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GENERAL VIROLOGYGENERAL VIROLOGY

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Viruses

The smallest infectious agents(20-300 nm

onl! seen un"e#

Elect#on $ic#osco%e

(e&ce%t 'o&i#uses

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Elect#on $ic#osco%e

- The source of illumination is a beam of electrons

-Electromagnetic lenses

- Magnifcationis )00*000 o# mo#e

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Elect#on $ic#osco%!

+acte#ial

,m#iae

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Structure of Viruses

Virus particle (virion)

Structure of viruses

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7

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8

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Vi#al .!mmet#!

)* Icosahe"#al s!mmet#!/e.g., all DNA viruses, exceptpoxviruses & some RNA viruses

2* elical s!mmet#!/

e.g., many RNA viruses (e.g. rabiesvirus

3* 1om%le& s!mmet#!/

e.g., poxviruses (bric!-s"ape#

Ico

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$cosa"e#ral %ymmetry

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Rotai#us 1alicii#usAst#oi#us

Icosahedral Symmetry

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elical

.!mmet#!

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Complex symmetry

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ie#ences et4een i#uses5 acte#ia

)* The! a#e oligato#! int#acellula#%a#asites

"ey can not be cultivate# on artifcialculture

me#ia. can only replicate insi#e living cells.

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2* Vi#uses contain onl! one t!%e ofnucleic aci"

(NA o# RNA6 nee# oth*

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3* The! can note cultiate" on

a#ti,cial cultu#e me"ia*

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7* The! a#e not susce%tile to

antiacte#ial agents*

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iagnosis of i#alinfections

A- i#ectmetho"s

+- In"i#ect metho"

I* i#ect"etection ofi#uses an" 8

o# thei#com%onents

$$. $solationo' viruses

I*.e#olog

!

$$. %!intests

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I* i#ect "etection of i#uses 5 8 o#thei# com%onents/

)* Light mic#osco%!

)xamination o' large virusesas *oxviruses

Detection o' giant cells

in +erpes in'ectionDetection o' inclusion bo#ies

e.g. Negri bo#ies in nerve

cells in rabies

A- i#ectmetho"s

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2* Elect#on mic#osco%! (E$arge number o' viruses in t"e

sample.%ie an# s"ape o' viruses.

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3* Immunoelect#on mic#osco%!(IE$/

%ample (un!non virus / !nonspecifc antibo#y

agg#egationo' un!non virusparticles

e.g. "epatitis A virus in stools

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7* 9luo#escent mic#osco%!/

Direct immuno0uorescent antibo#y tec"ni1ue ($2

e.g. #iagnosis o' rabies in brain smears.

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:* Immunoassa!s /

2or #etection o' t"e virus antigens by )$%A 3R$A e.g. "epatitis 4 antigens in bloo#

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ELI.A

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;* Nucleic aci" h!#i"i

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>* 'ol!me#ase chain #eaction ('1R

Amplifcationo' a specifc se1uence o' nucleic aci#

Detection e.g. by gel electrop"oresis.

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A- i#ectmetho"sII*Isolation of i#uses/

5bligatory intracellular parasites

Replicate only in livingsusceptible cells

a* Lao#ato#! animals

* Em#!onate" eggs

c* 1ell cultu#e (tissuecultu#e
http://images.google.com/imgres?imgurl=http://mrshenri.files.wordpress.com/2009/07/chimpanzee-head.jpg&imgrefurl=http://mrshenri.wordpress.com/2009/07/19/banana-test/&usg=__FBG4kgBvX0kam4NrM_SKfSmmnZk=&h=1001&w=1001&sz=271&hl=en&start=5&itbs=1&tbnid=tA5fc1q4e6LLZM:&tbnh=149&tbnw=149&prev=/images%3Fq%3Dchimpanzee%26hl%3Den%26tbs%3Disch:1

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?se" in/

$solation o' coxsac!ie viruses by inoculation in

4hite suc@ling mice*

Rabies virus inoculation in mice.

2or researc".

a* Lao#ato#!animals/

1ultiation of i#uses
http://images.google.com/imgres?imgurl=http://mrshenri.files.wordpress.com/2009/07/chimpanzee-head.jpg&imgrefurl=http://mrshenri.wordpress.com/2009/07/19/banana-test/&usg=__FBG4kgBvX0kam4NrM_SKfSmmnZk=&h=1001&w=1001&sz=271&hl=en&start=5&itbs=1&tbnid=tA5fc1q4e6LLZM:&tbnh=149&tbnw=149&prev=/images%3Fq%3Dchimpanzee%26hl%3Den%26tbs%3Disch:1http://images.google.com/imgres?imgurl=http://mrshenri.files.wordpress.com/2009/07/chimpanzee-head.jpg&imgrefurl=http://mrshenri.wordpress.com/2009/07/19/banana-test/&usg=__FBG4kgBvX0kam4NrM_SKfSmmnZk=&h=1001&w=1001&sz=271&hl=en&start=5&itbs=1&tbnid=tA5fc1q4e6LLZM:&tbnh=149&tbnw=149&prev=/images%3Fq%3Dchimpanzee%26hl%3Den%26tbs%3Disch:1

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* Em#!onate" eggs

http://bigfig%28%273479113013.jpg%27%2C%20%27../fig/big/3479113013.jpg','3479113013.jpg');

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c* 1ell cultu#e (tissue cultu#e


"e most i#ely use#met"o#

issues / t#!%sin separate cells

6ells / g#o4th me"ia in

0at-si#e# bottles

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Virus inoculation 7incubation at 37C (CO2incubators)

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Primary cell

lines

Diploid cell

lines

Continuous cell

lines

Preparedfrom

Organfragments

Human fibroblastseri!e from

embr"onic tissuesTumor cells

Examples #on$e" $ine"Human embr"o lung

tissueHe%a cells from

carcinoma of cer!i&

Number of

Passages(subcultures

'- '- *nlimite

!ypes of cell lines

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etection of Vi#us Re%lication inTissue 1ultu#e/

)- 1!to%athic Eect (1'E/i- 1ell "eath o# l!sis

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ii- .!nc!tial fo#mation (multinucleate#giant cells

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2- T#ansfo#mation/

Viral nucleic aci# / cellular DNA 7 celltrans'ormation 7 'oci o' malignant cells.

