A gram stain is performed from the sputum of the infected patient.

The presence of neutrophils and

greater than ten gram-positive diplococci usually results in the diagnosis of Streptococcus pneumoniae.
For further conformation of this organism, it is streaked on blood agar. When on blood agar the
organism should exhibit alpha-hemolysis, which is characterized as a zone of green coloring around the
colonies of bacteria on the agar. This test isn’t always 100 percent conclusive because other
Streptococcus bacteria can cause alpha-hemolysis. Therefore, the streaked organisms must also exhibit
bile or optochin sensitivity to have greater assurance that this organism is in fact S. pneumoniae.

S. pneumoniae can be differentiated from the viridans streptococci, some of which are also alpha-

hemolytic, using an optochin test, as S. pneumoniae is optochin-sensitive. S. pneumoniae can also
be distinguished based on its sensitivity to lysis by bile, the so-called "bile solubility test".
The encapsulated, Gram-positive coccoid bacteria have a distinctive morphology on Gram
stain, lancet-shaped diplococci. They have a polysaccharide capsule that acts as a virulence
factor for the organism; more than 90 different serotypes are known, and these types differ
in virulence, prevalence, and extent of drug resistance.

Virulence Factors:

One virulence factor is a polysaccharide capsule that releases pneumococci from the host by preventing
phagocytosis. Another virulence factor is pnuemolysin which inhibits many things such as antibody
synthesis and lymphocyte proliferation. Inflammation is caused from the virulence factor of cell wall
components, which are thought to be the main cause of symptoms. There is also autolysin (enzyme)
which is a virulence factor that releases the cell wall products. (13

The bile (sodium deoxycholate) solubility test distinguishes S. pneumoniae from all other alphahemolytic

streptococci. S. pneumoniae is bile soluble whereas all other alpha-hemolytic

streptococci are bile resistant. Sodium deoxycholate (2% in water) will lyse the pneumococcal

cell wall