Introduction: - Viruses are obligate intracellular parasites replicating only in living cells while inert extracellularly. There sizes range from 20-30nm. The complete infectious virus particle is caje, virion which is made up of viral genome (either RNA or DNA) enclosed in a protein coat calleg capsid. In turn RNA viruses can be +ve stranded and —ve stranded. Viruses lack specific enayre; required for replication and uses the host synthetic machinery for this purpose to produce specific macro molecules required to form viral progeny. During this, the host cell may have little effect or it may result in cell damage and death. Comparison between Viruses and Cells N= Type of nucleic acid DNA or RNA but not both | DNA and RNA Protein Few Many “ Lipoprotein membrane Envelop present in some Cell membrane present inl viruses cells - Ribosomes _ Absent Present Mitochondria | Absent Present in Eukaryotic cells oo only Enzymes __None or few Many Multiplication by binary "No ves fission or mitosis | | Scanned with CamScaIpere are 4 components in describing of viral eee Nuc ae Jgructure (Figure 16-1): - fon) coy Jenene 1) Viral Genome (Either DNA or RNA) 2) Capsid 3) ) Viral Genome: - Asc ctin A iral Envelop 4) Viral Nucleocapsid it can be single stranded or double stranded, Nucteocapsid linear or capsular and segmented or lunsegmented. ‘eit is associated with viral specific enzymes or e lother proteins within the virion or both. Capsid: - : is composed of structural units called “Capsomers” which are aggregates of viral specific lypeptides, its structure determine the viral “symmetr eg, Orthomyxoviras)ortCornplex (e.g. Powvrus)) Cubic symmetry seen osahedral type. ~~ Upsd envelope Grcoprotein spikes FIGURE 16-1 “Viral Structure” ‘A: Nonenveloped virus with an icosahedral nucleocapsid, : Enveloped virus with an helical nu [Function of capsid include protection of viral genome and to act as a site of receptors of ‘faked virus to intimate infection. lt act as a stimulus for antibody production and is the site of antigenic determinacy which is important for serologic tests. 3) Viral Envelop: - sembly of the virus. It is usually host cell plasma membrane and sometimes nuclear Dembrane or endoplasmic reticulum too, Viral glycoproteins are often embedded in membrane, used for binding to host cells and also antigens for immune system. Allnaked Viruses have icosahedral capsid whereas envelopd have icosahedral or helical. 4) Viral Nucleocapsid: - This refers to the protein nucleic acid complex (capsid and enclosed viral genome), ‘eptesenting th . e packaged from the virus. / “Danson et oeaaata uses e.g. envelopd viruses “*Thisterm is commonly used in case of structurally complex vir Scanned With CamScaInstant Microbiolog) a ocapsid). env ot part of nucle _. eee ‘is identical to virion. «a (Lipid , es, nucleocapsi in case of naked virus' Prions: - roteins. They have no DNA Prions are infectious particles composed entirely of PI et ey have no DNA or RNa, They cause disease such a creutzfeldt-jakob disease/and kuru in humansand madcow di and scrapie in animals) These diseases are called transmissible spongiform encephalopathg, re pearance of the brain seen in these diseases, The term spongiform refers to sponge like ap holes of the sponge are vacuoles resulting from dead neurons. Prion are encoded by a cellular gene. When these proteins are normal_alpha-helix configuration, they are nonpathogenic, but when their configuration changes to a beta plz isruy eur and causes get, th regate into filaments, which Prions are highly resistant to inactivation by UV light, heat and other inactivating agents. Ass result, they have been inadvertently transmitted by human growth hormone and neurosurga} instruments. As they are normal human proteins, they do not elicit an inflammatory response or an antte response in humans. Comparison of prions and conventional viruses eee Particle containing nucleic acid __| No Yes Particle containing proteins Yes, encode by cellular Yes, encoded by viral proteins | Inactivated rapidly by UV light or | No Yes _heat Appearance on electron Filamentous rods (amyloid | icosahedral or heli microscope like) symmetry Infection induce antibody No Yes Infection induce inflammation No Yes SHAPES OF VIRUSES: - The shapes of viruses are frequently referred to in colloquial a term e.g. spheres, £045. or bricks but in reality they are complex structures of precise geometric symmetry: TM Of virus particle is determined by the arrangement of the repeating subunits that fom Coat (capsid) of the virus (Figure 16-2). ‘ Scanned with CamScaInstant Microbiology Herpesvirus Poxvirus mw €¢ e : T ‘Adenovirus Papovavirus Hepadnavirus Parvovirus DNA viruses oo Coo ° Paramyxovirus Orthomyxovirus Coronavirus Arenavirus Retrovirus Filovirus Picornavirus Rhabdovirus Togavirus Bunyavirus RNA viruses FIGURE 16-2 Shapes of Viruses Scanned with CamScza in living cells and involves the host cell enzymes. itmay be complete n sme cels Abort infection] yang detective walt During viral replication, normal host DNA, RNA or proteins can not be synthesized, leading to damage to host cell which can manifest as cell death. The steps of viral replication are: - 1) Attachment: »—~ - It involves interaction between viral receptors and host cell receptors. This phase determines viral tropism. 