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3- Inclusion o"ies

%ite o' i#us asseml! or "egene#atie

changes"eir location an# appearance are

"iagnostic'or a particular virus.

a-Int#ac!to%lasmic/e*g* Raies (Neg#i

o"ies

-Int#anuclea#/e*g.

e#%es i#usesc-+oth/e*g* $easles i#us an" 1$V

7 'l f ti

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7- 'laBue fo#mation/

Infecte" monola!e# C ital "!e 7 unstaine#

areas (%laBues

: aema"so#%tion/

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:- aema"so#%tion/

Monolayer / "emagglutinating virus / R46s7

"ema#sorption (R46sclumping to in'ecte# cells

; I f h

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;* Inte#fe#ence %henomenon/

Monolayer / rubella virus 7 no c"ange 'or

ee!s A## 6*)-pro#ucing virus 7 N56*)

(#ue to

inter'erence>- etection of i#al antigens

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D- i#ect =uo#escent antio"! staining ofinfecte" cells (9A/

"#Neut#ali

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iagnosis of i#alinfections

A- i#ectmetho"s

+- In"i#ect metho"

I* i#ect"etection ofi#uses an" 8

o# thei#com%onents

$$. $solationo' viruses

I*.e#olog

!

$$. %!intests

"i h "

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I* .e#ological "iagnosis/ Detection o' antii#al antio"ies

8 serum samplesacute p"ase & 8-9 ee!s later,to #emonstrate a #ising tite#(: 'ol# increase or more is #iagnostic.

5nly one sample may be use# in t"eacute stage to #etectIg$

+- In"i#ect metho"s

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I* .e#ological "iagnosis/

%erological met"o#s inclu#e

Neutraliation test 6omplement fxation test +aemagglutination in"ibition test $n#irect $2 )$%A

R$A

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II* .@in tests

;se# as an in#ication o' cell-me#iate#immunity (6M$ in some viral in'ections, e.g.mumps.

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M6

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)Vi#uses "ie# f#om acte#ia inall of the follo4ing E1E'T/

a Viruses are very small in sie.b Viruses are obligatory intracellular

parasites.

c Viruses contain bot" DNA an# RNA.# Viruses cannot be cultivate# on

artifcial culture me#ia.

e Viruses are not susceptible toantibiotics.

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2 .ites of i#al asseml! in tissue

cultu#e a#e calle"/a*la1ues

b$nclusion bo#ies

cViral capsi##6*)

eAreas o' trans'ormation

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3 Rega#"ing continuous cell lines6 allof the follo4ing a#e t#ue E1E'T/

a"ey allo unlimite# number o'passages.

b"ey are prepare# 'rom tumor cells.cA'ter viral inoculation, t"ey s"oul# be

sterilie# by autoclaving.

#"ey are use# 'or isolation o' viruses.

e+ea cell line is an example o'continuous cell lines.

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7 '1R ma! e use" to "iagnosei#al infections ! "etecting/

aAntiviral $gM antibo#ies

bViral antigens

cRising titer o' antiviral antibo#ies

#Viral nucleic aci#

e6*) pro#uce# by t"e virus in tissue

culture

: i " i f i

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: i#ect "etection of i#usesan"8o# thei# com%onents can e

"one ! all of the follo4ingE1E'T/

a2luorescent microscope

b)lectron microscopecig"t microscope

#*6R

e4ioc"emical reactions

; 1 lti ti f i

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; 1ultiation of i#uses can e"one on/

a4loo# agarbissue culture

cMac6on!ey=s me#ium

#Nutrient brot"eAnaerobic me#ia

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TANF

YO?

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)Vi#uses "ie# f#om acte#ia in all

the follo4ing E1E'T/a Viruses are very small in sie.

b Viruses are obligatory intracellular

parasites.c Viruses contain bot" DNA an# RNA.

# Viruses cannot be cultivate# on

artifcial culture me#ia.e Viruses are not susceptible to

antibiotics.

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2 .ites of i#al asseml! in tissue

cultu#e a#e calle"/a*la1ues

b$nclusion bo#ies

cViral capsi##6*)

eAreas o' trans'ormation

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3 Rega#"ing continuous cell lines6all of the follo4ing a#e t#ue

E1E'T/a"ey allo unlimite# number o'

passages.

b"ey are prepare# 'rom tumor cells.cA'ter viral inoculation, t"ey s"oul# be

sterilie# by autoclaving.

#"ey are use# 'or isolation o' viruses.e+ea cell line is an example o'

continuous cell lines.

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7 '1R ma! e use" to "iagnosei#al infections ! "etecting/

aAntiviral $gM antibo#ies

bViral antigenscRising titer o' antiviral antibo#ies

#Viral nucleic aci#

e6*) pro#uce# by t"e virus in tissueculture

: i#ect "etection of i# ses

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: i#ect "etection of i#usesan"8o# thei# com%onents can e

"one ! all of the follo4ingE1E'T/

a2luorescent microscope

b)lectron microscopecig"t microscope

#*6R

e4ioc"emical reactions

; 1ultiation of i#uses can e

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; 1ultiation of i#uses can e"one on/

a4loo# agarbissue culture

cMac6on!ey=s me#ium

#Nutrient brot"eAnaerobic me#ia

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