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Slide 001
METODE KIMIA
TAHAP DASAR ORIENTASI
PEMERIKSAAN HBA1C METODE
IMMUNO TURBIDIMETRIPresentan : Tisha Patricia
Pembimbing : Dian Ariningrum, dr., M.Kes, Sp.PK,
PK 001
Program Pendidikan Dokter Spesialis Patologi Klinik
Fakultas Kedokteran Universitas Sebelas Maret
Surakarta
20161
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TAHAP DASAR: ORIENTASINO
.
METODE JUDUL TANGGAL
PRESENTASI
PEMBIMBING
1.
2.
3.
4.
5.
6.
Sekresi
Imuno
Serologi
Mikrobiologi
Sampling
Hematologi
Kimia-1
Analisis Cairan
Sinovial
Pemeriksaan
Toxoplasma gondii
Pada Toksoplasmosis
Serebral dengan
Metode
ELISA
Kultur Kuantitatif Ulkus
Kaki Diabetik
Preparasi Sampel:
Pipetasi
Aggregometry
Pemeriksaan HbA1c
Metode
Immunoturbidimetri
13 - 10 - 2015
27 - 11 - 2015
14 - 12 – 2015
30 – 12 - 2015
2 - 2 – 2016
21-4-2016
Yuwono Hadisuparto, dr., Sp. PK
Tonang Ardyanto, dr., Phd, Sp.PK
B. Rina Sidharta, dr., Sp.PK-K
Amiroh Kurniati, dr., M. Kes., Sp.PK
Amiroh Kurniati, dr., M. Kes., Sp.PK
Dian Ariningrum, dr., M. Kes, Sp. PK
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Learning Objective
1. Menjelaskan metode pemeriksaan HbA1c
2. Menjelaskan prinsip pemeriksaan
turbidimetri dan immunoturbidimetri
3. Menjelaskan faktor pra-analitik, analitik
dan post analitik pemeriksaan HbA1c
metode immunoturbidimetri
Slide 002
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BAB I PENDAHULUAN
439 juta 2030
30% tidak menyadaripenyakitnya
25% sudah menderita komplikasimikrovaskular
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HbA1c
Skrining
Diagnosis
Monitoring
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http://anthonycerami.org/site/hba1c/
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Weykamp C, 2013. HbA1c: analytical and
clinical aspects,
www.annlabmed.org 9
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Harefa E, 2010. Peran HbA1c dalam
skrining dan diagnosis diabetes melitus.
Informasi Laboratorium Klinik Prodia;3:1-3.
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Prinsip turbidimetri
http://intranet.tdmu.edu.ua/data/kafedra/intern
al/pharma_2/classes_stud/en/pharm/prov_pha
rm/ptn/analytical%20chemistry/2%20course/1
9%20Optical%20methods%20of%20the%20an
alysis.htm
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PEMERIKSAAN HBA1C
METODE
IMMUNOTURBIDIMETRI
BAB II
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• Penentuan kuantitatif HbA1c dalam
sampel whole blood dengan antikoagulan
EDTA.
Tujuan
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PRA ANALITIK
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Prinsip Pemeriksaan
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a. Automatic/semiautomatic biochemistry
analyzer
b. Haemolysis Reagent 1 mL
c. R1: 300 µl
d. R2: 100 µl
Alat dan Bahan
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Komposisi reagen
a. Haemolysis
Reagent
– H2O
– Stabilizers
b. Reagen R1
– Latex 0,1%
– Glycine buffer, pH
3,0, 15 mmol/L
c. Reagen R2
– Goat anti-mouse IgG
polyclonal antibody 0,8mg/dl
– Mouse anti-human HbA1c
monoclonal antibody
0.05mg/ml
– Glycine buffer, pH 3.0,
60mmol/L
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a.Pipet 20 µl dan 1 ml
b.Test tube
c.HbA1c calibrator set dan kontrol
Alat dan bahan yang dibutuhkan tetapi
tidak disediakan
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• Pemeriksaan darah tanpa persiapan
khusus
• Sampel: darah vena dengan
antikoagulan EDTA
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1 ml Hemolysis reagent + 20 µl whole blood diaduk
Tunggu 5’ lisis
Penundaan pemeriksaan: hemolisatdisimpan pada suhu 2-8oC, 10 hari
Preparasi Hemolisat
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• Stabilitas reagen: pada suhu 2-
8oC
Setelah dibuka reagen stabil hingga
1 bulan
• Kalibrasi: 5 level kalibrasi
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ANALITIK
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Prosedur pemeriksaan
www.biowaydx.com/_pdf/Reagent/R032K11%20HbA1c.pdf28
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http://pubs.rsc.org/services/images/RS
Cpubs.ePlatform.Service.FreeContent.I
mageService.svc/ImageService/Articlei
mage/2015/AY/c4ay02904f/c4ay02904f-
f2_hi-res.gif29
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PASCA ANALITIK
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Konsentrasi HbA1c disajikan langsung
dalam bentuk % HbA1c menggunakan ΔOD
untuk melihat kesesuaiannya pada kurva
kaliberasi.
Hasil
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Non diabetes < 6%
Diabetes terkontrol HbA1c < 7%.