2) Penetrations This can occur by endocytosis (yiropexix), direct penetration or fusion of virus envelop with lasma rane of host cell. / 3) Uncoating: - Release of the genome from the capsid into cytoplasm or nucleus. 4) Viral gene expression: - ruses require virus specific messenger RNA to syntiouice virus specific proteins, the viral mRNA. nome and also have an RNA polymers: (Rotavirus has a double stranded RNA genome RNA Viruses ] Some RNA viruses, such asoliovirusyhave a Positive polarity RNA genome that serves as the mRNA. Other viruses such asi negative polarity RNA genome and have an RNA polymerase in the virion that synthesizes DNA Viruses uses such as(herpes viruses) 22. and papilomavirusés have 2 | jou oded DN me and use the | ‘NA polymerase to synthesize the A uses havea doubl : the virion that synthesizes the viral m&N and has an RNA polymerase in the virion that | Poxvirus has an RNA polymerase in the ¥ synthesizes the viral mRNA. because they replicate in the cytoplas Retrovirus such as HiV;have a Positive not have access to the host cell RNA polarity RNA genome and have a DNA Polymerase in the virion that synthesizes a DNA copy of RNA genome. This DNA is the template used by the host cell RNA, Polymerase to synthesize the viral mRNA polymerase in the nucleus. ————— Scanned with cithecter 17: Viral Replication Instant Microbiology 5) Assembly and release: - elopd viruses acqui allenvelopd ire their envelop by budding through the e membrane through the external cell membrane a: e cell ex i whi ri Hea ialan they exit the cell except herpes viruses which acquire their envelop by budd ; udding through nuclear s. membrane. The matrix protein mediate the interaction of the nucle d 0c i leer: jpsid within the envelop Itis the process by which viral DNA becomes i Se oda Late TUNA integrated into host cell DNA, replication stops 5 Sn : 2 I ONA is, damaged by, for example UV light, viral DNA is excised from 7 and progeny vituses are made, The integrated viral DNA is called Ba prophal re. Eat cell carrying a prophage can acquire new traits such as the ability to produce exotoxin such as diphtheria toxin. Transduction is the process by which virus carry nes from one cell to another. Lysogenic conversion is the term used to indicate that the cell has acquired a new trait as a result of the integrated prophage. VIRUS GROWTH CURVE: - irus infects a cell and hundreds of progeny ind explains the rapid spread of virus from cell to cell, .d within the infected cell. It y virions are produced within hours. This is 2 One femarkable amplification a! The eclipse period is the time when no virus particles are detecte' occurs soon after the cell is infected (Figure 17-1). both morphologic and functional, Cytopathic effect is the term used to describe damage, the presence of virus in the patient's ~ Inficted on the ce[L bythe virus, In the clinical laboratory, specimen is often detected by seeing 2 CPE in cell culture. | Viral nucleic acid (=~ -) e789 wt | oot Soa 4 8 | Eclipse period +— Rise period | L Hours FIGURE 17-1 Viral Growth Curve | - Scanned with CamScagenic variants and drug resistant variants. Mutat Mutation in viral genome can produce anti (weakened) variants that can not cause disease but retain ther can also produce attenuated licate at low temperature but not. at high temperatu in one.of the vaccines against ths, Temperature sensitive mutants can re| uenza virus are used i Temperature sensitive mutants of infl disease. ween viruses occurs through 3 different notypic mixing Interaction bet 1) Reassortment 2) Complementation 3) Phe ent of genome RNA of influenza virus is important inthe Reassortment (exchange) of seam athogenesis of the worldwide epidemics caused by this virus- duce a protein that can be used by other vin pathogenesis of Ie Complementation occurs when one vtus prodek AFT «rally important example & Hepatitis whch us heise ‘of hepattst virus as its outer coat protein. wes 5 er ten vii eS aerate ocboth pret nse. Thcan endo fe OHSAS with the & infect cells of species that ordinarily parental virus could nos. ON OF