300412 - Intro Lab - Mikrobiologi - Dr. Harvi
-
Upload
meutia-wilda -
Category
Documents
-
view
97 -
download
3
Transcript of 300412 - Intro Lab - Mikrobiologi - Dr. Harvi
![Page 1: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/1.jpg)
LOG
Diagnosis of Laboratory Diagnosis of Laboratory MicrobiologyMicrobiology
Harvi Puspa WardaniDept. of Microbiology
Medical SchoolUNISBA
2011/2012
![Page 2: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/2.jpg)
ww
w.t
hem
eg
allery
.com
LEARNING OBJECTIVE
Memahami prinsip identifikasi bakteri
ReferensiCappucino, Sherman : Microbiology a
Laboratory ManualJawetz : Medical Microbiology
Company LogoCompany Logo
![Page 3: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/3.jpg)
Bacterial Infection (Secondary Infection)(sign / symptoms)(Etiologic agent)(Site of Infection)
CLINICAL SPECIMENS(kind of specimens)
Enrichment medium Direct Smear (Gram Staining)
Blood agar Mac Conkey agar(Measures of colony, Reaction on blood agar) (Morphology of colony, Reaction on M Conkey agar)
Gram staining Gram Staining Gram Positive Gram Negative
Catalase Lactose fermenter Non-Lactose fermenter
(E. coli (Salmonella Klebsiella Shigella)
Enterobacter)
Catalase (-) Catalase (+) (Streptococcus sp.) (Staphylococcus sp.)Strept. Pneumoniae KIA (Optochin +, Inulin +) S. aureus (Coagulase +) MIUStrept. Viridans S. epidermidis (Novobioc +) Citrate Strept. Hemolyticus S. saprophyticus
![Page 4: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/4.jpg)
ww
w.t
hem
eg
allery
.com
Catalase -
Streptococcus
β-hemolytic α-hemolytic γ-hemolytic
pyogenes agalactiae Enterococcus pneumonia viridans
Bacitracin Bacitracin optochin s optocin r
Sensitive resistant inulin +
Company LogoCompany Logo
![Page 5: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/5.jpg)
ww
w.t
hem
eg
allery
.com
DIRECT SMEAR (PEWARNAAN GRAM)
Agar dapat terlihat pada preparat dengan pewarnaan Gram konsentrasi minimun bakteri dari cairan tidak pekat : 104-105 organisme/ml
Kegunaan :
- Menentukan spesimen untuk kultur adekuat/tidak
- Membuat etiologi presumptiveSputum,CSF,material dari luka atau abses
pewarnaan Gram secara langsungSpesimen yang tidak cocok dilakukan pewarnaan
Gram secara langsung : apus tenggorok, feses, darah
Company LogoCompany Logo
![Page 6: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/6.jpg)
ww
w.t
hem
eg
allery
.com
Blood agar
Nilai : reaksi hemolysis, ukuran koloni, warna,elevasi
Hemolysis : suatu reaksi bakteri di sekitar koloni yang dapat terlihat pada medium
Terbagi menjadi : α hemolysis, β hemolysis, γ hemolysis
Company LogoCompany Logo
![Page 7: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/7.jpg)
ww
w.t
hem
eg
allery
.com
α-hemolysis
Gambaran warna hijau disekitar koloni
Lisis RBC inkomplit Hb menjadi
MethemoglobinCo : Streptococcus
pneumoniae and streptococcus viridans.
Company LogoCompany Logo
![Page 8: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/8.jpg)
ww
w.t
hem
eg
allery
.com
hemolysis
Gambaran bening di sekitar koloni
Lisis RBC dengan destruksi komplit dan penggunaan Hb oleh mikroorganisme
Terbentuk krn adanya 2 tipe beta hemolysin : Streptolysin O dan Streptolysin S
Contoh : Streptococcus pyogenes,
Company LogoCompany Logo
![Page 9: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/9.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Ukuran koloni
Besar, sedang, kecil, pintpointBakteri Gram-positive secara umum koloni lebih
kecil dibandingkan Gram negatif. Streptococci khas membentuk koloni pinpointStaphylococci koloni lebih besar (pinhead)
WARNA
Keemasan : S.aureus
![Page 10: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/10.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Elevasi
Tinggi, convex, flat, umbilicate (tengahnya ada lekukan), umbonate (tengahnya timbul).
Co : S. pneumoniae : koloni umbilicate.
![Page 11: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/11.jpg)
ww
w.t
hem
eg
allery
.com
MacConkey Agar
Medium selective dan media differensial
Tdp crystal violet menghambat pertumbuhan bakteri Gram +
Laktosa, bile salt, dan pH indikator neutral red membedakan bakteri bdsrkan kemampuan memfermentasikan laktosa
- Lactose fermenter koloni meras
- Nonlactose fermenter tidak berwarna
Company LogoCompany Logo
![Page 12: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/12.jpg)
ww
w.t
hem
eg
allery
.com
Perbedaan Dinding Bakteri Gram Negatif dan Positif
Company LogoCompany Logo
![Page 13: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/13.jpg)
ww
w.t
hem
eg
allery
.com
Pewarnaan Gram
Prinsip : Reagen kimia :
- Zat warna dasar (Primary stain)
- Penguat (Mordant)
- Decolorizing agent
- Counterstain
Company LogoCompany Logo
![Page 14: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/14.jpg)
ww
w.t
hem
eg
allery
.com
Gram Stain (cont’)
Crystal violet sbg primary stain semua sel berwarna ungu
Iodine meningkatkan afinitas sel thd zat warna. CV-I membentuk insoluble complexmeningkatkan intensitas warna purple black
Ethyl Alcohol 95% sbg decolorizing agent.Fungsi: protein-dehydrating agent dan lipid solvent.
