Identifikasi Bakteri

Oleh:Laila KaromahMoh. Mirza NuryadyAmin R. Setiawan

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Sistem Klasifikasi 3 DomainSistem Klasifikasi 3 Domain

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•Klasifikasi: mengelompokkan bakteri berdasarkan grup yang memiliki karakter sama

•Identifikasi: menyocokkan jenis bakteri tak dikenal dengan ciri-ciri bakteri lain yang telah dikenal sebelumnya

Fig 10.8KLASIFIKASI DAN IDENTIFIKASI BAKTERI

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Mengidentifikasi Bakteri?Mengidentifikasi Bakteri?

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Bergey’s Manual:Classifying and Identifying Prokaryotes 

Bergey’s Manual of Determinative BacteriologyProvides identification schemes for identifying bacteria and archaea

Morphology, differential staining, biochemical tests

Bergey’s Manual of Systematic BacteriologyProvides phylogenetic information on bacteria and archaea

Based on rRNA sequencing

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METODE IDENTIFIKASI BAKTERI

1. Microscopic morphology-Cell

2. Macroscopic morphology – colony appearance

3. Physiological / biochemical characteristics

4. Serological analysis

5. Genetic and molecular analysis

6. Phage typing

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MORFOLOGI SEL DAN KOLONI

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Morfologi SelMorfologi Sel

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Morfologi SelMorfologi Sel

Sumber: Hogg, 2005Sumber: Hogg, 2005

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MORFOLOGI SEL: Pengecatan Gram dan Pewarnaan Endospora

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Morfologi KoloniMorfologi Koloni

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BAGAN MORFOLOGI & PENGECATAN GRAM

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KARAKTER BIOKIMIA

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Hidrolisis dan Fermentasi Karbohidrat

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Hidrolisis Lemak

Bakteri ditumbuhkan pada media NA 10 ml + 0,4 ml

minyak nabati. Kemudian diinkubasi selama 2x24 jam,

suhu 37̊;C dituang CuSO4diamkan 5-10’dibuang

CuSO4

Positif bila daerah di sekitar koloni berwarna hijau

mengkilat

Contoh bakteri: Bacillus subtillis, E. coli, Pseudomonas,

Alcaligenes, Serratia, dan Micrococcus

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Hidrolisis Protein

Bakteri ditumbuhkan pada medium SMA(skim milk

agar)diinkubasi 3x24 jam, 37̊;Cpositif bila terdapat

zona bening di sekitar koloni

Contoh bakteri proteolitik: Bakteroides amyliphillus,

Clostridium sporogenes

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Menggunakan medium Nitrat cair

Reduksi Nitrat

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Uji Katalase (Reduksi H2O2)

Uji katalase ini digunakan untuk mengidentifikasi

bakteri pereduksi H2O2H2O + 1/2 O2 dengan enzim

katalase.

Uji ini positif bila terdapat gelembung O2 bebrapa saat

setelah penembahan H2O2 pada permukaan koloni

Contoh: Staphylococcus sp.

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Pembentukan H2S

Medium yang digunakan adalah TSIA(Triple Sugar Iron

Agar)

Hasil positif apabila media pertumbuhan berubah

warna menjadi hitam setelan inkubasi.

Contoh:Streptococcus sp., B. subtilis

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Numerical Rapid IdentificationNumerical Rapid Identification

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METODE GENETIK DAN METODE GENETIK DAN MOLEKULERMOLEKULER

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Serologymelibatkan serangkaian reaksi antara bakteri yang diamati dan antibody spesifik yang telah dikenal

Slide Agglutination

• Contoh:• Slide agglutination • ELISA

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Enzyme Linked Immunosorbent Assay - ELISAEnzyme Linked Immunosorbent Assay - ELISA

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Phage Typing

identifikasi jenis dan strain bakteri tak dikenal dengan menginfeksi bakteri tersebut dengan bakteriofaga tertentu

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Metode genetik/molekulerMetode genetik/molekuler• DNA fingerprinting:

menggunakan jumlah dan ukuran dari fragmen DNA yang diproduksi oleh RE untuk menentukan persamaan genetis

• Ribotyping: sekuensing rRNA

• PCR

Fig 10.14: Electrophoresis of RE digest of plasmid DNA

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Nucleic Acid Hybridization

Single strands of DNA or RNA, from related organisms will hydrogen-bond to form a double-stranded molecule; this bonding is called nucleic acid hybridization.

Examples of Applications:

• Southern blotting,

• DNA chips, and

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HUBUNGAN FILOGENI ORGANISME HUBUNGAN FILOGENI ORGANISME PROKARIOTPROKARIOT

Nucleic Acid Hybridization: DNA Chip

Fig 10.17