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การตรวจวินิจฉัยเชื้อคอตีบ ทางห้องปฏิบัติการ PowerPoint PPT Presentation


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การตรวจวินิจฉัยเชื้อคอตีบ ทางห้องปฏิบัติการ. 9 พฤศจิกายน 2555. อารี ทัตติยพงศ์ Ph.D. กรมวิทยาศาสตร์การแพทย์ aree.t@dmsc.mail.go.th. โรคคอตีบทางห้องปฏิบัติการ. โรคคอตีบ.

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การตรวจวินิจฉัยเชื้อคอตีบ ทางห้องปฏิบัติการ

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9 2555

Ph.D.

aree.t@dmsc.mail.go.th


- C. diphtheriae diphtheria toxin

- C. diphtheriae diphtheria toxin


  • Corynbacterium diphteriae

  • Corynbacterium ulcerans

  • Corynbacterium pseudotuberculosis

Genus: Corynebacterium

Family: Corynebacteriaceae

Suborder:Corynebacteriaceae

Order: Actinomycetales

Phylum: Actinobacteria

Kingdom: Bacteria


C. diphtheriae

  • 1883 Klebs

  • 1884 Loffler

    (Klebs-Loeffler bacillus)

  • aerobic, facultative anaerobic bacteria

  • (club shape) V, L, Chinese character (palisade)

  • Catalase

  • Non-motile

  • Metachromatic granule


C. diphtheriae

  • 4 biotypes

    -var gravis

    -var intermedius

    -var mitis

    -var belfanti


C. diphtheriae

  • Disinfectant:

  • -1% sodium hypochlorite,

  • -70% ethanol,

  • glutaraldehyde,

  • formaldehyde,

  • iodines,

  • hydrogen peroxide,

  • peracetic acid,

  • quaternary ammonium compounds

Survive on dry inanimate surfaces from 7 days to 6 months


Colony BT

C. diphtheriae

C. diphtheriae


Sheep bloodagar Blood tellurite agar

24 hr.

Sheep bloodagar

Blood tellurite agar


Sheep bloodagar Blood tellurite agar

48 hr.

Blood tellurite agar

Sheep bloodagar


Tinsdale agar

C. diphtheriae

C. ulcerans

C. pseudotuberculosis

cysteinase

Corynebacterium diphtheriae on Tinsdale agar. The dark halo intensifies after prolonged cultivation. 72 hours, 37 C in an aerobic atmosphere


C. diphtheriae

  • Toxigenic strain vs non-toxigenic strain

  • Bacteriophage carry tox gene

  • Diphtheria toxin tox gene

  • C. diphtheriae strain infect Bacteriophage

  • Toxigenic strain can cause diphtheria


  • Pathogenesis

  • Toxin

    • tox gene ( bacteriophage)

    • exotoxin

    • A-B subunit

    • (pseudomembrane) >block airway

    • toxin myocarditis, neuritis etc


C. diphtheriae

  • 3. toxin Elek test


  • - 24 .

  • lab

  • lab

  • - lab 24

Throat swab

- swab (nylon, rayon, polyester) ()

- swab Amies media

antibiotic


Transport medium

  • overgrowth

Amies with charcoal

Amies without charcoal


Commercial Transport Medium

Amies without charcoal

Stuarttransport media



Throat swab

Sheep bloodagar Blood tellurite agar

18-24 hr

subculture Blood Agar

18-24 hr

(+ bacilli, pleomorphic, chinese letter) catalase

18-24 hr


Day 1

1. BT

2. Sheep BA

Throat swab

Incubate 18-24 hr


BT pure colony

Day 2

BT

colony subculture

Blood agar

Incubate 18-24 hr, 37OC

Blood agar -- -hemolysis Strep gr. A

.


catalase:+ oxidase: -

Day 3

Gram + bacilli,

pleomorphic , club shape

Chinese character

Gram staining

Incubate 18-24 hr, 37OC


Day 4

  • TSI

  • Motile

  • Bile esculin

  • Nitrate

  • Urease

  • Esculin

  • CTA-base

  • CTA-glucose

  • CTA-maltose

  • CTA-sucrose

  • CTA-mannitol

  • CTA-xylose

+

-

-

-

-

-

+

-

-

-

-

K/A


Streptococcus pyogenes(-hemolytic streptococcus gr. A)

-

- gram positive streptococci (cocci in chain)

- -hemolysis on sheep blood agar

-catalase negative, oxidase negative

-Sensitive Bacitracin

-Resistant Optochin

-PYR (L-pyrrolidonyl arylmidase) positive

-CAMP test -negative

-antiserum A-positive, B-negative


CAMP test negative

Gram positive cocci in chain

Bacitracin- sensitive

PYR- positive


Pure colony

  • C. amycolatum

  • C. diphtheriae

  • C. jeikeium

  • C. minutissimum

  • C. pseudodiphtheriae

  • C. striatum

  • C. ulcerans

  • C. urealyficum

Vitek 2 compact , automated bacterial identification, Drug sensitivity test


Biochemical patterns of C. diphtheriae


diphtheria toxin (Elek test)

Anti- toxin

Precipitation band

Immuno diffusion assay



C. diphtheriae : S. pyogenes =59:23 (2.6:1)


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