Setiap laboratorium diharapkan menetapkan
kisaran nilai HbA1c sendiri
Nilai yang diharapkan
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• Hasil pemeriksaan mungkin tidak
inkonsisten pada pasien dengan
–adiksi terhadap opiate
–Alcoholism
–pasien yang menggunakan
aspirin dosis tinggi
Keterbatasan
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• Linearity:
2%-16%
• Presisi
–Within Run: CV≤5%
–Run-to-run ≥5%
Karakteristik Performa
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Interferensi
• Tidak ada interferensi terdeteksi untuk
kadar Bilirubin 0,5g/L (50mg/dL),
• asam askorbat 0,5g/L (50mg/dL),
• Trigliserida 20 g/L (2000 mg/dL),
• Hb Karbamasil (7,5 mmol/L), dan
• Hb asetilasi (5,0 mmol/L)
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1. Prinsip pemeriksaan HbA1c metode
immunoturbidimetri: ikatan antibodi
dengan glukosa dan antara asam amino-
4 dengan 10 N-terminal rantai β.
Pemeriksaan tidak dipengaruhi oleh HbE,
HbD maupun carbamylated Hb.
BAB III
SIMPULAN
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2. Pemeriksaan HbA1c metode
immunoturbidimetri sudah dapat
diotomatisasi, dapat dipakai untuk
laboratorium dengan jumlah sampel yang
lebih kecil karena diperiksa dengan
analiser untuk kimia yang lain.
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DAFTAR PUSTAKA
ADA 2010. Diagnosis and classification of Diabetes Mellitus. Diabetes Care, 2012; 35(supplement 1): S64-S71.
American Diabetes Association (ADA), 2009. Standards of Medical Care in Diabetes—. Diabetes Care; 32.
Anik Widijanti dan Resti Anggun Pertiwi, 2015. Perbandingan HbA1c Metode Immunoassay dan IE-HPLC pada Faal Ginjal Normal.
Medika. Tahun ke XLI, No. 5, 286–291
Anonim 1, Bioway Chemistry Reagent Series Glycated Hemoglobin (HbA1c) Detection Kit,
www.biowaydx.com/_pdf/Reagent/R032K11%20HbA1c.pdf
Anonim 2, Prime Evolution, Semi-automatic biochemistry analyzer / bench-top / with immunoturbidimetry dosing. http://www.
medicalexpo.com/prod/bpc-biosed/product-67898-436281.html
Anonim 3, 2013. Tina-quant® HbA1c Gen. 3 Turbidimetric inhibition immunoassay (TINIA) for the in vitro determination of hemoglobin
A1c in whole blood or hemolysate. http://www.cobas.com/content/dam/cobas_com/pdf/product/Tina-quant-HbA1c-Gen-3/Tina-
Quant%20HbA1c%20Fact%20Sheet.pdf
Anonim 4, 2014. Peran pemeriksaan HbA1c pada diabetes mellitus. http://www.abclab.co.id/?p=1209
Gallagher EJ, Roith D, Bloomgarden Z, 2009. Review of hemoglobin A1c in the management of diabetes. Journal of Diabetes; 1 : 9–17.
Gough S, Manley S, Stratton I, 2010. HbA1c in diabetes case studies using IFCC units. Wiley Blackwell; 2-61.
Harefa E, 2010. Peran HbA1c dalam skrining dan diagnosis diabetes melitus. Informasi Laboratorium Klinik Prodia;3:1-3.
Inzucchi SE, Bergenstal RM, Buse JB, et al, 2012. Management of Hyperglycemia in Type 2 Diabetes: A Patient-Centered Approach,
American Diabetic Association (ADA). Diabetes Care Journal ;10: 1- 4.
Khan MI, Weinstock RS, 2007, Carbohydrates. In Mc Pherson RA, Pincus MR, 21th eds. Henry’s Clinical Diagnosis and Management by
Laboratory Methods. China: Saunders Elsivier, 185-199.
Little RR, Rohlfi ng CL, 2009. HbA1c standardization: Background, progress and current issues. Lab Med. 2009;40:368-73.
Monnier L, Collete C, 2009. Target for glycemic control concentrating on glucose. Diabetes Care 2009;32:199-203.
NGSP, 2004. IFCC Standardization of HbA1c, http://www.ngsp.org/ifccngsp.asp
Perkumpulan Endokrinologi Indonesia (PERKENI), 2011. Konsensus Pengelolaan dan Pencegahan Diabetes Melitus tipe 2 di Indonesia.,
Perkumpulan Endokrinologi Indonesia, p:1-72.
Saudek CD, Derr RL, Kalyani RR. Assessing glycemia in diabetes using self-monitoring blood glucose and hemoglobin A1c. JAMA
2006;295:1688-97.
Sofronescu AG, Williams LM, Andrews DM, ZhuY. Unexpected Hemoglobin A1c Results. Clin Chem 2011;57(2):153-157.
Sri Rahayu Paputungan dan Harsinen Sanusi, 2014. Peranan Pemeriksaan Hemoglobin A1c pada Pengelolaan Diabetes Melitus. CDK-220/
vol. 41 no. 9: 650-655
WHO 2011. Use of Glycated Hemoglobin (HbA1c) in the Diagnosis of Diabetes Mellitus. 38
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TERIMA
KASIH
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