DASAR VIROLOGIDASAR VIROLOGIdr. Ardhiyanti Puspita ratnadr. Ardhiyanti Puspita ratna

Laboratorium Biokimia Fakultas KedokteranLaboratorium Biokimia Fakultas KedokteranUniversitas Islam Al Azhar MataramUniversitas Islam Al Azhar Mataram

20132013

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Peristiwa bersejarah dalam bidang virologiPeristiwa bersejarah dalam bidang virologi

Th 1789; Penemuan vaksin cacar E. JennerTh 1789; Penemuan vaksin cacar E. Jenner Th 1892; Penemuan virus mozaik daun tembakauTh 1892; Penemuan virus mozaik daun tembakau Th 1918; Pandemi influenzaTh 1918; Pandemi influenza Th 1940; Penemuan Mikroskop ElektronTh 1940; Penemuan Mikroskop Elektron Th 1949; Penemuan teknik Tissue CultureTh 1949; Penemuan teknik Tissue Culture Th 1952; Penemuan DNA : Wartson & CrickTh 1952; Penemuan DNA : Wartson & Crick Th 1959; Penemuan vaksin polio (Sabin)Th 1959; Penemuan vaksin polio (Sabin) Th 1985; Kontroversi Penemuan virus HIVTh 1985; Kontroversi Penemuan virus HIV Th 1990; Teknik rekombinan vaksin (Hep B)Th 1990; Teknik rekombinan vaksin (Hep B) Th 1992; Aplikasi bio molekuler virus: HCVTh 1992; Aplikasi bio molekuler virus: HCV Th 2002; Multi Country Epidemi: SARSTh 2002; Multi Country Epidemi: SARS

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Sifat umum Virus:Sifat umum Virus:

Jasad renik hidup; ultra mikroskopikJasad renik hidup; ultra mikroskopik Parasit obligat intra selParasit obligat intra sel Partikel gen berdiri sendiriPartikel gen berdiri sendiri Dapat lolos saringan kuman (0,45 u)Dapat lolos saringan kuman (0,45 u) Dapat mensintesa ensimDapat mensintesa ensim Tropisme spesifik thdp hospesTropisme spesifik thdp hospes Bersifat antigenikBersifat antigenik Dapat dikristalkanDapat dikristalkan Bersifat onkogenikBersifat onkogenik Bersifat HemaglutinasiBersifat Hemaglutinasi Bersifat interferensiBersifat interferensi Dapat bermutasiDapat bermutasi 3

Bersifat toksikBersifat toksik Resisten thd antibiotikResisten thd antibiotik Dpt dihambat dg khemoterafiDpt dihambat dg khemoterafi Dpt direkayasa menjadi virus beda Dpt direkayasa menjadi virus beda

sifatsifat Dpt direkonstruksi menjadi vrius Dpt direkonstruksi menjadi vrius

lengkaplengkap Dpt membentuk plaques dan Dpt membentuk plaques dan

pockspocks 4

Teknik embrio Teknik embrio ayamayam Selaput khorio

alantoik

Embrio ayam umur 9-11 hr

rongga udara

POCKS

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Vero E6 cells inoculated with Oropharyngeal Specimens from Patients with SARS.The typical early cytopathic effect seen with coronavirus isolates from patients with SARS is shown in Panel A (x40). Infected Vero cells are shown reacting with the serum of a convalescent patient in an indirect fluorescence antibody assay in Panel B (x400).

Ksiazek et al. A novel Coronavirus associated with SARS. NEJM April 10, 2003

SARSV characterizationSARSV characterization

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Teknik biakan selTeknik biakan selPLAQUES virus pd biakan sel dlm media agar

PLAQUES

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Morfologi virusMorfologi virus Simetri kubus: ikosahedralSimetri kubus: ikosahedral Simetri heliks: spiralSimetri heliks: spiral Simetri kompleks: gab. kubus + heliksSimetri kompleks: gab. kubus + heliksA. Simetri kubus:A. Simetri kubus:

ikosahedron heksamer pentamerikosahedron heksamer pentamer berenvelopberenvelop

Contoh: Picornaviridae; polio, coxsackie, Contoh: Picornaviridae; polio, coxsackie, ECHOECHO 10