MEDICALLY IMPORTANT VIRUSES 0 components uses is based on chemical and morphologic criteria. The tw’ phenomenon: - CLASSIFICAT! f vir" ‘n classification are: - lecular weight and structure). etry and whether it is envelopd). Classification © the virus used i 1) Nucleic acid (its mo! 2) Capsid (Its size and symm: DNA Viruses: ~ etry: AILDNA viruses contain hedral symm! All have icos@! AlLDNA viruses Fe) icate in the nucleus. etry except for poxvirus which has complex symm —_ Scanned with CamSymmetry Icosahedral Icosahedral_ | DNA 'Structire DS, circular, | supercoiled Instant Mic, Tobiology LUCIE Important wi Icosahedral DS, circular, Human papillomavirus 7 supercoiled No | !cosahedral | DS, linear {RdenoviruS Yes Icosahedral | DS, incomplete | Hepatitis B virus | circular Yes Icosahedral | DS, linear CHECDES SIMpIEXviTs, tomes EBV Yes Complex DS, linear (Contagiosum virus) Envelop | Capsid sce Symmetry Icosahedral (Rerpesvia} andfeovind) ept forfetrovirus and orthomyxovi RNA Structure: SS, linear, - nonsegmented, positive polarity Medically Ui) eget No Icosahedral 5S, linear, nonsegmented, positive polarity Hep E virus) No Icosahedral SS, linear, nonsegmented, itive polarity No Icosahedral near, 10 or 11 segments _ Yes Icosahedral_ 55, linear, nonsegmented, Scanned with CamSca: Viral Genetic and Clasug. positive polarity Icosahedral | SS, linear, Rubella vin ~~ nonsegmented, | | | _ positive polarity | Retrovirus Yes Icosahedral_| SS, linear, 2identical (HIV, Ruman Tce | strand (diploid) , leukemia vin positive polarity SS, linear, 8 segments, ((pfluenza virus Togavirus “Yes | Orthomyxovirus | Yes Helical | negative polarity Paramyxovirus _| Yes Helical SS, linear, Méasles, mump>~ nonsegmented, _ negative polarity | Rhabdovirus Yes Helical 5, linear, Rabies virus> | nonsegmented, negative polarity } | Filovirus Yes Helical SS, linear, bola virus, nonsegmented, (Marbung viru) negative polarity Yes Helical SS, linear, -+Coronavirus Coronavirus ——— nonsegmented, — positive polarity Yes Helical SS, circular, 2 segments | lassa fever vin) Arenavirus | with cohesive end, negative polarity. Yes Helical $5, circular, 3 segments | Gantavirus font Bunyavirus ta = with cohesive end, €ncephalitis v! negative polarity. — Yes Uncertain —_| SS, circular, closed Hepatitis delta¥™ circle, negative polarity LL at Deltavirus Scanned with CamScaPye ieee te) The ability of viruses to causes disease can be viewed on two distinct levels: 1) The infected cell 2) The infected patient - The infected cell: - f infected cells is probably caused by inhibition of Translation of viral mRNA into viral proteins preempts the ribosomes preventing synthesis of cellular protein synthesis. A st 3 o. > = cellular proteins. ~ inclusion bodies are aggregates of virion in specific location in the cell that are useful for \jisratory diagnosis. Two important examples are,negri bodies in the cytoplasm of rabies virus cleus of cytomegalovirus infected cells. infected cells and owl's eye inclusion, in the nu m when cells are infected with Certain viruses notably ~ Multinucleated giant cells are for herpesvirus and paramyxovirus such as respiratory synctial virus. _ oytopathic effect is a visual or functional change in infected cells typically associa! enic viruses. | death.of cells. Malignant transformation occurs when cells are infected with oncog' d cells appear visually and functionally normal, yet are producing large number of progeny Wf. The infected patient: - | Viral infection in the person typica Hincubation period 2yProdromal period) 3) The main portals of entry-are res} irato! asioss placenta and via bloo o offsprint ‘transmission (e.g. fecal-oral, respiratory, | Most serious viral infections are system" i.e. viru: 'd to the portal of entry such as common cold, which | a Hn TT sesipemcencam involves only upper respirato! . Mechanism z |!amunopathogenesis is the process by which the symptoms of viral diseases are caused b [iteoun ‘tem rather than B the killing cell directly by the virus. One type of | immunopathogenesi: i my wnesis is the killin of virus infected cell by the attack of cytotoxic T-cells that | a E - Scanned with CamScz ted with the lly has four stages: - Speicific illness period Recovery perio’. cointestinal and geni ‘al tract bi astr h sh 'd are important as well. ig is called vertical tran aerosol, insect bite) are horizontal transi 's travel from portal of entry via blood to smission while al! other moces of ‘Transmission from mother t ssion.