seminar idi barabai 9 mei 2015 revisi (dr. yanti sp.pk).pptx
TRANSCRIPT
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LABORATORY FINDING IN DENGUE VIRUS
INFECTION
dr. Faizah Yunianti Sp.PKRSUD. H.Damanhuri - Mitra Laboratorium
Barabai
Struktur virus dengue
Virus dengue
DEN 3
DEN 2 DEN 4
2
DEN 1
Famili Flavivirida
e
ArbovirusØ 30 nm
2
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penyakit flu, demam tifoid, demam chikungunya, leptospirosis, malaria
Tidak khas
Sulit diagnosisTerutama pada masa
awal demam (3-4 hari)
Manifestasi klinis
Beragam : asimtomatik, undifferentiated febrile illness, demam dengue, demam berdarah dengue (DBD) ,perembesan plasmasyok hipovolemik (SSD),kematian
Infeksi virus dengue
Manifestasi laboratoris
Trombositopenia ringan sampai dengan berat,
berbagai kombinasi kehadiran NS1, IgG dan IgM dan variasi kadar sitokin.
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Peran pemeriksaa
n laboratoriu
m
4
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Detection of the virus / virus isolation (culture)PCRDetection of antidengue antibody or antigen
(serology)LeukopeniaLymphocytosisThrombocytopeniaRaised of hematokrit
Laboratory Diagnosis
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Leukopenia,Thrombocytopenia, Raised of hematokrit
lymphocytosis
Puskesmassederhana
bilik hitung, reangensia mikroskop
Rumah Sakit DL Otomatikbilik hitung, reangensia mikroskop
Hapusan darah
True/ False
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Nama : kasus 1 Umur : YPARAMETER HASIL NILAI NORMAL
Hemoglobin (Hb) 12,1 12,0 – 17,4g/dL
Eritrosit (RBC) 3,9 4,0 - 5,5 X 1012 / L
Hematokrit (Ht) 49,0 36,0 – 52,0 %
MCV 81 76 - 96 fL
MVH 29,0 27,0 – 32,0 pg
MCHC 33,0 31,0 – 38,0 g/dL
RDW CV 14,3, 0,0 – 16,0%
Lekosit (WBC) 3,3 5,0 – 10,0 X 109/ L
Eosinofil 1,0 0,0 – 2,0 %
Basofil 0,0 0,0 – 1,0%
Neutrofil 34 50 – 70 %
Limfosit 65 20 - 40%
Monosit 6 2 – 8 %
Trombosit (Plt) 57 150 – 450 X 109/ L
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Nama : kasus 2 Umur : YPARAMETER HASIL NILAI NORMAL
Hemoglobin (Hb) 12,1 12,0 – 17,4g/dL
Eritrosit (RBC) 3,9 4,0 - 5,5 X 1012 / L
Hematokrit (Ht) 36 36,0 – 52,0 %
MCV 81 76 - 96 fL
MVH 29,0 27,0 – 32,0 pg
MCHC 33,0 31,0 – 38,0 g/dL
RDW CV 14,3, 0,0 – 16,0%
Lekosit (WBC) 3,3 5,0 – 10,0 X 109/ L
Eosinofil 1,0 0,0 – 2,0 %
Basofil 0,0 0,0 – 1,0%
Neutrofil 77 50 – 70 %
Limfosit 32 20 - 40%
Monosit 6 2 – 8 %
Trombosit (Plt) 57 150 – 450 X 109/ L
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Nama : kasus 3 Umur : YPARAMETER HASIL NILAI NORMAL
Hemoglobin (Hb) 7,3 12,0 – 17,4g/dL
Eritrosit (RBC) 3,9 4,0 - 5,5 X 1012 / L
Hematokrit (Ht) 36 36,0 – 52,0 %
MCV 54 76 - 96 fL
MVH 29,0 27,0 – 32,0 pg
MCHC 33,0 31,0 – 38,0 g/dL
RDW CV 14,3, 0,0 – 16,0%
Lekosit (WBC) 4.7 5,0 – 10,0 X 109/ L
Eosinofil 1,0 0,0 – 2,0 %
Basofil 0,0 0,0 – 1,0%
Neutrofil 54 50 – 70 %
Limfosit 45 20 - 40%
Monosit 6 2 – 8 %
Trombosit (Plt) 165 150 – 450 X 109/ L
Virus IsolationDefinitive test for dengue infectionLab equipped with tissue culture facilitiesUseful only at early phase of illness , blood
collected before day 5 of illness ( before the formation of Neutralizing antibodies)
During febrile illnessVirus can be isolated from serum, plasma and
leucocytesPost mortem specimens
Expensive and time consuming (2 weeks to complete
Polymerase Chain Reaction (PCR)
Diagnosis of dengue infection in the early phase (< 5 days of illness)
Sensitivity 100% in the first 5 days of illness, reduced to 70% by day 6
Determine dengue serotype
Polymerase Chain Reaction (PCR)Limitation
Available in certain facilities and trained personal
Test is expensiveSpecimens requires special storage
temperatures and short transportation , time between collection and extraction