Company LogoCompany Logo
![Page 15: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/15.jpg)
ww
w.t
hem
eg
allery
.com
Gram Stain (cont’)
Gram (-) : lipid pada membran sel akan larut shg porositas dinding meningkat. Komplek CV-I mudah dihilangkan. Sel menjadi tidak terwarnai
Gram (+) : lapisan peptidoglikan tebal berperan dalam lebih kuatnya retensi kompleks CV-I, pori-pori dinding lebih kecil akibat efek dehidrasi alkohol.
Company LogoCompany Logo
![Page 16: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/16.jpg)
ww
w.t
hem
eg
allery
.com
Gram Stain (cont’)
Safranin sbg counterstain menyebabkan sel menjadi merah pada sel yang tidak terwarnai
Hasil : Gram + : ungu Gram - : merah
Company LogoCompany Logo
![Page 17: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/17.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Tes Katalase
Prinsip: Catalase
2H2O2 2H2O + O2
(+) : bubblesReagen : 3% hydrogen
peroxide Staphylococcus:
Catalase+ Streptococcus : Catalase
–
![Page 18: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/18.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Tes Koagulase
Enzim bekerja di sel inang mengubah fibrinogen menjadi fibrin.
S.aureus : Coagulase (+)S.epidermidis dan S.saprophyticus : coagulase (-)
![Page 19: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/19.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Fermentasi Inulin
S.pneumoniae dapat memfermentasikan inulin sedangkan alpha hemolytikus streptococci lainnya inulin-nonfermenter
![Page 20: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/20.jpg)
ww
w.t
hem
eg
allery
.com
Tes Optochin
ethylhydrocupreine hydrochloride, a quinine derivative
Streptococcus pneumoniae sensitif terhadap optochin
Pertumbuhan bakteri yg sensitif optochin akan dihambat disekitar optochin disc, sementara pertumbuhan bakteri yg resisten optochin tidak akan dihambat
Company LogoCompany Logo
![Page 21: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/21.jpg)
ww
w.t
hem
eg
allery
.com
TSIA
Terdapat glukosa (butt), laktosa dan sukrosa (slant)
iron utk mendeteksi produksi hydrogen sulfide hitam
Kuning asam di medium (A) , tidak ada perubahan warna alkaline (K)
Produksi gas gelembung pada butt
Company LogoCompany Logo
![Page 22: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/22.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
![Page 23: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/23.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
KIA
Mengandung laktosa ( pada slant) dan glukosa (pada butt)
Jika terjadi fermentasi KH :Juga mengandung sodium thiosulfat dan ferrous
sulfate indikator pembentukan H2S
Na2S2O3 + bacterial acids H2S + Fe+2 FeS black precipitate
Artinya : H2S (+)
![Page 24: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/24.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
KIA (cont’)
Escherichia, Klebsiella, Enterobacter acid slant, acid butt, No H2S
Shigella alkaline slant, acid butt, no H2S
Most Salmonella alkaline slant, acid butt, H2S produced
![Page 25: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/25.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Tes Indole
Tujuan : menentukan kemampuan m.o. mendegradasikan asam amino tryptophan.
Prinsip:Tryptophanase
Tryptophan Indole + Pyruvic acid + Ammonia
Kovac’s reagent + indole red violet compound
![Page 26: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/26.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Tes Urease
Urease Urea + 2H2O CO2 + H2O + 2NH3
Ammonia alkaline environment :
menyebabkan phenol red menjadi pink tua
![Page 27: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/27.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Tes Citrate
Tujuan: menentukan kemampuan m.o. memfermentasikan sitrat sebagai sumber karbon.
Prinsip:
Citrase
Citrate Oxalocetic acid + acetic acid pyruvic acid + CO2
CO2 + 2Na+ + H2O Na2CO3 alkaline pH perubahan warna dari hijau ke biru
![Page 28: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/28.jpg)
ww
w.t
hem
eg
allery
.com
Laboratory test for differentiation of Staphylococcal sp.
Test S. aureus S.epidermidis S.saprophyticus
MSA :-Growth-Fermentation
+
+
+
-
+
-
Coagulase + - -
DNase + - -
Hemolysis beta - -
Novobiocin test
S S R
Pigmentation Golden yellow
white white
![Page 29: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/29.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo
Enterobacter Escherichia Klebsiella Most
Salmonella
Shigella
TSI slant A A (K) A K K
Butt AG AG AG A;G + A
Motility + +/- - + -
Indol - + +/- - +/-
Urease - - +/- - -
Citrate + - + +/- -
![Page 30: 300412 - Intro Lab - Mikrobiologi - Dr. Harvi](https://reader034.fdocuments.in/reader034/viewer/2022042615/55cf98d4550346d03399ec11/html5/thumbnails/30.jpg)
ww
w.t
hem
eg
allery
.com
Company LogoCompany Logo