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CONTENTS
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INTRODUCTION
CLASSIFICATION OF VIRUSES
Herpes simplex infections
Herpangina
Acute lymhonodular pharyngitis
Hand ,foot & mouth disease
Measles
Rubella
Chickenpox
Herpes zoster
Small pox
Mumps
Infectious mononucleosis
Cytomegalovirus
HIV
Hepatitis
Laboratory diagnosis of viral infections
Conclusion
References
INTRODUCTION
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Infinite in distribution.
Occupy the twilight zone that separates the living from non living
Viruses offer the best models for understanding the chemistry of life
CLASSIFICATION
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RNA
DNA
RNA viruses
DNA viruses
RNA VIRUSES
1) orthomyxovirus
influenza
2)Paramyxovirus
3)Rhabdovirus
4)Arenavirus
Measles, mumps
Rabies, hemorrhagic fever
Lymphocytic choriomeningitis, lassa
fever
5)calcivirus
6)Coronavirus
7)Bunyavirus
8)picornavirus
Poliomyelitis, coxsackie
diseases,commoncold, foot and mouth
disease
9)Reovirus
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10) Togavirus
11)Retrovirus( RNA tumor virus
DNA VIRUSES
1) Herpes virus
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a)Herpes simplex 1
Gingivostomatitis, keratconjunctivitis,
genital and skin lesions
b)Herpes simplex 2
d)cytomegalovirus
Infectious mononucleosis
Roseola infantum
Infectious mononucleosis
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2)Pox virus
3)adenovirus
Pharyngoconjunctival fever ,
epidemic keratoconjunctivitis
4)parvovirus
5)iridovirus
6)Papovirus
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3) Latent infections
Herpes simplex and varicella zoster viruses remain
latent in the nerve root ganglion, to be reactivated
periodically in some individuals causing recurrent
lesions
Hepatitis B virus chronic infections which may remain
inapparent for many years
Subacute sclerosing panencephalitis- develops between
1- 10 years after recovery of measles virus infection
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Chickenpox
Herpes zoster
Small pox
Mumps
Infectious mononucleosis
Cytomegalovirus
HIV
Hepatitis
HERPES SIMPLEX
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PATHOGENESIS
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FORMS OF HSV
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Herpes genitalis
HSV -2
Uterine cervix,vagina,vulva & penis.
Now often termed as new epidemic venereal diseasesince it is transmitted through sexual contact
More virulent than HSV-1
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Herpes meningoencephalitis
Sudden fever & symptoms of increased intracranial pressure.
Paralysis of muscle groups, convulsions and death may occur
Herpes conjunctivitis /Keratoconjunctivitis
Swelling & congestion of palpebral conjunctiva.
Keratitis & corneal ulceration.
Herpetic vesicles of eyelids are seen which heal rapidly.
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It rarely occurs below the age of six months--because of presence of circulating antibodies in the
infant derived from the mother
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Ulcers vary considerably in size - from tiny lesions to measuring several millimeters to even a centimeter in
diameter
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HISTOLOGICAL FEATURES
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intranuclear inclusions
Lipschutz bodies
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LABORATORY DIAGNOSIS
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Cytology smears
A smear taken of epithelial cells at the base of a suspected lesion
studied to determine if epithelial cells show changes consistent
with HSV infection.
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Tzanck test
Tzanck preparation is a rapid test done to diagnose
infections caused by herpesviruses.
Cells are examined under a microscope for signs of
infection
The Tzanck preparation is done by smearing cells taken
from a fresh blister or ulcer onto a microscope slide.
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The cells are stained with a special stain, such as Wright's stain,
and then examined under a microscope for characteristic
changes caused by a herpesvirus
Herpes causes giant cells with multiple nuclei
The background of the cell looks like ground glass and contains
small dark spots called inclusion bodies
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Immunomorphologic tests
The positively infected cells are fluorescent green when examined under a
fluorescent microscope.
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Serologic tests
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TREATMENT
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LABORATORY DIAGNOSIS
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Treatment
Acyclovir
Vidarbine
Idoxuridine
HERPANGINA
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CLINICAL FEATURES
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LABORATORY FINDINGS
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Treatment
No treatment is necessary as disease - self limiting
& have few complications
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ORAL MANIFESTATIONS
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LABORATORY FINDINGS
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Histologic features
Papules or nodules consist of hyperplastic
lymphoid aggregates.
In some cases, intranuclear inclusion bodies are
seen.
Treatment
No treatment is necessary as disease is self
limiting
Etiology Coxsackie A 16
Less frequently by A5 A6
Appearance of maculopapular ,
exanthematous & vesicular lesions of skin
particularly involving the hands, feet, legs
and arms
ORAL MANIFESTATIONS
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LABORATORY FINDINGS
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Clinical features
MEASLES
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Rubeola,Morbilli.
Acute, contagious, dermatropic viral infection
primarily affecting children
Etiology - Paramyxovirus (RNA virus)
Incubation period - 8-12 days
Transmission - respiratory secretions or direct
contact of droplets.