NP

M

Spike

Lipid bilayer

RNA

B. Simetri heliks

penampang lintang

penampang bujur

Contoh: Orthomyxoviridae; Paramyxoviridae,11

Simetri kompleks: Simetri kompleks:

bakteriofag bakteriofag

Simetri heliks

Simetri kubus

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13

14

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Figure 18.2 Viral structureFigure 18.2 Viral structure

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Figure 18.02x1 AdenovirusFigure 18.02x1 Adenovirus

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18

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Struktur virus:Struktur virus:

Unit kimiawi: asam nukleat Unit kimiawi: asam nukleat (DNA/RNA)(DNA/RNA)

Unit struktural: kapsomer Unit struktural: kapsomer kapsid kapsid nukleokapsid; nukleokapsid;

envelop (peplos): selubung protein envelop (peplos): selubung protein virusvirus

Unit morfologi: virion Unit morfologi: virion bentuk bentuk virus infektifvirus infektif

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Unit kimiawi:Unit kimiawi: Genom virus: suatu as. nukleat virus Genom virus: suatu as. nukleat virus

yang diwariskan: suatu rangkaian kode yang diwariskan: suatu rangkaian kode genetik yang mengekspresi protein genetik yang mengekspresi protein virus dengan bantuan komponen sel virus dengan bantuan komponen sel inang (host).inang (host).

Suatu ORF: open reading frameSuatu ORF: open reading frame

intron intron

5’ 3’

NCD struktural Non struktural (NS) NCD

hekson

Sistem Polisistronik: menghasilkan suatu rangkaian protein panjang 21

L

1 2 2-1 3 4 5 6 7

Cell penetration(+) genomic RNA (-) strand templateRNA# Protein2 ------> NS2

2-1 ------> HE

3 ------> S

4 ------> NS4

5 ------> NS5a, b

6 ------> M

7 ------> N

1 ------> Polgenomic RNA for new virions

Sistem Monosistronik: menghasilkan satu jenis protein

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Unit strukturalUnit struktural

Protein pada envelop virusProtein pada envelop virus Glikoprotein: hemaglutinin (bentuk Glikoprotein: hemaglutinin (bentuk

duri (spike); 10 nm). duri (spike); 10 nm). Lipoprotein: neuraminidase (ensim Lipoprotein: neuraminidase (ensim

utk replikasi virus)utk replikasi virus) Membran lipid bilayerMembran lipid bilayer

Unit morfologi: Unit morfologi: bentuk virionbentuk virion23

Replikasi virusReplikasi virusReplikasi virus DNA:Replikasi virus DNA:1.1. AdsorbsiAdsorbsi2.2. PenetrasiPenetrasi3.3. UncoatingUncoating4.4. Transkripsi awalTranskripsi awal5.5. Translasi awalTranslasi awal6.6. Transkripsi akhirTranskripsi akhir7.7. Translasi akhirTranslasi akhir8.8. Perakitan (morfogenesis)Perakitan (morfogenesis)9.9. Sel lisisSel lisis10.10. Virus baru (anak)Virus baru (anak)

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Replikasi virus DNAReplikasi virus DNA

Inti sel

virus

1

2

3

4

56

78

9

10 Contoh:

Adenovirus; virus Hepatitis B

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Replikasi virus RNA (RNA Replikasi virus RNA (RNA positif)positif)

RNA positif:

contoh: Picornavirus(virus polio)

Inti sel

translasi

genom +genom -

Protein virus

diferensiasi

polimerase; protein struktural

morfogenesis

progeni genom

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Replikasi virus RNA negatif:Replikasi virus RNA negatif:

pd. Orthomyxoviridae (virus pd. Orthomyxoviridae (virus influenza)influenza)

Replikasi virus RNA dg ensim Replikasi virus RNA dg ensim reverse transkriptase (HIV; virus reverse transkriptase (HIV; virus Hepatitis C)Hepatitis C)

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Replikasi retrovirusReplikasi retrovirus

RNAds DNA

Integrasi ke dlm kromosom inti sel host

transkripsi

translasi protein virus

morfogenesis29

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Reaksi thd. pengaruh kimia Reaksi thd. pengaruh kimia dan fisikadan fisika

Pelarut lemak: kloroform; ether Pelarut lemak: kloroform; ether ether sensitif dan ether resistenether sensitif dan ether resisten Deterjen: SDS: Sodium Dodesil SulfatDeterjen: SDS: Sodium Dodesil Sulfat

Triton X-100Triton X-100 Zat warna vital: penetrasi zw vital Zat warna vital: penetrasi zw vital

inaktif.inaktif.

