias i Pujol, Badalona, Spain. squero@igtp.cat.
2
Infectious Diseases Unit, Fundació Institut d'Investigació Germans Trias i Pujol, Hospital Universitari Germans Trias i Pujol, Badalona, Spain.
3
Infectious Diseases Unit, Fundació Institut d'Investigació Germans Trias i Pujol, Hospital Universitari Germans Trias i Pujol, Badalona, Spain. msabria.germantrias@gencat.cat.
4
CIBER de Enfermedades Respiratorias, CIBERES, Madrid, Spain. msabria.germantrias@gencat.cat.
5
Universitat Autònoma de Barcelona (UAB), Barcelona, Spain. msabria.germantrias@gencat.cat.
6
Vigilància Epidemiològica i Resposta a Emergències de Salut Pública, Agencia de Salut Pública de Catalunya, Barcelona, Spain.
7
CIBER de Enfermedades Respiratorias, CIBERES, Madrid, Spain.
8
Universitat Autònoma de Barcelona (UAB), Barcelona, Spain.
9
Infectious Diseases Unit, Fundació Institut d'Investigació Germans Trias i Pujol, Hospital Universitari Germans Trias i Pujol, Badalona, Spain. mgarcia@igtp.cat.
10
CIBER de Enfermedades Respiratorias, CIBERES, Madrid, Spain. mgarcia@igtp.cat.
11
Hospital Universitari Parc Taulí, Sabadell, Spain. mgarcia@igtp.cat.

Abstract

Legionnaires' disease (LD) is an atypical pneumonia caused by the inhalation of Legionella. The methods used for the diagnosis of LD are direct culture of respiratory samples and urinary antigen detection. However, the sensitivity of culture is low, and the urinary antigen test is specific only for L. pneumophila sg1. Moreover, as no isolates are obtained, epidemiological studies cannot be performed. The implementation of Nested-sequence-based typing (Nested-SBT) makes it possible to carry out epidemiological studies while also confirming LD, especially in cases caused by non-sg 1. Sixty-two respiratory samples from patients with Legionella clinically confirmed by positive urinary antigen tests were cultured and tested by Nested-SBT, following the European Study Group for Legionella Infections (ESGLI) protocol. Only 2/62 (3.2%) respiratory samples were culture-positive. Amplification and sequencing of Nested-SBT genes were successfully performed in 57/62 samples (91.9%). The seven target genes were characterised in 39/57 (68.4%) respiratory samples, and the complete sequence type (ST) was obtained. The mip gene was the most frequently amplified and sequenced. Nested-SBT is a useful method for epidemiological studies in culture-negative samples, achieving a 28.7-fold improvement over the results of culture studies and reducing the time needed to obtain molecular epidemiological results.

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