PRACTICAL LESSON 22

THEME: ENTEROVIRUSES. RABDOVIRUSES. LABORATORY DIAGNOSTICS OF

POLYOMYELITIS AND RABIES.
Theme’s actuality. Poliomyelit is viruses, Coxsackie, ECHO, 68-72 types of enteroviruses spread
by type of intestinal infections and play an important role in human pathology, causing damage to
the central nervous system, muscular and skeletal system, and liver. As a result of mass vaccination,
the previously widespread poliovirus is now an epidemic disease. However, the pathogen continues
to circulate along with the vaccine strain, which can lead to an exacerbation of the epidemic danger.
Rabies from antiquity aroused fear in people and resulted death. Despite the existing measures to
prevent this dangerous disease, it remains relevant in connection with the spread of stray animals on
the streets of cities. The modern doctor needs not only to be able to distinguish between viral
diseases from diseases caused by microbial pathogens, but correctly prescribe the treatment and be
able to carry out the prevention of viral diseases.

General purpose – to be able to conduct virological diagnosis of polyomyelitis, Coxsackie, ECHO

and rabies, and also use the necessary drugs to specifically prevent the corresponding infections.

Specific purposes – to know how:

1. To identify the detected enterovirus in the neutralization reaction.
2. To analyze the features of the biological method of diagnosis for various enterovirus infections.
3. To identify the rabies virus through intracellular inclusions.
Basic level of knowledge, skills:
1. Analyze the features of the interaction of viruses with living systems.
2. Explain the features of the immune response in viral infections.
3. Treat the biological properties of viruses as a special non-cellular form of life.
4. Describe the differences in the structure of the viral cell from the microbial.

We offer you few tasks to test your level of knowlege:

Task 1. What are the differences between simple and complex viruses?
Task 2. What are the consequences for the cell has a productive viral infection?
Task 3. How to identify CPD in the culture of infected cells?
Task 4. What components are needed for the viral neutralization reaction?
Task 5. Rabies refers to zoonotic infections. What does it mean?
Task 6. Enteroviruses and rhabdoviruses are RNA genomic. However, the virion of rhabdoviruses
contains the enzyme RNA-dependent RNA polymerase, and the enterovirus virion does not. How
can this be explained?

Information for improving your knowledge and skills can be found in following books:
1. Medical microbiology, virology and immunology /redacted by L.B.Borisov.-Medical Information
Agency.-Moscow.-2002.-pages 68-90.
1.Virology / redacted by А. G. Bukrinska: Volume 2, Virtual reality Ltd , 2012, pages11-151
2. Medical microbiology, virology and immunology / redacted byBorisov L.B., SmirnovaA.M..- М
.: Medicine, 2005. –pages30-45, 328-337.
4.AfanasyevY.I., YurinaN.А. Histology, cytology and embryology.- М .: Medicine.- 2002.- pages
42-75.
5. . Medical microbiology, virology and immunology: Textbook / redacted by. V.P. Shirobokov.–
Vinnitsia : „Novaya Kniga”, 2015. – pages280-288; 299-302.
6. Vorobyov А.А. et al. Microbiology.- М.: Medical Information Agency , 2012.- pages312-316;.
323-324; 587, 525, 538.
7. Clinical laboratory analytics in five volumes/Volume IV – Clinical microbiology/ redacted by
V.V. Menshikov. - М.:Agat-Мed, 2003.-pages623-628, 664.
Theoretical questions on the basis of which is possible the implementation of the targeted
activities.
1.Viruses that cause viral gastroenteritis. General properties of thegroups, role in human pathology.
2. Rhinoviruses as the main pathogens of viral gastroenteritis. Methods of virological diagnosis.
3. Family of picornaviruses. General properties, family composition.
4. The breed of enteroviruses. General properties of the genus. Types, their classification,
distribution, role in pathology.
5. The driver of poliomyelitis. Biological properties, serotypes. Epidemiology and specific
prevention.
6.Viruses Coxsackie and ECHO – representatives of enteroviruses, their classification, distribution,
role in pathology.
7. Clinical forms of diseases caused by Coxsackie A and B viruses, ECHO viruses, serotypes 70
and 71. Virological diagnosis.
8. The genus of aptoviruses. Viruses of foot and mouth disease. Biological properties.
Classification, pathogenesis of infection in humans, specific prophylactics.
9. The rabies virus. General characteristics. Role in human pathology.
10. The virus of vesicular stomatitis. Biological properties.
Literature:
1. Borisov L.B.Medical microbiology, virology and immunology. – М., МIА, 2005.- pages 485-490,
527-529.
2. Korotyaev А.I., Babichev S.А. Medical microbiology, virology and immunology: Textbook for
medical schools – 4th edition, revised and enlarged.-SPb.: SpetsLit, 2008. –pages347-352, 387-
390.
3. Medical microbiology, virology and immunology: Textbook / redacted by. V.P. Shirobokov.–
Vinnitsia : „Novaya Kniga”, 2015. – pages491-495, 549-554.
4. Microbiology and immunology for dentists/ redacted by. R. J.Lamont, R. A. Burne, , M. S. Lantz,
& D. J. LeBlanc. –М.:Practical medicine, -2010.- pages352.
5. .Virology / redacted by А. G. Bukrinska: Volume 2, Virtual reality Ltd , 2012, pages11-151,
210, 253.
6. Vorobyov А.А. et al. Microbiology.- М.: Medical Information Agency , 2012.- pages312-316;.
323-324; 587, 525, 538, 569.
7. Clinical laboratory analytics in five volumes/Volume IV – Clinical microbiology/ redacted by
V.V. Menshikov. - М.: Agat-Мed, 2003.-pages623-628, 664
8. Medical microbiology, virology and immunology: Textbook / redacted by. V.P. Shirobokov.–
Vinnitsia : „Novaya Kniga”, 2015. – pages442,495.

