III.
Chlamydiaceae
Description
∙ Non-motile, small (0.2-1.5 um), resemble
Gram-negative cell wall, obligate intracellular
parasites requiring living cells for growth
➔ Also known as an “Energy Parasite”
because it DOES NOT possess
Cytochromes
➔ Cytochrome is important for energy
production
➔ Requires living cells for growth
∙ Have tropism for columnar epithelial cells
∙ Species: C. trachomatis, Chlamydophila
psittaci, Chlamydophila pneumoniae
2 Morphological Forms
1. Reticulate Body (RF) - Replicative,
NON-INFECTIOUS FORM
−Intracellular, metabolically active form
–Diverts host cell’s functions to their own
metabolic needs
2. Elementary Body (EB) - INFECTIOUS
FORM
−Extracellular form; spherical; dense; resembles
Gram negative bacilli; has a rigid cell wall
−Infects host cell by requiring active
phagocytosis
−2 Components of outer membrane: MOMP
(Transmembrane protein), LPS Antigen
*MOMP - Major Outer Membrane Protein
➔ found only in Chlamydiaceae family
*LPS- Lipopolysaccharide
G.A.S.S
Note: Chlamydiaceae is the only family that
does not possess any cytochrome and cannot
synthesize ATP. Thus, they require a live cell as
their culture media.
I. Chlamydia trachomatis
∙ Major cause of Pelvic Inflammatory Disease
(PID) and Ocular Trachoma
∙ One of the major Sexually Transmitted
Pathogens
∙ Can travel through birth canal (can infect
infants)
∙ Associated with infertility and ectopic
pregnancy
∙ Natural Hosts: Humans
∙ Unique Feature: 10 stable plasmids (capable
of antibiotic resistance)
∙ 3 Biovars: Trachoma, Lymphogranuloma
Venereum (LGV), Mouse Pneumonitis
∙ 3 Serovars (Based on MOMP antigenic
differences)
1. A, B, Ba, C - Endemic Trachoma
2. L1, L2, L2a, L2b, L3 -
Lymphogranuloma Venereum
3. D-K, Da, Ia, Ja - PID, Urethritis,
Cervicitis, Epididymitis, Inclusion
Conjunctivitis
a. Clinical Infection
Trachoma
∙ Chronic inflammation of the conjunctiva
leading to blindness
∙ Cause distortion of the eyelids: eyelashes
become misdirected and turn in
∙ Transmission: fomites, hand-to-hand contact
with infected patient, by bite of flies
∙ Repeated exposure: Chronic Follicular
Keratoconjunctivitis, Conjunctival scarring,
Pannus
Lymphogranuloma Venereum
∙ Sexually transmitted disease
∙ Has a multisystem involvement
∙ Initially a small, painless ulcer or papule
(pimple-like) and develops as nodules
(buboes) after several weeks
∙ Intradermal skin test: Frei’s Test
The test was based on a positive
hypersensitivity to an intradermal
standardized antigen, lymphogranuloma
venereum, which indicated past or
present chlamydial infection.
Inclusion Conjunctivitis
∙ Usually infects infants upon delivery
∙ Copious, yellow discharge from the eye, an
inflamed and swollen conjunctiva, edematous
eyelids presence of large inclusion bodies in
host cell cytoplasm with an incubation period of
4-5 day
G.A.S.S
b. Laboratory Diagnosis
Specimens
∙ Swabs - Dacron or Rayon tipped swabs
∙ Urethra and Cervical secretions, Conjunctiva
discharges. NPS and Rectal swabs; Fallopian
Tube and Epididymis aspirates
Culture
∙ Reference method: Cell Culture
∙ McCoy cells, HeLa 229, Buffalo Green
Monkey Kidney Cells, Cycloheximide-treated
McCoy cells
∙ Prior to culture: Shell vial technique
(technique to concentrate the microorganism)
-Centrifuge specimen onto cell
monolayer, organisms are growing on a
coverslip in the bottom of the vial
∙After 48-72 hours: monolayers stained with
iodine or IF stains to examine presence of
inclusions
Others
