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Detection of pneumolysin in sputum.

Abstract
Western blot detection of the species-specific pneumococcal product, pneumolysin (SPN), was shown to be almost as sensitive as PCR for the non-cultural detection of pneumococci in 27 Streptococcus pneumoniae culture-positive sputa from patients stated to have chest infections. Both techniques were considerably more sensitive than counter-current immuno-electrophoresis for pneumococcal capsular polysaccharide antigens (CPS-CIE) on the same specimens. Sensitivities for PCR, SPN-immunoblotting and CPS-CIE were 100%, 85% and 67%, respectively. In 11 S. pneumoniae culture-negative sputa taken from patients receiving antibiotics, but with proven recent pneumococcal infection, PCR and SPN-blot were positive in six (in two of which CPS-CIE was also positive), PCR alone was positive in one and SPN-blot alone was positive in one. In 11 S. pneumoniae culture-negative samples from patients not receiving antibiotics, all three tests were negative in eight, PCR was positive in three (in one of which CPS-CIE was also positive), but SPN-blot was negative in all 11. In 16 S. pneumoniae culture-negative samples from patients receiving antibiotics and with no known recent pneumococcal infections, one or more non-cultural test was positive in 11. Although further evaluation is required to assess the significance of pneumolysin detection in relation to carriage and infection and to devise a more suitable test format, these preliminary studies suggest that pneumolysin detection is a promising new approach to the non-cultural diagnosis of pneumococcal chest infection.
AuthorsJanice Wheeler, Roger Freeman, Michael Steward, Kirstine Henderson, Maureen J S Lee, Nigel H Piggott, Gary J A Eltringham, Angela Galloway
JournalJournal of medical microbiology (J Med Microbiol) Vol. 48 Issue 9 Pg. 863-866 (Sep 1999) ISSN: 0022-2615 [Print] England
PMID10482298 (Publication Type: Journal Article)
Chemical References
  • Antigens, Bacterial
  • Bacterial Proteins
  • Polysaccharides, Bacterial
  • Streptolysins
  • plY protein, Streptococcus pneumoniae
Topics
  • Antigens, Bacterial (analysis)
  • Bacterial Proteins
  • Blotting, Western (methods)
  • Counterimmunoelectrophoresis
  • Humans
  • Pneumococcal Infections (diagnosis, microbiology)
  • Polymerase Chain Reaction (methods)
  • Polysaccharides, Bacterial (analysis)
  • Respiratory Tract Infections (diagnosis, microbiology)
  • Sensitivity and Specificity
  • Sputum (chemistry, microbiology)
  • Streptococcus pneumoniae (isolation & purification, metabolism)
  • Streptolysins (analysis)

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