ZW vital inaktif: HSV; vacciniaZW vital inaktif: HSV; vaccinia

inaktif sedang: Adenovirus; Reovirusinaktif sedang: Adenovirus; Reovirus

ZW vital resisten: poliovirusZW vital resisten: poliovirus Antibiotik: semua virus resisten antibiotikAntibiotik: semua virus resisten antibiotik

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KhemoteraKhemoterapipi: menghambat replikasi virus : menghambat replikasi virus dg cara menghambat kerja endg cara menghambat kerja enzzim virus.im virus.

Beberapa virus non responsif obat: VHB; Beberapa virus non responsif obat: VHB; VHC non responsif interferon; HIV non VHC non responsif interferon; HIV non responsif AZT.responsif AZT.

Desinfektan: semua virus sensitif thd khlor Desinfektan: semua virus sensitif thd khlor aktif (khlorin); formalin; HCl.aktif (khlorin); formalin; HCl.

Stabil dlm larutan garam anorganik:Stabil dlm larutan garam anorganik:

MgCl2: polio; enterovirus; reovirusMgCl2: polio; enterovirus; reovirus

MgSO4: influenza; campakMgSO4: influenza; campak

Na2SO4: herpes virusNa2SO4: herpes virus32

Pemanasan: 50 – 60 C; 30 menit Pemanasan: 50 – 60 C; 30 menit inaktif inaktif Penyimpanan: Penyimpanan: -20 C : enterovirus-20 C : enterovirus -70C s/d -120C (N2 cair/ CO2 kering utk -70C s/d -120C (N2 cair/ CO2 kering utk

transportasi); preservatif: gelatin; transportasi); preservatif: gelatin; glyserin.glyserin.

Liofilais: beku kering; kristal Liofilais: beku kering; kristal 4C. 4C. pH; virus tahan pd pH asam: enterovirus.pH; virus tahan pd pH asam: enterovirus. Radiasi sinar glb pendek: sinar UV; sinar Radiasi sinar glb pendek: sinar UV; sinar

XX as. nukleat rusak as. nukleat rusak mutasi/inaktif. mutasi/inaktif.

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KlasifikasiKlasifikasiDasar klasifikasi: Dasar klasifikasi: 1.1. Tipe as nukleat: DNA/RNATipe as nukleat: DNA/RNA2.2. Sifat as nukleat: ss/dsSifat as nukleat: ss/ds3.3. BM as nukleat: kD/kbBM as nukleat: kD/kb4.4. Simetri kapsid: ikosahedral; heliks; kompleksSimetri kapsid: ikosahedral; heliks; kompleks5.5. Tempat perakitan kapsid; inti sel/sitoplasmaTempat perakitan kapsid; inti sel/sitoplasma6.6. Selubung/envelop:/peplos: ada/tdk adaSelubung/envelop:/peplos: ada/tdk ada7.7. Tempat perakitan/morfogenesis: mbr sel. Tempat perakitan/morfogenesis: mbr sel.

sitoplasma; inti selsitoplasma; inti sel8.8. Diameter: virion; nukleokapsidDiameter: virion; nukleokapsid9.9. Kapsomer: jumlah kapsomer (pentamer; Kapsomer: jumlah kapsomer (pentamer;

heksamer)heksamer) 34

Klasifikasi yg sederhanaKlasifikasi yg sederhana

Virus DNA:Virus DNA:

As nukleat

simetri Diameter

(nm)

Famili

ss

ds

ds

ds

ds

ds

Ikosahedral

“

“

“

Komplex

“

18-26

45-55

70-90

100

230-400

22-42

Parvoviridae

Papovaviridae

Adenoviridae

Herpesviridae

Poxviridae

Hepadnaviridae

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Klasifikasi virus RNAKlasifikasi virus RNA

As nukleat

simetri Diameter

(nm)