∙ Cytologic Examination - cell scrapings from the
conjunctiva of newborns or persons with ocular
trachoma
∙ Enzyme Immunoassay - most used rapid
antigen assay
➔ Detects either outer membrane LPS
chlamydial antigen or the MOMP
∙
c. Other Tests
Antigen Detection and Nucleic Acid
Amplification
∙ More reliable in patients with symptomatic and
shedding large numbers of organisms
∙ DFA (Direct Fluorescent Antibody) assays
staining method using
Fluorescein-Isothiocyanate-conjugated
Monoclonal Antibodies: Identify in infected
cells
∙ Chemiluminescent DNA probe, ELISA, DFA:
Detect chlamydial antigen
∙ NAATs: Most sensitive method for detection
∙ LPS Antigen: Major antigen detected
∙ Specimen: Endocervical or Urethral Swabs
Serodiagnosis
∙ Primary antigen detected: LPS with
Ketodeoxyoctonate (EB) (from ppt) or Keto
Deoxy Octanoate (EB) (from google)
∙ If negative in serology tests: Reliably exclude
chlamydial infection
∙ COMPLEMENT FIXATION
*Detect family-reactive antibody; used
to diagnose LGV
*(+): Single point titer ≥ 1:64
(presumptive for LGV)
∙ MICROIMMUNOFLUORESCENCE ASSAY
(MICRO-IF)
*Used for type-specific antibodies for
C.trachomatis
*Used to diagnose
LGV(Lymphogranuloma venereum),
Trachoma, Inclusion conjunctivitis
* (+): IgM titer of 1:32
II. Chlamydophila psittaci (Former Chlamydia
psittaci)
G.A.S.S
● Causative agent of Psittacosis or Ornithosis
● Endemic pathogen of all bird species
(Parrots, Parakeets, Chickens, Ducks)
● Had outbreak among turkey-processing
workers and pigeon aficionados - can only be
cultivated under BSL-3 conditions
● Chlamydophila felis, Chlamydophila caviae,
Chlamydophila abortus - C. psittaci strains that
cause conjunctivitis, rhinitis on cats, guinea pig
conjunctivitis, and abortion among ruminants
● Acquisition - Inhalation of infected aerosols
from dried bird excreta or handling infected birds
● Diagnosis:
○ Complement Fixation - Titer ≥ 1:32
(Presumptive of Psittacosis)
○ Direct Microimmunofluorescence -
Sensitive Method
○ PCR-RFLP - Identify and distinguish
all 9 chlamydial spp.
III. Chlamydophila pneumoniae
∙ Formerly Chlamydia pneumoniae - TWAR
strain
➔ TWAR - Taiwan Acute Respiratory Agent
➔ TW-183 – Taiwanese child with
Trachoma
➔ AR-139 – Patient with Pharyngitis
∙ Human Pathogen; Transmission via aerosol
droplets via the respiratory route
∙ 3rd most common cause of infectious
respiratory disease
1. Streptococcus pneumoniae
2. Klebsiella pneumoniae (Ventilator
associated)
 3. Chlamydophila pneumoniae
4. Mycoplasma pneumoniae

∙ Associated with pneumonia, bronchitis,

pharyngitis, sinusitis
∙ Isolated from patients with Otitis media with
effusion

∙ Culture Media: HL or Hep-2 cell lines

∙ Method of choice: Micro-IF assay (Sensitive

and Specific)

Properties C. trachomatis C. psittaci C.

(Grp A) (Grp B) pneumoniae

Natural Humans Birds. Lower Humans

Hosts Animals

Elementary Round Round Pear-shaped

Body
(Infective
Stage)
Morphology

Inclusion Round, Vacuolar Variable, Round,

morphology Dense dense
& bodies **Halbertstadler- **Levinthal-C
Prowazek ole-Lillie**
bodies** (from
ppt)

**Halberstaedter
-Prowazek
bodies** (from
google)

Stains used Lugol’s Iodine Maquiavelo Giemsa

& Giemsa

Glycogen-co Present Absent Absent

ntaining
inclusions

Sulfonamide Susceptibility Resistant Resistant

(antibiotic)
susceptibility

Diseases Trachoma, LGV, Psittacosis Pneumonia,

Inclusion Pharyngitis,
conjunctivitis Bronchitis

Number of 20 10 1
serovars

G.A.S.S