By: Dr.Malak El-Hazmi Dr. Abdulkarim Alhetheel
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Transcript of By: Dr.Malak El-Hazmi Dr. Abdulkarim Alhetheel
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By: Dr.Malak El-Hazmi
Dr. Abdulkarim Alhetheel
( Foundation Block , Microbiology : 2011 )
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OBJECTIVES
General characteristics of viruses.Structure & symmetry of viruses.Classification of viruses.Steps of virus replication . laboratory diagnosis of viral infections.
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Properties of Microorganismscharacteristic Parasites Fungi Bacteria Viruses
Cell Yes Yes Yes No
Type of nucleus
Eukaryotic Eukaryotic Prokaryotic -----
Nucleic acid Both DNA& RNA
Both DNA& RNA
Both DNA& RNA
DNA or RNA
Ribosomes Present Present Present Absent
Mitochondria Present Present Absent Absent
Replication Mitosis Budding ormitosis
Binary fission
special
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Characteristics of viruses Acellular organisms Tiny particles
• Internal core• Protein coat• Some Vs have lipoprotein mb
Obligate intracellular organisms Replicate in a manner diff from cells ( 1V many Vs )
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Size ; 20-300 nm
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Viral Structure 1-Viral genome
2-Capsid
3-Envelope
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Viral Structure1-Viral genome
RNA (Ribonucleic acid) All RNA Vs have ss except Reoviruses single / multiple (+) polarity (-) polarity
DNA (Deoxyribonucleic acid) All DNA Vs have ds except Parvoviruses Single molecule
or
All Vs are haploid ,except retroviruses are diploid
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Viral structure2-Capsid a protein coat Subunits (capsomeres) Genome (NA) + capsid = nucleocapsid Function;
Protects NA Facilitates its entry into cell
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Symmetrybased on arrangement of capsomeres
Cubic symmetry ( Icosahederal )
Helical symmetry Complex symmetry
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Symmetrybased on arrangement of capsomeres
1-Cubic symmetry ( Icosahedral )
Adenovirus Herpesvirus
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Symmetrybased on arrangement of capsomeres
2- Helical symmetry
3- Complex symmetry poxviruses
Elongated (filoviruses)
Pleomorphic ( influenza v.)
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Viral structure3-Envelope Lipoprotein mb(host lipid ,virus specific protein ) Budding Envelope is derived from cell mb except herpesviruses from nuclear mb Enveloped Vs are more sensitive to heat ,dry & ether than nonenveloped Vs Glycoprotein attaches to host cell receptor
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Viral proteinsThe outer viral ps Mediate attachment to specific Rs Induce neutralizing Abs Target of Abs The internal viral ps Structural ps ( capsid ps of enveloped Vs ) Nonstructural ps ( enzymes)
All ssRNA Vs (-) polarity have transcriptase ( RNA dependent RNA polymerase) inside virions RetroVs & HBV contain reverse transcriptase
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Classification of viruses Type of NA The no. of strand The polarity of viral genome The presence or absence of envelope Type of symmetry
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Medically Important Viruses
ComplexPoxviridae
Icosahedral HerpesviridaeHepadnaviridae
IcosahedralAdenoviridaePapovaviridae
Enveloped
RNADNA
Nonenveloped
Single-stranded double-stranded
IcosahedralParvoviridae
Nonenveloped
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HelicalOrthomyxoviridaeParamyxoviridaeRhabdoviridaeFiloviridaeBunyaviridaeArenaviridae
HelicalCoronaviridaeIcosahedral Togaviridae Flaviviridae Retroviridae
IcosahedralReoviridae
Enveloped
RNADNA
Nonenveloped
Medically Important Viruses
double-strandedSingle-stranded
Nonenveloped
IcosahedralPicornaviridae Hepeviridae Caliciviridae Astroviridae
Pos- strandNeg - strand
Enveloped
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Replication Adsorption
(Attachment) Penetration Uncoating Synthesis of viral
components• mRNA• Viral proteins• NA
Assembly Release Viral growth cycle
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Attachment site ; - glycoprotein
- folding in the capsid proteins.