PATHOGENESIS
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CLINICAL FEATURES
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HISTOPATHOLOGICAL FEATURES
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CONTROL MEASURES
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RUBELLA/GERMAN MEASLES
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CLINICAL FEATURES
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ORAL MANIFESTATIONS
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DIAGNOSIS
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1)small pox eradication has been achieved through out the world
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CLINICAL FEATURES
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Papules
Vesicles
Pustules
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MOLLUSCUM CONTAGIOSUM
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CLINICAL FEATURES
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HISTOPATHOLOGICAL FEATURES
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CONDYLOMA ACUMINATUM
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CLINICAL FEATURES
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HISTOPATHOLOGICAL FEATURES
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CHICKENPOX(VARICELLA)
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CLINICAL FEATURES
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ORAL MANIFESTATIONS
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Complications
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HERPES ZOSTER
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Shingles/Zona
Acute infectious viral disease - extremely painful and
incapacitating nature - characterized by inflammation of
dorsal root ganglia or extra- medullary cranial nerve ganglia
CLINICAL FEATURES
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ORAL MANIFESTATIONS
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Diagnosis:
Recognized by its characteristic distribution of lesions
Skin & oral lesions identified by cytologic smear &
finding of Tzanck cells.
To differentiate between herpes zoster and simplex--fluroscent antibody staining technique, seroliogical
diagnosis and viral culture can be done
MUMPS
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Epidemic parotitis
Transmission -
respiratory route.
PATHOGENESIS
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CLINICAL FEATURES
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Firm, rubbery or elastic swelling of the salivary glands elevating the ear -lasts for one week.
3 days
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DIAGNOSIS
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COMPLICATIONS
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Prevention
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MMR vaccine
1st dose 9 months
2nd dose- 15 -18 months
INFECTIOUS MONONUCLEOSIS
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CLINICAL FEATURES
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Body temperature may reach 104F and lasts from 2-14 days
ORAL MANIFESTATIONS
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LABORATORY FINDINGS
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White blood cells count is increased, with the differential count showing
relative lympocytosis
Oval, horse shoe shaped or indented nuclei with dense irregular nuclear
chromatin and a basophilic foamy cytoplasm
PROCUDURE
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EBV-SPECIFIC ANTIBODIES
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CYTOMEGALOVIRUS
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HISLOPATHOLOGIC FEATURES
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DIAGNOSIS
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ACQUIRED IMMUNODEFICIENCY
SYNDROME (AIDS)
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Etiology
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HIV-2 -
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Vishakhapatnam
East Godavari
Warangal
Medak
Chittoor
Kurnool
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5
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Stage 1 - Primary
Stage 2 Asymptomatic
Stage 3 - Symptomatic
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CLINICAL FEATURES
AIDS
- diagnosed
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Bacterial infections
Mycobacterium avium intercellulare,
Mycobacterium tuberculosis
Melanotic hyperpigmentation
Necrotizing ulcerative stomatitis
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Group 3:
Bacterial infections
Actinomyces israelii
Escherichia coli
Klebsiella pneumoniae
Cat-scratch disease
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Neurologic disturbances
Facial palsy,Trigeminal neuralgia
Recurrent apthous stomatitis
Viral infections CMV, Molluscum contagiosum
ORAL CANDIDIASIS
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Pseudomembranous candidiasis
White to yellowish white plaques - easily scraped off,
exposing red areas.
Lesions - extensive, involving > one site in the oral
cavity.
Involve the oropharynx and eosophagus
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Erythematous candidiasis
Clinically seen as red lesions - located on the dorsum
of the tongue, palate, and buccal mucosa.
Tongue lesions are also referred to as central papillary
atrophy.
HYPERPLASTIC CANDIDIASIS
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ANGULAR CHEILITIS
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PERIODONTAL LESIONS
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Is a non-plaque induced
gingivitis.
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Subclassified as
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NECROTIZING STOMATITIS
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HERPES ZOSTER
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imparting a corrugated
appearance.
KAPOSI'S SARCOMA
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APHTHOUS ULCERS
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MOLLUSCUM CONTAGIOSUM
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NON-HODGKIN'S LYMPHOMA
(NHL)
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HYPERPIGMENTATION
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SPECIFIC TESTS
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Virus isolation
Antigen detection
Antibody detection ELISA
Western blot test
Polymerase chain reaction
VIRUS ISOLATION
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PROCEDURE
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USES
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Disadvantage
Not suitable as a routine diagnostic test.
Risky
With the availability of PCR, HIV culture is not done routinely.
ANTIGEN DETECTION
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SANDWICH ELISA
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USES
persons recently exposed to risk of infection and in
whom antibody test is negative.
Infants born of seropositive mothers.
screening blood donors
2 TYPES
SCREENING TESTS,
SUPPLEMENTAL OR CONFIRMATORY
TESTS.