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Deteksi AntigenNS1 (Non struktural Antigen 1)
Deteksi AntibodiIg M dan Ig G
Serologi
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Mencari bagian tertentu dari virus Dengue yang dapat menimbulkan penyakit baik berupa peptida ataupun asam nukleat
Deteksi Antigen
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tidak menunjukkan perbedaan pada infeksi primer maupun sekunder, dan beberapa kasus masih (+) pada fase konvalesen
marker deteksi dini infeksi dengue primer-sekunder
protein non-struktural yang disekresikan di permukaan sel terinfeksi
ditemukan dalam sirkulasi darah selama fase akut infeksi virus dengue
Antigen NS1
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glikoprotein 46-50 kDa terdapat di permukaan sel (membrane-associated /mNS1 ) & secreted form (sNS1)bentuk homodimer, partially hydrophobic nature bukan bagian dari struktur virussel yang terinfeksi di ekspresikan pada permukaan 1 dari 7 protein NS yang dihasilkan selama replikasi virusprotein yang disekresikan melalui cellular secretory pathway ke permukaan sel
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antigen NS1 dengue
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Dengue IgM Antibody IgM titre is higher in primary compared to
secondary infectionPeak at about 2 weeks and undetectable level
by 60 – 90 days. Recommended if dengue IgM is still
negative after day 7 in secondary dengue
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Dengue IgM Antibody Primary Dengue
55% detectable between 4-7 days100% after day 7
Secondary 78% after day 728% of secondary dengue infections were
undiagnosed when IgM was the only test performed
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Dengue IgG Antibody Primary and secondary dengue
IgG was detected in 100% after day 7 of illness
If Ig G was detected before day 7, that is a secondary infection
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Ig G Primary
WHO, 2009
Ig G Secondary
Ig M Secondary
Ig M Primary
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Interpretasi Hasil Ig m dan Ig GNo.Urut
Hasil InterpretasiIgG IgM
1. positif positif Dengue sekunder2. negatif positif Dengue primer3. positif negatif Dugaan dengue
sekunder
4. negatif Negatif Non-Dengue/ Primer amat awal. Retest setelah 4-7 hari
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Dengue Rapid testFalse positiveCross react with
Flavivirus-Non-flavivirus – malaria, leptospirosis,
toxoplasmosis, syphilisConnective Tissue Disorder
False negative low titre antigen/antibody
* ELISA METHODFOR DETERMINE
TITRE OF ANTIGEN/ANTIBOD
Y
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Keuntungan pemeriksaan antigen NS1deteksi infeksi dengue pada fase awaldemam (hari ke-1 sampai ke-5 sakit)tanpa menunggu terbentuknya antibodiSensitivitas pemeriksaan NS1 (88,7%) lebih tinggi
dibandingkan kultur virus (68,1%) dan PCR (66,7%).
Pemeriksaan IgG dan IgM antidenguetetap diperlukan untuk membedakan infeksi primer
atau infeksi sekunder
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KESIMPULAN
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ReferensiHandbook of Clinical Managemen of Dengue . WHO. 2012Nilai Novel Kombinasi Nilai Antingen NS1, Antibodi IgM dan IgG
Anti Dengue Dalam Memprediksi Outcome Infeksi Virus Dengue di Makassar. Rachmat Latief. 2010
Diagnosis cepat demam berdarah dengue. Danny wiradharma. 1999
Pemeriksaan Laboratorium Pada Demam Berdarah Dengue. Aryati. 2009
Laboratory Investigation. Salmah IdrisSerology Diagnosis of Dengue Infections. ECDC training
Workshop on laboratory diagnosis of dengue virus infectionsBerlin, 23‐27 January 2012
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TERIMA KASIH