IDENTIFICATION

OF VIRUS IN
CELL CULTURE
 WHY CELL CULTURE IS REQUIRED FOR
VIRUSES? WHY NOT MEDIA CULTURE?

• THE GROWTH OF VIRUSES REQUIRES CELL CULTURE BECAUSE

VIRUSES REPLICATE ONLY IN LIVING CELLS, NOT ON CELL-FREE
MEDIA THE WAY MOST BACTERIA CAN.
• VIRUSES ARE INACTIVATED AT ROOM TEMPERATURE, SO IT IS
IMPORTANT TO INOCULATE THE SPECIMEN INTO THE CELL
CULTURE AS SOON AS POSSIBLE.
 CHARACTERISTIC CYTOPATHIC EFFECT
(CPE)
• VIRUS GROWTH IN CELL CULTURE FREQUENTLY PRODUCES A
CHARACTERISTIC CYTOPATHIC EFFECT (CPE) THAT CAN PROVIDE
A PRESUMPTIVE IDENTIFICATION.
• CPE IS A CHANGE IN THE APPEARANCE OF THE VIRUS-INFECTED
CELLS. THIS CHANGE CAN BE IN SUCH FEATURES AS SIZE, SHAPE,
AND THE FUSION OF CELLS TO FORM MULTINUCLEATED GIANT
CELLS (SYNCYTIA).
• CPE IS USUALLY A MANIFESTATION OF VIRUS-INFECTED CELLS
THAT ARE DYING OR DEAD.
 IF THE VIRUS DOES NOT PRODUCE A CPE, ITS
PRESENCE CAN BE DETECTED BY SEVERAL
OTHER
TECHNIQUES:
1. HEMADSORPTION (ATTACHMENT OF ERYTHROCYTES TO THE
SURFACE OF VIRUS-INFECTED CELLS.
2. INTERFERENCE WITH THE FORMATION OF A CPE BY A SECOND
VIRUS.
3. DECREASE IN ACID PRODUCTION BY INFECTED AND DYING CELLS
 A DEFINITIVE IDENTIFICATION OF THE
VIRUS GROWN IN CELL CULTURE IS MADE BY
USING
KNOWN ANTIBODY IN ONE OF SEVERAL
1. COMPLEMENT FIXATION
TESTS:
2. HEMAGGLUTINATION INHIBITION
3. NEUTRALIZATION
4. FLUORESCENT ANTIBODY ASSAY
5. RADIOIMMUNOASSAY
6. ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA)
7. IMMUNOELECTRON MICROSCOPY
THANK YOU