~~ Instant Microbiology ical Rae recognize viral antigen on the cell surface, pamage to the liver caused by fe tis viruse i occurs by this mechanism. Another is the formation offvirus antibody complexesythat are i occurs by this mechanism. An denositedin tissues, Arthritis associated with parvovirus 419 pr cul infection occurs by this mechanism. virus can evade host defenses by producing multiple antigens, thereby avoiding activation by antibodies and by reducing the synthesis of class | MHC proteins, thereby decreasing the ability of cell to present viral antigens and blunting the ability of cytotoxic T cells to kill the virus infected cells. Virus can also produce receptors for immune mediators, such as IL-1 and TNF, thereby preventing the ability of these mediators to activate antiviral processes. Persistent viral infections: - In certain instances, the virus persist for long periods either intact or in the form of subviral 7 component (e.g. genome).The mechanism that may play role in persistent of viruses include; - 1) Integration of DNA provirus into host cell DNA as occurs in retroviruses, 2) Immune tolerance because neutralizing antibodies is not formed. 3) Formation of virus antibody complex which remain infectious. 4) Location within immunologically sheltered “sanctuary” (e.g. the brain) 5) Rapid antigenic variations 6) Spread from cell to cell without an extracellular phase so the virus is not exposed to antibody. a 7) Immunosuppression as in acquired AIDS. There are 3 types of persistent virus infection of clinical important: - 1) Carrier state: - It refers t le who produce virus for long period of time and can serves a sousce of infection for others. The carrier state that is frequently associated withfhepatitis 2) Latent infections: - These are those infections that are not producing virus at present time but can be reactivated at a subsequent time. The latent infections are frequently associated with Kerpes simplex viruare of medically important, 3) Slow virus infection: It refers to those diseases with a long incubation period, often- viruses whereas others, such as Creutzfeldt-Jakob disonse are caused by prions he bral." main site of these diseases, HOST DEFENSE: Host defenses against virus fall into 2 categories: - 1) Non-specific, of which most important is interferons and natural killer cells. 2) Specific which includes humeral and cell mediated immunity. Scanned with carhce) Non-specific defenses interferons: - s and double st Tanded RNA are the most potent inducer of interferon. Many viruses cuce interferon and many viruses are inhibited by interferon (i.e. neither the induction nor tion of interferon is specific). Interferon act by binding to the receptor on the cell surface als the cell to synthesize ribonuclease protein kinase and oligo A synthetase in an ctive form. Double stranded RNA made by the infecting virus activates these proteins. ron does not enter the cell and have no effect on extracellular viruses. Interferon inhibits lication by knocking protein synthesis, primarily by degrading mRNA and by_ ing elongated factor-2. Alpha and beta interferon have stronger antivigal action than 2 interferon. The later acts primarily as an interleukin that activate: tural killer cells: - re lymphocytes that destroy cells infected by many different viruses. Nk cells do not ic receptor is i |B lymphocytes. Rather, NK cells recognize 2 destroy cells that do not display class | MHC protein on the surface. They kill cells by the gos mechanisms as do cytotoxic T cells (i.e. by secreting perforins and granzymes). non specific defense are Phagocytosis, Z-Defensin, Apolipoprotien B RNA editing enzyme Circumcision and Factors that modify host defenses. 3G), Fever, Mucociliary clearance, Specific defenses i ive ienrmunity to viral infection is mediated by both antibodies and cytotoxic T cells. It can be “ed either by exposure to the virus or by immunization to the viral vaccine. ve immunity consists of antibodies preformed in another person or animal. ration of active immunity is longer than passive immunity. Active immunity is measured (2's, whereas passive immunity lasts a few weeks toa few months. ve immunity is effective immediately, whereas active immunity takes 7 to 10 days in “ay response and 3 to 5 days in secondary response to stimulate detectable amount of “immunity is the protection of an individual that results from i ity in many other Tiber of the population (the herd) that interrupts transmission of the virus to the individual. ‘7 munity can be achieved either by immunization or by natural infection of a sufficiently “centage of the population.tant Microbiology -sis and Host Defence Ins: 4) Non-spec' ic defenses interferon: yeuses and double stranded RNA are the most potent inducer of interferon. Many viruses | inauce interferon and many viruses are inhibited by interferon (i.e. neither the induction nor | iye action of interferon