Simultaneous detection of Legionella species and L. anisa, L. bozemanii, L. longbeachae and L. micdadei using conserved primers and multiple probes in a multiplex real-time PCR assay☆

Details

• Alternative Title:
  Diagn Microbiol Infect Dis
• Personal Author:
  Cross, Kristen E. ; Mercante, Jeffrey W. ; Benitez, Alvaro J. ; Brown, Ellen W. ; Diaz, Maureen H. ; Winchell, Jonas M.
• Description:
  Legionnaires' disease is a severe respiratory disease that is estimated to cause between 8,000 and 18,000 hospitalizations each year, though the exact burden is unknown due to under-utilization of diagnostic testing. Although Legionella pneumophila is the most common species detected in clinical cases (80-90%), other species have also been reported to cause disease. However, little is known about Legionnaires' disease caused by these non-pneumophila species. We designed a multiplex real-time PCR assay for detection of all Legionella spp. and simultaneous specific identification of four clinically-relevant Legionella species, L. anisa, L. bozemanii, L. longbeachae, and L. micdadei, using 5'-hydrolysis probe real-time PCR. The analytical sensitivity for detection of nucleic acid from each target species was ≤50fg per reaction. We demonstrated the utility of this assay in spiked human sputum specimens. This assay could serve as a tool for understanding the scope and impact of non-pneumophila Legionella species in human disease.
• Subjects:
  DNA Primers Humans Legionella Legionnaires' Disease Multiplex Polymerase Chain Reaction Oligonucleotide Probes Real-Time Polymerase Chain Reaction Sensitivity And Specificity
• Source:
  Diagn Microbiol Infect Dis. 85(3):295-301
• Pubmed ID:
  27107536
• Pubmed Central ID:
  PMC5505572
• Document Type:
  Journal Article
• Funding:
  CC999999/Intramural CDC HHSUnited States/
• Volume:
  85
• Issue:
  3
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:5bd1c9e73612c89bcba84cb2c97baa04adefdf98867793ddbe38b2e8c1aa246c
• Download URL:
  https://stacks.cdc.gov/view/cdc/46698/cdc_46698_DS1.pdf
• File Type:
  [PDF - 797.97 KB ]