Famili

ss

ds

ss

ss

ss

ss

ss

ss

ss

ss

Ikosahedral

“

“

Komplex

“

“

Heliks

“

“

“

20-30

60-80

30-90

50-300

80-130

s/d 100

90-160

80-120

150-300

70-175

Picornaviridae

Reoviridae

Togaviridae

Arenaviridae

Coronaviridae

Retroviridae

Bunyaviridae

Orthomyxoviridae

Paramyxoviridae

Rhabdoviridae36

Nomenklatur virusNomenklatur virus

Dasar:Dasar:I. Gambaran klinik: I. Gambaran klinik: measles; morbilli; small poxmeasles; morbilli; small pox Gambaran patologik; polimyelitis, hepatitis,dllGambaran patologik; polimyelitis, hepatitis,dll Gambaran epidemiologik: parotitis epidemikaGambaran epidemiologik: parotitis epidemikaII. Geografis: II. Geografis: NCDV (New Castle Diarrhea Virus);NCDV (New Castle Diarrhea Virus);

Murray valley Ensefalitis virus (MEV); West Nile Murray valley Ensefalitis virus (MEV); West Nile fever virus; Coxsackie virus, dll.fever virus; Coxsackie virus, dll.

III. Sifat utama partikel virus: III. Sifat utama partikel virus: OrthomyxovirusOrthomyxovirus mempunyai ensim neuraminidase (N-asetil mempunyai ensim neuraminidase (N-asetil neuraminat)neuraminat)

IV. Nama orang: IV. Nama orang: virus fibroma Shopevirus fibroma Shope, , strain virus: Edmonston; Schwarz; Sabin; Salk, strain virus: Edmonston; Schwarz; Sabin; Salk,

Saukett, dll.Saukett, dll.37

V. Huruf awal: REO (Respiratory Enteric V. Huruf awal: REO (Respiratory Enteric OrphanOrphan

ARD: Acute Respiratory DisaesesARD: Acute Respiratory Disaeses

VI. Gabungan kata:VI. Gabungan kata:

PicoRNAvirus: virus RNA kecil (25 nm)PicoRNAvirus: virus RNA kecil (25 nm)

HepaDNAvirus: virus DNA dlm heparHepaDNAvirus: virus DNA dlm hepar

Papovavirus: papiloma-polioma-vakuola-Papovavirus: papiloma-polioma-vakuola-

virusvirus

ARBOvirus: Arthrophode Born VirusARBOvirus: Arthrophode Born Virus

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Lysogenic versus Lytic Cycles of Lysogenic versus Lytic Cycles of BacteriophageBacteriophage

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Life Cycle Papovirus ( Virus Life Cycle Papovirus ( Virus DNA)DNA)

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Retroviruses Convert RNA into DNA via Reverse TranscriptaseRetroviruses Convert RNA into DNA via Reverse Transcriptase

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Structure of Influenza Structure of Influenza VirusVirus

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Viruses and CancerViruses and Cancer

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Web LinksAll the Virology on the WWW

(http://www.tulane.edu/~dmsander/garryfavweb.html) All the Virology on the WWW "seeks to be the best single site for Virology information on the Internet. We have collected all the virology related Web sites that might be of interest to our fellow virologists, and others interested in learning more about viruses".

The Big Picture Book of Viruses(http://www.tulane.edu/~dmsander/Big_Virology/BVFamilyGenome.html)

The Big Picture Book of Viruses is "intended to serve as both a catalog of virus pictures on the WWW and as an educational resource to those seeking more information about viruses".

Electron microsopic images of Human Viruses(http://www.uct.ac.za/depts/mmi/stannard/linda.html) Electron microsopic images of Human Viruses - Linda Stannard's "illustrated tutorial on the morphology of most of the clinically significant viruses. The section on Hepatitis B virus is especially recommended".

Visualizations of Viruses at the University of Wisconsin - Madison(http://www.bocklabs.wisc.edu/virusviztop.html)

The Index Virum(http://life.anu.edu.au/viruses/Ictv/index.html) The Index Virum presents lists of virus taxa that reflect the currently approved classification of the International Committee on Taxonomy of Viruses (ICTV).

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Terima KasihTerima Kasih

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