is specific), Interferon act by binding to the receptor on the cell surface le signals the cell to synthesize ribonuclease protein kinase and oligo A synthetase in an | active form. Double stranded RNA made by the infecting virus activates these proteins. interferon does not enter the cell and have no effect on extracellular viruses. Interferon inhibits, | inactivating elongated factor-2. Alpha and beta interferon have stronger antiviral action than jena interferon. The later acts primarily as an interleukin that activates. hage. | Natural killer cells: - Nicolls are lymphocytes that destroy cells infected by many different viruses. NK cells do not fave antigenic receptor on their surface, unlike T and B lymphocytes. Rather, NK cells recognize |and destroy cells that do not display class | MHC protein on the surface. They kill cells by the ms as do cytotoxic T cells (i.e. by secretiny erforins and granzymes). (ther non specific defense are Phagocytosis, Z-Defensin, Apolipoprotien B RNA editing enzyme P08EC3G), Fever, Mucociliary clearance, Circumcision and Factors that modify host defenses. | |) specific defenses ive immunity to viral infection is mediated by both an lected either by exposure to the virus or by immunization to the viral vaccine. Passive immunity consists of antibodies preformed in another person or animal. Be duration of active immunity is longer than passive immunity. Active immunity is measured i, Ye2rs, whereas passive immunity lasts a few weeks to a few months. | sssive immunity is effective immediately, whereas active immunity takes 7 to 10 days in Barrsesronse and 3 to 5 days in secondary response to stimulate detectable amount of Ptivody. tibodies and cytotoxic T cells. It can be Her non 7 rua the protection of an individual that results from immunity in many other hee Is of the population (the herd) that interrupts transmission of the virus to the individual. |? immunity can be achieved either by immunization or by natural infection of a sufficiently i Pgh _*"Rercentage of the population. Scanned with CamScazLABORATORY DIAGNOSIS USED FOR VIRUS Diagnostic virology involves one or all of the three basic techniques that are 1) Viral isolation 2) Direct demonstration of virus 3) Viral nucleic acid or viral antigen in specimen or in serologic testing. 1) Viral Isolation: - In this viruses are cultivated from clinical specimens into tissue culture cells, embryonatede or animal host. Viral replication is identified in live infected tissue culture by observing pathic effect (e.g. prokaryotic formation and adhesion of RBCs to infect clusion bodies. characteristic cyto} cells called hemeadsorption) and in fixed tissue culture by observing int 2) Direct Examination: - It can be done on sections of tissue biop: ies, tissue smear, blood, CSF, saliva, urine, feces or 4 throat swab. The following methods can be used in this technique. IMMUNOHISTOCHEMICAL NUCLEIC ACID SOLID PHASE Sis Petree IMMUNOASSAY Fixed or fresh specimens and Detection of viral DNA or _| Viral antigens are detect chemically (e.g. fluorscein) or RNA sequence in nucleic by using specific RIA, oF enzymatically (e.g. peroxidase) acid extracted from ELISA technique especi labeled antibodies are used to specimen is done by PCR, useful in rotavirus and identify viral specific antigen. DOT blot hybridization or —_| hepatitis A virus. nucleic acid probes especially used in CMV or HIV. 3) Serologic Testi It detect the titer of specific antibodies. It can be done by viral neutralization test (cert antiviral antibodies neutralize the cytopathic effect of virus), hemagglutination tests or solid phase immunoassay. UT EZ Neca S There are only few drugs that are useful against viral infections thereby prevention of ¥ fain iral infections can be achieved by the use of vaccines that induces active immunity. There are three types of vaccines that can be used: - 1)4Gve; attenuated vaccine?) killed virus a a{Recombinant vaccine’) Scanned with catpter 20: Laboratory Diagnosis & Viral Vaccines e, attenuated vaccines: - is inactive strain) rarely can produce clinical disease. It cannot be given to munocompromised patients. rotection (no boosters) ZV), Sabin's polio viru; (@MP)andGntranasal influenza led virus vaccines : has no risk of infection, require less immune response (boosters often required). |t is used for bidk injected influends, mbinant vaccines: - e vaccine protein gene is inserted into a virus or cells in culture. When carrier virus or cell 's so vaccine protein is created and immune system will recognize the expressed protein. It used for(Hepatitis B vaccine» Recombinant HbsAg and HPV: Recombinant proteins types 6, 11 6, and 18. ives long la Scanned with CamSca