SCREENING TESTS
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ELISA
Principle
viral antigen is coated on surface of
microwells
Main requirements
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1.Serum or plasma
2.Diagnostic kit
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Procedure
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Interpretation:
Positive result - colour
Negative result - no colour
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USE
Posses high sensitivity
Have a broadly reactive spectrum
Simple to perform
Can be automated for handling large sample at a time
DISADVANTAGES
Not highly specific
May give a few false positive results
It is inconvenient for testing single sample quickly.
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SALIVA
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prognostic indicator
KARPAS TEST
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Fluorochrom
e
Labeled
Anti-Ig
Unlabeled
Ab
Ag
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SUPPLEMENTAL TEST
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Main requirements
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1) serum(or plasma)
2) Diagnostic kit:
a) Nitrocellulose strips activated by the transfer of
HIV-1 viral proteins.
b) Conjugate: Alkaline phosphatase(AP)-labelled
antihuman IgG goat antibodies.
c)
Buffered-substrate: Nitro-blue Tetrazolium
(NBT).
d) Diluent solution(wash solution).
Procedure
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Disadvantage
Interpretation remains subjective & demands
considerable experience
Indeterminate results
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Indeterminate WB test
Recent HIV infections.
Advanced immunodeficiency because of loss of
antibodies.
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Two types
1)PCR for DNA of provirus present in the infected
host cell.
2) RT-PCR for HIV RNA, which detects free virus
present in the plasma.
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REQUIREMENTS
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PROCEDURE
3 steps
Denaturation
Annealing
Extension
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Denaturation--- DNA is
separated into two
strands
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Extension
DNA polymerase & raw
materials in the form of
nucleotides are added
Beginning at the free
ends of the primers,
polymerase extend the
molecule by adding
appropriate nucleotides
Uses:
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Limitations :
its cost
Need for sophisticated laboratory equipment
Are indicated only when other methods cannot
give a definitive results
ELISA
antibodie
s
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Positive
Repeat ELISA
Positive
Positive
HIV infection
Positive
confirm
with WB
indeterminate result
suspect HIV
PCR for
HIV
Negative
infection
negative
Not HIV
Flow cytometry
Measurements are made while cells in a liquid
suspension are forced to flow one at a time
through a measuring device.
Method
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Uses
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Specimen collection
Cultivation of viruses
Virus isolation
Microscopy
Demonstration of virus antigen
Demonstration of antibodies
SPECIMEN COLLECTION
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CULTIVATION OF VIRUSES
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1. Animal inoculation:
Infant suckling mice are the most widely employed
animals in virology
Infant mice are used in the isolation of coxsackie and
arboviruses
Mice may be inoculated by several routes
intracerebral, subcutaneous, intraperitoneal or
intranasal.
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Uses
Primary isolation of certain virus
Used for study of pathogenesis of viral diseases,
To study viral oncogenesis.
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Chorioallantoic membrane(CAM)
Mainly for growing pox virus- produces visible lesions called
(pocks).
Each infectious virus particle can form one pock.
Pock counting there fore indicates the no of viruses in the
inoculum
Different viruses produce pocks of different morphology
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TISSUE CULTURE
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Explant culture
Fragments of minced tissue can be grown as
explants embedded in plasma clots. They may
also be cultivated in suspension.
This method is now seldom employed in
virology.
CELL CULTURE
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These are cells of a single type that contain the same number of chromosomes as
the parent cells
They can be subcultured for a limited number
After about 50 serial subcultures they undergo senescence and the cell strain is
lost
DIPLOID cells developed from the human fibroblasts are susceptible to a number
of human viruses
cytopathic effect:
many viruses cause morphological changes in cultured cells in which they grow
these changes are known as cytopathic effects (cpe) viruses causing cpe --cytopathogenic viruses
syncytium formation
cellularclumping
rounding of cells
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HAEMADSORPTION
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INTERFERENCE
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METHOD
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Conclusion
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REFERENCES
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Thank
you
THANK YOU
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Eclipse phase
From the stage of penetration of virus
into the host cell till the appearance of
first infectious virus progeny particle, the
virus cannot be demonstrated inside the
host cell
This period is known as eclipse phase
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Droplet nuclei
Droplet nuclei droplets of size less than
5m which cannot travel more than 3
feet
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ELVIS
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Principle of PCR
minute quantities of specific DNA or RNA
sequence can be enzymatically amplified
to the extent that a sufficient quantity of
material is available for detection
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Herpes barbae
Herpes simplex spread over the
bearded region of the face into the minor
injuries created by daily shaving leading
to condition known as herpes barbae
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Koplik spots
Immune reaction to the virus in the
endothelial cells of dermal capillaries
plays a role in the development of spots
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Cell tropism
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TB in AIDS
Caused by Mycobacterium. Avium and M.
intracellulare , M. bovis, M.
scrofulaceum, M. africanum and M.
haemophilum usually found in
immunocompromised patients
Oral lesions are uncommon, when
present tongue is frequently
Affected areas --- chronic ulcerations or
exophytic proliferative masses
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TRIDOT
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Principle:
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Cost effective
Antibodies directed against a specific
antigen can be produced invitro by
hybridoma technique