API Industry General
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Transcript of API Industry General
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The Global Reference for Identification
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IDENTIFIKASI
Morfologi Pengamatan bentuk koloni dan mikroskopik
Genetik
Biokimia
Teknik biologi molekuler
Reaksi terhadap substrat biokimiawi, termasuk reaksi enzimatik
Serologi
Reaksi antigen dan antibodi spesifik
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Sistem identifikasi untuk bakteri dan jamur Dalam 1 strip terdapat 10 atau 20 substrat reaksi biokimiawi Tipe reaksi biokimiawi :
- Enzimatik - Assimilasi- Inhibisi
Total database =± 822 organisme (taksonomi berdasarkan pada International Journal of Systemic Bacteriology)
Microtube berisi substrat kering
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Tes enzimatik : menggunakan substrat kromogenik atau fluorogenik, seperti : aminopeptidase (Beta-alanine arylamidase), lipase, esterase, oxidase, ONPG, betaGAL, phosphatase
Tes asimilasi : menggunakan sumber karbon yang unik,
pendeteksian berdasarkan reaksi kekeruhan
seperti : karbohidrat, asam amino, asam organik
Tes inhibisi : menggunakan antibiotik atau substrat toksik, pendeteksian berdasarkan reaksi kekeruhan
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range
Jenis strip Gol. Bakteri/jamur InkubasiAPI 20EAPI 20E Gram negative Bacilli 24 jamAPI10E/10SAPI10E/10S Enterobacteria 24 jamRAPID 20ERAPID 20E Enterobacteria 4 jamAPI 20 NEAPI 20 NE Non Enterobacteria 24-48jamAPI StaphAPI Staph Staphylococci 24 jamAPI 20 StrepAPI 20 Strep Streptococci 4-24 jamAPI 20 C AuxAPI 20 C Aux Yeasts 24-48 jamAPI CandidaAPI Candida Yeasts 24 jamAPI 20AAPI 20A Anaerobes 24-48 jam API CampyAPI Campy Campylobacter 24 jamAPI CoryneAPI Coryne Corynebacteria 24 jamAPI NHAPI NH Neisseria/Haemophilus 2 jamAPI ListeriaAPI Listeria Listeria 24 jamAPI 50 CHBAPI 50 CHB Bacillus 24-48 jamAPI 50 CHLAPI 50 CHL Lactobacillus 24-48 jamAPI 50 CHEAPI 50 CHE Enterobacteria 24-48 jam
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Gallery
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Bagaimana cara memilih strip?
Lakukan tes orientasi :
• Morfologi koloni• Pewarnaan Gram • Tes katalase • Tes oksidase
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Kelengkapan
Mineral oilMineral oil
StripStripPSIpettes/ pipet steril
rak
Reagen tambahanReagen tambahanIncubation boxIncubation box
Medium suspensiMedium suspensi
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Prosedur Kerja
Suspensi
(McF)
Koloni
Inokulasi
HASILHASIL
Inkubasi
Pembacaan : Penambahan reagent
transfer
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Prinsip Inokulasi
GLU
Oil
ADH CIT
« anaerob fakultatif » « anaerobic » « aerobic »
Oil
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Pembacaan : Deteksi
Setelah proses inkubasi Reaksi dideteksi dengan cara :
perubahan warna secara spontan atau dengan penambahan reagent tambahan
Penyimpanan Penyimpanan :: 2 - 8°C2 - 8°C
VPB - NIT 2 - XYL - VP2 – HER
Dalam kondisi gelap :Dalam kondisi gelap : bungkus dengan aluminium foil
FB - FVB - ZYM B - NIN – PYZ – James - VP2
2 - 30°C2 - 30°C» VPA - VP1 - NIT 1 - Zn- BCP – TDA - ZYM A
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Pembacaan
Reaksi setelah inkubasi
Perubahan warna secara spontan
Perubahan warna dengan penambahan reagent
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Pembacaan : Deteksi
Gunakan tabel pembacaan (dalam package insert) dan isi dengan kode ( + ) atau ( - )
Kode tersebut ditransformasikan menjadi profil numerik dengan menggunakan prinsip pengkodean :
setiap tes pada strip dibagi menjadi 3 grup setiap tes bernilai : 1, 2 atau 4
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Kertas Kerja
7 angka profile 3604512
Hasil identifikasi dari software
Test tambahan
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PROSEDUR
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INTERPRETASI
Persentase reaksi positif untuk setiap reaksi dan takson
Taxon = base unit• Species• Biotype• Genus• Species group
bioMérieux Taxonomy = official taxonomy• International Journal of systematic bacteriology
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INTERPRETASI
CONTOH :
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INTERPRETASI
Identifikasi bakteri berdasarkan pada perbandingan profile terhadap database takson
Perbandingan memperhitungkan profile yang diperoleh dengan cara :
Mencari kesamaan dari berbagai takson yang berbeda dari database
% ID (Persen Identifikasi) Apakah isolat mikroorganisme tersebut berkaitan dengan suatu takson dalam database?Takson yang mana?
Mencari kesamaan dari typical profile dari setiap takson
Typicity index (T Index )Apakah isolat bakteri benar-benar terkait dengan takson tersebut ?
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Interpretasi
Taxon xA
B
C
Persen Identifikasi :% ID
• Nilai mendekati 100%• Kedekatan relatif ke berbagai takson dalam database
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Typicity index:T
C
the most typical profile
Taxon x
00
• Nilai mendekati 1 • Mendekati typical profile tertentu
Interpretasi
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Setiap hasil identifikasi selalu dilengkapi dengan
HARUS DILIHAT PERTAMA KALI: menunjukkan tingkat hasil identifikasi berdasarkan pada % of id, dan index T
excellent identification » % id > 99.9 et T > 0.75 « very good identification » % id > 99 et T > 0.5 « good identification » % id > 90 et T > 0.25 « acceptable Identification » % id > 80 et T > 0
Low discrimination : sistem tidak dapat membedakan 2 takson atau lebih Doubtful profile : terdapat tottaly test againts
INTERPRETASI
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Against tests: ?
Partially against tests
• Jika persentase positivitas < 25% dan test +• Jika persentase positivitas >75% dan test -
Totally against tests
• Jika persentase positivitas = 0% dan test +• Jika persentase positivitas 100% dan test –
INTERPRETASI
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Low discrimination : lakukan complementary tests Doubtful profile (test totally totally against) cek :
Kontaminasi
A typical organism
Technical errors (mineral oil, reagent, shelf-life …)
Ulang pengerjaan
INTERPRETASI
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APIweb
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Tes orientasi
Isolat muda
Isolat murni
Media yg standard
Kekeruhan
inokulum
Cara Inokulasi
Inkubasi
Rekomendasi Teknis
Penyimpanan reagent
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QUALITY CONTROLWhy ???
Memvalidasi proses transportasi strip API
Kualifikasi teknisi/pengguna
When ???
Setiap nomor lot baru
Setiap ada teknisi baru
How ???
Gunakan strain ATCC yang direkomendasikan oleh bioMérieux
Rekomendasi Teknis