CORYNEBACTERIUM DIPHTERIAE

OVERVIEW (Morphology and Identification)

- Gram (+) rods aerob

- Small (0,5-1 mikrometer in diameter and several micrometers long)
Bentuknya rods dengan irregular swellings di ujungnya  “club
shaped appearance”, Chinese letter pattern (acute angle with each
other)
- Non motile, non spore forming
- 4 biotypes: gravis, mitis, intermedius, belfanti
- Pleomorphism (kemampuan suatu bakteri untuk mengubah ukuran
dan bentuk in response to environmental conditions)
- Ada yang toxigenic dan nontoxigenic
C. diphteriae tidak perlu menjadi toxigenic untuk membentuk infeksi lokal, namun yang
nontoxigenic tidak punya efek lokal/sistemik  bisa bikin penyakit tapi milder respiratory
symptomsnya dan tidak membentuk pseudomembrane.

PATHOGENESIS

- Diphteriae itu principally a result of the action of the toxin formed by

the organism rather than invasion of the organism.
- C. Diphteriae infections terjadi di respiratory tract, wounds, or skin,
spread by droplets or contact dengan susceptible individuals  tumbuh
di mukus membrane atau skin abrasions, lalu yang toxigenic akan
menghasilkan toxin (exotoxin).
- Faktor-faktor yang mempengaruhi keluarnya toxin: konsentrasi Fe,
osmotic pressure, amino acid concentration, pH, availability
PATHOLOGY

- Tempat infeksi: respiratory tract, wound, or skin.

RESPIRATORY TRACT

- Exotoxin diserap mucous membranes  destruction of epithelium  superficial inflammatory

response  necrotic epithelium terbenam dalam exuding fibrins dan RBC, WBC  membentuk
grayish pseudomembrane (secara umum over the tonsils, pharynx, larynx).
- Any attempt untuk remove the pseudomembrane akan menyebabkan si kapiler terkekspos dan
robek  bleeding
- Regional lymph node di leher membesar atau marked edema di entire neck.
- Diphteriae bacili dalam membrane terus menerus menghasilkan toxin  distant toxic damage 
parenchymatous degeneration, fatty infiltration, necrosis di otot jantung, liver, kidney, adrenals,
kadang diikuti gross hemorrhage. Bisa juga nerve damage, menyebabkan paralysis di soft palate,
eye muscles/extremities.

WOUND OR SKIN DIPHTERIAE

- Membrane dapat terbentuk di infected wound yang fails to heal. Absorpsi toxinnya sedikit, dan
efek sistemik jarang terjadi.

Tidak actively invade deep tissues dan practically never enters the bloodstream namun rare bisa
menyebabkan endocarditis.

DIAGNOSTIC LABORATORY TESTS

- Prinsip: fungsinya untuk konfirmasi clinical impression dan merupakan epidemiologic significance
Sehingga specific treatment tidak boleh didelay cuma gara-gara hasil lab kalau misalnya clinical
picturenya udah specific diphteriae.
- Specimen: dacron swabs dari throat, nose, wounds, or lesions (Lebih baik taken from the
membrane). Kemudian diletakkan pada semisolid transport medium seperti Amies.
- Lalu specimennya inoculated (dikenalkan pada 3 medium):
1. Loeffler’s serum slope
Mengandung horse serum, meat infusion, dextrose
Fungsi: untuk menumbuhkan diphteriae bacili untuk confirm the diagnosis
Setelah incubation 6 jam atau semalaman  diphteriae like morphology
2. Tellurite plate (cystine-tellurite agar/ modified Tinsdale medium)
Mengandung potassium tellurite: inhibitor untuk other respiratory
flora
Dalam 36-48 jam akan tumbuh tampak black colonies with halos
karena tellurite reduced intracellularly
3. Blood Agar
Fungsi untuk rule out hemolytic
streptococci
Semuanya incubated aerobically at 370
- C. Diphterae from clinical material or culture ketika distain:
a. Alkaline methylene blue/Loeffler’s stain

b. Gram staining

c. Neisser’s staining

d. Albert’s staining

Green colored rod shaped bacteria yang arranged at angle to each other (chinese letter),
black metachromatic granules at poles

Albert dan Neisser’s stain: untuk stain the metachromatic granules  granules yang irregularly
distributed, kebanyakan di pores

- Virulence test untuk mengecek toxigenicity:

a. Modified Elek immunoprecipitation method described by the World Health Organization
Diphtheria Reference Unit.A filter paper disk containing antitoxin (10 IU/disk) is placed on an
agar plate. The cultures to be tested (at least 10 colonies should be chosen) for toxigenicity
are spot inoculated 7–9 mm away from the disk. After 48 hours of incubation, the antitoxin
 diffusing from the paper disk has precipitated the toxin diffusing from toxigenic cultures and
has resulted in precipitin bands between the disk and the bacterial growth.
b. Polymerase chain reaction (PCR)–based methods have been described for detection of the
diphtheria toxin gene (tox). PCR assays for tox can also be used directly on patient specimens
before culture results are available. A positive culture result confirms a positive PCR assay. A
negative culture result after antibiotic therapy along with a positive PCR assay result
suggests that the patient probably has diphtheria.
c. Enzyme-linked immunosorbent assays can be used to detect diphtheria toxin from clinical C
diphtheriae isolates.
d. An immunochromatographic strip assay allows detection of diphtheria toxin in a matter of
hours. This assay is highly sensitive. The latter two assays are not widely available.