Materials and equipment: Virological mattresses with a demonstration of CPD, neutralization

reaction, tables and biologics on the subject, microscopic preparations of the brain affected by the
rabies virus.

An indicative basis for action in the performance of practical work

1. Conduct an indication of enteroviruses according to the CPR on the culture of HeLa cells. To
consider and compare micro-preparations of uninfected culture and infected pathological material
from patients No. 1 and No. 2. Pay attention to the change in the morphology of cells under the
influence of the virus. Draw the pictures in the protocol, write a conclusion about the etiology of the
disease.
2.Carry out an indication of the detected enterovirus in the neutralization reaction. To do this, insert
tubes containing the culture of FBS cells (human embryo fibroblasts) to make the material
containing the virus, mix in git with the standard polyvalent serum against the viruses of the
poliomyelitis group, Coxsackie or ECHO. In the case of a coincidence of the type of virus and
serum, the virus is neutralized and it does not cause CPA in the cell culture. Note the results in the
protocol, write a conclusion about the species belong to the detected virus.
3. Using micro preparations of brain sections of a person who died from rabies and the table
"Rhabdoviruses" get acquainted with the microscopic method of diagnosis of rabies. Draw the
micro preparation, note intracellular inclusions (Babes-Negri bodies).

4.From the set of immunobiological drug choose drugs that are used to prevent enterovirus
infections and rabies.
 Addition №1
Graphic structure of the theme «Enteroviruses. Rhabdoviruses. Laboratory diagnostics of
poliomyelitis and rabies »

Enteroviruses

Classification Methods of diagnostics

Biological test Virological

(infection of mice)
Poliomyelitis

Coxsackie
ЕСНО

Pathologic material

Invasion of cell
Biological properties culture

CPA
Features of Resistance to
Virion structure
reproduction various factors

Identification
Ways of (neutralization
Antigens Cultivation transmission of reaction with
infection specific sera,
RSK)
Serologic
diagnostics Nonspecific
prevention

Specific
prophylaxis
 Graphic structure of the theme «Enteroviruses. Rhabdoviruses. Laboratory diagnostics of
poliomyelitis and rabies »

RHABDOVIRUSES

Biological properties Methods of

diagnostics

Virological
Virion structure
Natural source

Antigens Biological test

Ways of
transmission of
infection

Specific
prophylaxis Viroscopic

The history of the

vaccine’s creation
 Addition №2
Sample protocol for the practical lesson «Enteroviruses. Rhabdoviruses. Laboratory
diagnostics of poliomyelitis and rabies»

Task 1.Enterovirus indication using cytopathic

action.

Picture 1.Uninfected cell culture Picture 2.Cell culture infected Picture 3.Cell culture infected
(control) with patch material from with patch material from
patient#1 patient #2
Conclusion: __________________________________________________________________

Task 2. Enterovirus indication using neutralization reaction.

Standard multivalent Group of Coxsackie Group ЕСНО Control

sera poliomyelitis Group
Reaction

„+” – CPA, „-” - lack of CPA

Conclusion:__________________________________________________________

Task 3. Microscopic diagnosis of rabies.

Picture 4. Intracellular inclusions (Babes-Negri bodies)

Task 4. Preparations for specific prevention of enterovirus and rhabdovirus infections

Date _____________ Lecturer’s signature ____________