AGENT MORPHOLOGY
 Gram (+) coci in chain 1. Gram stain
 Catalase negative
 Facultative anaerobe
 Β haemolytic
Colonies on blood agar:
 Large and mucoid (1-2mm)
 Small zone of β hemolysis
(attributed to hemolysin)
 (+) Yellow, red, orange
pigment when incubated
anaerobically
 Small gram (-) bacteria
 Has inner and outer
membrane
 No peptidoglycan layer and
muramic acid
 Obligate intracellular
parasite (need host’s ATP
as energy source)
 Have DNA and RNA
Distinct forms:
 Elementary body
 Reticulate body
LAB DIAGNOSIS
- Presumptively identified by its ability to
2. Culture
1. Direct exam
- For chlamydial inclusions, antigens, or
nucleic acid
- Direct FA (fluorescent antibody) staining and
EIA (enzyme immunoassay) are used
- Tests are sensitive and can detect small
amounts of specific antigens or intact
organisms
2. Serologic diagnosis
- Done by complement fixation (CF) or
microimmuno-fluorescence (micro-IF)
techniques
3. Isolation
- Culture is through inoculation of infected
material into embryonated eggs or selected
tissue culture cell lines
OTHER INFO
hydrolyze sodium hippurate and by a
positive CAMP test
CAMP test
- Organism is inoculated perpendicular to the
streak of colonies of S.aureus and produces
an accentuated zone of complete hydrolysis
- CAMP factor (MW: 23.5kDa) is thermostable
and antigenic; acts in conjunction with
Staphyloccocal β lysine to complete RBC
membrane lysis
Distinct forms:
1.Elementary body
- metabolically inert, dense, round, & small
(0.2-0.4um)
- outer membrane has extensive disulfide
bond cross-linkages (confers stability for
limited extracellular existence)
2.Reticulate body
- intracellular metabolically active form
-divides by binary fission
-larger than EB (0.6-1um)
-synthesizes RNA, DNA, and proteins
  Rod-shaped organisms
 0.3-0.9um (width)
 2-3um (length)
 Motility is through flagella
Difficult to stain because:
- Not acid fast
- Doesn’t stain with
hematoxylin and eosin
- Will faintly stain gm (-) if
the safranin is left for an
extended period of time or
if a dilute solution of
carbofuchsin is used as
counterstain
Antigenic structure:
- 14 distinct serogroups
- All species have a single
common flagella antigen
Other agents: Streptococcus pneumoniae, H.influenzae type B, Streptococcus pyogenes, Staphylococcus aureus
1. Dieterle’s silver impregnation and direct
fluorescent antibody method
- Used to visualize organism in tissue
Biochemical and cultural characteristics:
- Best culture medium: charcoal yeast agar
(CYE) buffered with N-(2-acetamdo)-2-amino
ethane sulfonic acid (ACES)
- >5% CO2 inhibits growth
- Slowly growing, requires 3-5 days of
incubation
- Optimal temp: 37⁰C
- Doesn’t grow in temp >42⁰C
- Requires cysteine for growth on artificial
media
- Hydrolyzes starch, gelatin, and hippurate
- Catalase (+); weakly oxidase (+)
- Doesn’t fluoresce blue-white or red when
exposed to long wave UV light
- Produces brown pigment when grown in
media containing tyrosine
- Contains predominance of branched-chained
cellular fatty acids
 INFANTS
- There may be prodrome of URTI and diminished appetite
- Leads to abrupt onset of fever, restlessness, apprehension,
and respiratory distress
- Infants appear ill with respiratory distress manifested by
grunting, nasal flaring, retractions of supraclavicular and
inter/subcostal areas, tachypnea, tachycardia, air hunger,
and often cyanosis
- Some infants with bacterial pneumonia may have associated
GI disturbances (vomiting, diarrhea, anorexia, and abdominal
distention) secondary to paralytic ileus
- Rapid progression of symptoms is characteristic in the most
severe cases of bacterial pneumonia
ADULTS
- Community acquired pneumonia can vary from indolent to
fulminant in presentation and from mild to fatal in severity
- Patient is frequently febrile, tachycardic, may have chills
and/or sweats, cough (either non-productive or productive of
mucoid, purulent, or blood-tinged sputum)
- May experience pleuritic chest pain if pleura is involved
- Up to 20% may have GI symptoms
 Varies in shape (small
coccoidal to short,
branched filamentous
cells)
 Most distinctive feature:
bulbous enlargement w/
a differentiated tip
structure that can be seen
in young filamentous cells
grown in broth cultures
 Lacks peptidoglycan cell
wall
 Only protective layer: cell
membrane
1. Cold agglutinins Culture media
- Certain antigens present on human RBCs are - Media must be rich in cholesterol and
identical to antigens of the mycoplasma contain nucleic acids (purines and
membrane glycolipids pyrimidines)
- Antibodies (cold-agglutinins) to mycoplasma
antigens cross-react with human RBC
antigens and agglutinate the RBCs at 4⁰C
2. Complement fixation test
- Patient’s serum is mixed with glycolipid
antigens prepared from mycoplasma
- 4 fold rise in antibody titer between acute
and convalescent samples is diagnostic of
recent infection
3. Sputum culture
- “fried egg” appearance of colonies that
grown after 2-3 weeks
4. Mycoplasma DNA probe
- Sputum samples are mixed with a labelled
recombinant DNA sequence homologous to
that of the mycoplasma
- Recombinant probe will label mycoplasma
DNA if present