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การตรวจวินิจฉัยเชื้อคอตีบ ทางห้องปฏิบัติการ PowerPoint PPT Presentation


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การตรวจวินิจฉัยเชื้อคอตีบ ทางห้องปฏิบัติการ. 9 พฤศจิกายน 2555. อารี ทัตติยพงศ์ Ph.D. กรมวิทยาศาสตร์การแพทย์ [email protected] [email protected]การ. โรคคอตีบ.

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การตรวจวินิจฉัยเชื้อคอตีบ ทางห้องปฏิบัติการ

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3672892

9 2555

Ph.D.

[email protected]


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- C. diphtheriae diphtheria toxin

- C. diphtheriae diphtheria toxin


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  • Corynbacterium diphteriae

  • Corynbacterium ulcerans

  • Corynbacterium pseudotuberculosis

Genus: Corynebacterium

Family: Corynebacteriaceae

Suborder:Corynebacteriaceae

Order: Actinomycetales

Phylum: Actinobacteria

Kingdom: Bacteria


C diphtheriae

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 diphtheriae1

C. diphtheriae

  • 4 biotypes

    -var gravis

    -var intermedius

    -var mitis

    -var belfanti


C diphtheriae2

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


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Colony BT

C. diphtheriae

C. diphtheriae


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Sheep bloodagar Blood tellurite agar

24 hr.

Sheep bloodagar

Blood tellurite agar


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Sheep bloodagar Blood tellurite agar

48 hr.

Blood tellurite agar

Sheep bloodagar


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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 diphtheriae3

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


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  • Pathogenesis

  • Toxin

    • tox gene ( bacteriophage)

    • exotoxin

    • A-B subunit

    • (pseudomembrane) >block airway

    • toxin myocarditis, neuritis etc


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C. diphtheriae

  • 3. toxin Elek test


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  • - 24 .

  • lab

  • lab

  • - lab 24

Throat swab

- swab (nylon, rayon, polyester) ()

- swab Amies media

antibiotic


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Transport medium

  • overgrowth

Amies with charcoal

Amies without charcoal


Commercial transport medium

Commercial Transport Medium

Amies without charcoal

Stuarttransport media


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Throat swab

Sheep bloodagar Blood tellurite agar

18-24 hr

subculture Blood Agar

18-24 hr

(+ bacilli, pleomorphic, chinese letter) catalase

18-24 hr


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Day 1

1. BT

2. Sheep BA

Throat swab

Incubate 18-24 hr


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BT pure colony

Day 2

BT

colony subculture

Blood agar

Incubate 18-24 hr, 37OC

Blood agar -- -hemolysis Strep gr. A

.


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catalase:+ oxidase: -

Day 3

Gram + bacilli,

pleomorphic , club shape

Chinese character

Gram staining

Incubate 18-24 hr, 37OC


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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

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


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CAMP test negative

Gram positive cocci in chain

Bacitracin- sensitive

PYR- positive


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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


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Biochemical patterns of C. diphtheriae


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diphtheria toxin (Elek test)

Anti- toxin

Precipitation band

Immuno diffusion assay


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


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