Adsorption
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Penetration2-Endocytosis
Viral envelope fuses with endosome mb Nonenveloped V.
lysis ,pore
1-Fusion
(enveloped Vs )
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Replication Adsorption (Attachment) Penetration Uncoating Release of viral genome - cytoplasm - nucleus
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Synthesis of viral components mRNA
Viral genome transcription mRNA +ssRNA acts directly
Viral proteins mRNA translation viral proteins cell ribosome - enzymes - structural ps
replication of viral genome
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Replication Adsorption (Attachement) Penetration Uncoating Synthesis of viral components
• mRNA• Viral proteins• NA
Assembly NA + V. proteins = Virions Release
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Release 1-Budding(enveloped Vs)-cell mb*-nuclear mb (herpesVs)
2- Cell lysis or rupture (nonenveloped)
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laboratory diagnosis of viral infections
Microscopic examination.Cell culture.Serological tests .Detection of viral Ag.Molecular method .
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Microscopic examinationLight microscopy; Histological appearance Ex. Inclusion bodies Owl’s eye (CMV)Electron microscopy;
Morphology& size of virions Ex. Dx of viral GE ,rotav. , adenov. Dx of skin lesion caused by herpesv, poxv. It is replaced by Ag detection & molecular tests
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Electron micrographs
Poxvirus
Rotavirus
Herpesvirus
Adenovirus
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Virus cultivationLaboratory animalEmbryonated eggCell culture
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Cell culture No of sub passages
Primary C/C 1 or 2
Diploid C/C[semi continuous]
20 to 50
Continuous cell line
Indefinite
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Variation in Sensitivity of cell cultures to infection by viruses commonly isolated in clinical virology laboratories
VirusCell culturea
PMK HDF HEp-2
RNA virus
DNA virus
Enterovirus Rhinovirus Influenza virus RSV
Adenovirus HSV VZV CMV
+++ ++ +/-+ +++ ++++ + -++ + +++
+ ++ ++++ ++ +++ +++ -- +++ -
PMK, primary MK. Degree of sensitivity: +++, highly sensitive;++, moderately sensitive; +, low sensitivity; +/-, variable; -, not sensitive
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Detection of viral growthCytopathic effects
IFOther
Uninfected cc Cell rounding Syncytium
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Problems with cell culture ;
Long incubation Sensitivity is variable Susceptible to bacterial contamination Some Vs do not grow in c/c ex. HCV
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Rapid culture technique Shell Vial AssayDetect viral antigens1-3 days
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Serological test;Antigen detection;
sample virus test Nasopharyngeal aspirate Influenza V. IF Skin scrapings HSV IF Faeces Rotavirus ELISA Blood HBV(HBsAg) ELISA
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Serological test;
Ex of techniques Complement fixation test (CFT) Immunofluorescence (IF) Enzyme- linked immunosorbent assay (ELISA)
Antibody detection;
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Immunofluorescence ; IF
A- Direct Ag detection;
Sample (Ag)
B- Indirect Ab detection;
Sample (Ab)
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ELISAAb detection Ag detection
Indirect ELISA for Ab detection ; coloured wells indicate reactivity
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Molecular test; Polymerase chain reaction (PCR)
Amplification tech. Viral genome
Uses; Dx Monitoring response to Rx
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Reference book and the relevant page numbers
Review of Medical Microbiology and Immunology
By: Warren Levinson . 10th Edition, 2008.
Pages;192-195,199-207, 216-220,233-235.
Lippincott’s Illustrated Reviews: Microbiology By: Richard A.Harvey ,
Pamela C Champe & Bruce D. Fisher
2nd Edition, 2007 . Pages;233-242
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