title: Legionella pneumophila mip gene sequencing to investigate a cluster of pneumonia cases.
abstract: OBJECTIVE
  To use fluorescent antibody (FA) and PCR studies on fixed lung tissue to investigate whether Legionella pneumophila was the cause of pneumonia in a cluster of three haematology patients.
  
  
  METHODS
  Cut sections of paraffin blocks of lung tissue were examined by direct FA (DFA) using fluorescently labelled antibody to serogroup 1 and Pontiac strains of L. pneumophila. In addition, a single tube 'hanging drop' nested PCR targeting the mip gene of Legionella was performed on DNA extracted from the lung sections. Products were sequenced using dye terminator chemistry.
  
  
  RESULTS
  Numerous fluorescing bacteria were seen on staining with both antibodies in lung tissue from two of the patients. Identical L. pneumophila mip gene sequences were amplified from both DFA-positive lung sections. Two differing L. pneumophila mip sequences were obtained on three separate occasions from the tissue sections from the third patient negative by DFA. These sequences differed slightly from those obtained from the two DFA positive lung tissues.
  
  
  CONCLUSIONS
  There is good epidemiological evidence to link the first two cases who had been treated in the same ward prior to development of fever within two days of each other. The significance of results is controversial for the third patient.
authors.affiliation: Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Adelaide, South Australia, Australia. rod.ratcliff@imvs.sa.gov.au
authors.forename: Rodney M
authors.initials: RM
authors.lastname: Ratcliff
authors.forename: Monica A
authors.initials: MA
authors.lastname: Slavin
authors.forename: Norma
authors.initials: N
authors.lastname: Sangster
authors.forename: Robyn M
authors.initials: RM
authors.lastname: Doyle
authors.forename: John F
authors.initials: JF
authors.lastname: Seymour
authors.forename: Janice A
authors.initials: JA
authors.lastname: Lanser
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D001706:Biopsy; D005455:Fluorescent Antibody Technique; D005798:Genes, Bacterial / Q000235:genetics; D006801:Humans; D020104:Immunophilins / Q000235:genetics* / Q000378:metabolism; D016952:Legionella pneumophila / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D007877:Legionnaires' Disease / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D008168:Lung / Q000382:microbiology / Q000473:pathology; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D019696:Peptidylprolyl Isomerase; D011014:Pneumonia / Q000175:diagnosis* / Q000453:epidemiology / Q000209:etiology; D016133:Polymerase Chain Reaction
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D020104:Immunophilins; C512922:Mip protein, Legionella pneumophila; D019696:Peptidylprolyl Isomerase