Laboratory diagnosis, Treatment and Prevention of Streptococcus pneumoniae 4.83/5 (12)

Laboratory diagnosis, Treatment and Prevention of Streptococcus pneumoniae

Laboratory diagnosis of Streptococcus pneumoniae

Specimen: Sputum, blood, endotracheal aspirate, bronchoalveolar lavage, cerebrospinal fluid (CSF), pleural fluid, joint fluid, abscess fluid, bones, and other biopsy material.

Microscopy

  • Gram staining of sputum shows lancet shaped Gram-positive cocci in pairs.
  • Fresh emulsified sputum mixed with antiserum causes capsule swelling (the quellung reaction) for identification of pneumococci.
  • In acute pneumococcal otitis media, Gram stain of an aspirated fluid smear from middle ear is useful to demonstrate the bacteria.

Culture

  • Sputum or blood is plated on blood agar and incubated at 37° C in the presence of 5–10% carbon dioxide.
  • Gray colonies with alpha-hemolysis are observed after overnight incubation.
  • Diagnosis of pneumococcal meningitis is confirmed by CSF culture.

Identification of bacteria

  • Optochin sensitivity test
    • S. pneumoniae is identified by its sensitivity to optochin (ethylhydrocupreine dihydrochloride).
    • The isolate is streaked onto a blood agar plate and a disk saturated with optochin is placed in the middle of the inoculum.
    • A zone of inhibited bacterial growth is seen around the disk after overnight incubation.
  • Bile solubility test
    • It detects an autolytic enzyme, amidase, present in pneumococci, which breaks the bond between alanine and muramic acid of the peptidoglycan of the pneumococcal cell wall.
    • Isolates of S. pneumoniae are lysed rapidly when the autolysins are activated after exposure to bile.
    • Thus the organism can be identified by placing a drop of bile on an isolated colony.
  • Inulin fermentation test
    • It ferments inulin and hence differentiate it from other streptococci.

Animal inoculation

  • S. pneumoniae can be isolated from clinical specimens containing few pneumococci by intraperitoneal inoculation in mice.
  • Pneumococci are demonstrated in the peritoneal exudate and heart blood of the mice, which die 1–3 days after inoculation.

Antigen detection

  • Pneumococcal C polysaccharide is excreted in urine and can be detected using a commercially prepared immunoassay.
  • The CIEP is a useful test to detect pneumococcal capsular polysaccharide antigen in the CSF for diagnosis of meningitis, and in the blood or urine for diagnosis of bacteremia and pneumonia.
  • Latex agglutination test using the latex particles coated with anti-CRP antibody is employed to detect C reactive protein.
  • The CRP is used as a prognostic marker in acute cases of acute pneumococcal pneumonia, acute rheumatic fever, and other infectious diseases.

Antibody detection

  • The indirect hemagglutination, indirect fluorescent antibody test, and ELISA are used to demonstrate specific pneumococcal antibodies in invasive pneumococcal diseases.

Nucleic Acid–Based Tests

  • Nucleic acid probes and PCR assays are used for identification of S. pneumoniae isolates in culture.

Laboratory diagnosis, Treatment and Prevention of Streptococcus pneumoniae


Treatment of Streptococcus pneumoniae

  • Most pneumococci are susceptible to penicillin.
  • Other antibiotics, such as macrolides or selected fluoroquinolones with activity against pneumococci, are available for patients who are allergic to penicillin.
  • Penicillin-resistant pneumococci are being increasingly documented.
  • The mechanism of this resistance is an alteration of one or more of the bacterium’s penicillin-binding proteins (PBPs) rather than production of beta-lactamase.
  • Most resistant strains remain sensitive to third generation cephalosporins (such as cefotaxime or ceftriaxone), and all are still sensitive to vancomycin.
  • For serious pneumococcal infections, treatment with a combination of antibiotics is recommended. Vancomycin combined with ceftriazone is used commonly for empiric treatment, followed by monotherapy with an effective cephalosporin, fluoroquinolone or vancomycin.
  • Ceftriaxone can be used for meningitis caused by ceftriaxone-susceptible pneumococci.
  • Amoxicillin is the drug of choice for treatment of otitis media, sinusitis, and pneumonia caused by penicillin-resistant pneumococci with intermediate resistance.
  • Ceftriaxone is the drug of choice for non-CNS invasive pneumococcal diseases caused by penicillin- and ceftriaxone resistant pneumococci.
  • Vancomycin is used if the pneumococcus is resistant to ceftriaxone.

Prevention and control of Streptococcus pneumoniae

  • Pneumococcal vaccines: 23 valent pneumococcal polysaccharide vaccine (PPSV23) and polyvalent pneumococcal conjugate vaccine (PCV13) play an important role in prevention of pneumococcal diseases.
  • Personal hygiene is another important factor.

Laboratory diagnosis, Treatment and Prevention of Streptococcus pneumoniae

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