PICORNAVIRUSES

(Enterovirus & Rhinovirus groups)

Introduction

Picornaviruses a very large virus family with respect to the number of members

But smallest in terms of virion size and complexity.

Enteroviruses are alimentary tract

transient

inhabitants

of

human

May be isolated from throat or lower intestine.

Rhinoviruses are isolated from nose and throat

Introduction contd

Replication occur in cytoplasm of host cells Cause diseases in humans ranging from severe paralysis to aseptic meningitis

Pleurodynia (Bornholm Disease), exanthematous skin lesions,

myocarditis,

vesicular

and

Mucocutaneous lesions, respiratory illnesses, Undifferentiated febrile illness, generalized disease of infant conjunctivitis and severe

Properties & Classification of Picornaviruses

Virion is having icosahedral symmetry, 24-30nm. It contain single stranded positive sense RNA. There is no envelope Contains six genera:

Enterovirus (enteroviruses) Rhinovirus(rhinoviruses) Hepatovirus (hepatitis A virus) Parechovirus (parechoviruses), Aphthovirus (foot-and-mouth disease viruses) Cardiovirus (Cardioviruses)

The first four groups contain important human pathogens.

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Enteroviruses Group

Enteroviruses

Enteroviruses are a genus of the picornavirus family which replicate mainly in the gut Single stranded naked RNA virus with icosahedral symmetry Unlike rhinoviruses, they are stable in acid pH Capsid has 60 copies each of 4 proteins, VP1, VP2, VP3 and VP4 arranged with icosahedral symmetry around a positive sense genome.
At least 72 serotypes are known: divided into 5 groups

Polioviruses Coxsackie A viruses Coxsackie B viruses Echoviruses type Enteroviruses (more recently, new enteroviruses subtype have been allocated sequential numbers (68-72))
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Enterovirus Particles

Courtesy of Linda M. Stannard, University of Cape Town, S.A.h

Properties of Enteroviruses
Group Virus types 3 types (1 - 3) CPE in cell cultures Monkey Human cell kidney culture + + - or I Pathology in newborn mice + Major disease associations Paralytic poliomyelitis, aseptic meningitis, febrile illness. Aseptic meningitis, herpangina, febrile illness, conjunctivitis (A24), hand, foot and mouth disease. Aseptic meningitis, severe neonatal disease, myopericarditis, Bornholm disease, encephalitis, febrile illness. Aseptic meningitis, rash, febrile illness, conjunctivitis, severe generalized neonatal disease. Polio-like illness, aseptic meningitis, hand, foot and mouth (E71), epidemic conjunctivitis (E70) hepatitis A (E72) Poliovirus Coxsackie group A Coxsackie group B

23 types - or I (A1-22, A24) 6 types (B1-6) +

Echovirus

31 types (1-9, 11-27 29-33) 5 types (68-72)

Enterovirus

Poliovirus

Three serotypes of poliovirus (1, 2, and 3) but no common antigen. Have identical physical properties but only share 36-52% nucleotide homology. Humans are the only susceptible hosts Polioviruses are distributed globally

The availability of immunization and the poliovirus eradication campaign has eradicated poliovirus in most regions of the world except in the Indian Subcontinent and Africa.
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Pathogenesis

The incubation period is usually 7 - 14 days. Following ingestion, the virus multiplies in the oropharyngeal and intestinal mucosa.

The lymphatic system, in particular the tonsils and the Peyer's patches of the ileum are invaded and the virus enters the blood resulting in a transient viraemia In a minority of cases,the virus may involve the CNS following dissemination.

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Clinical Manifestations
There are 3 possible outcomes of infection:

Subclinical infection (90 - 95%) - inapparent subclinical infection account for the vast majority of poliovirus infections Abortive infection (4 - 8%) - a minor influenza-like illness occurs, recovery occurs within a few days and the diagnosis can only be made by the laboratory

The minor illness may be accompanied by aseptic meningitis

Major illness (1 - 2%) (poliomyelitis) - the major illness may present 2 - 3 days following the minor illness or without any preceding minor illness.

Signs of aseptic meningitis are common. Involvement of the anterior horn cells lead to flaccid paralysis. Involvement of the medulla may lead to respiratory paralysis and death.
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Laboratory Diagnosis

Virus Isolation

Mainstay of diagnosis of poliovirus infection Can be readily isolated from throat swabs, faeces, and rectal swabs. It is rarely isolated from the CSF Can be readily grown and identified in cell culture Requires molecular techniques to differentiate between the wild type and the vaccine type. Very rarely used for diagnosis since cell culture is efficient. Occasionally used for immune status screening for immunocompromised individuals.
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Serology

Epidemiology

Children are most susceptible Humans are only reservoir of infection Crowded conditions, poor hygiene and sanitation favors the transmission It is mainly transmitted by contaminated water with drinking water

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Prevention contd..
No specific antiviral therapy is available However, the disease may be prevented through vaccination There are two vaccines available Intramuscular Poliovirus Vaccine (IPV)

Consists of formalin inactivated virus of all 3 poliovirus serotypes Produces serum antibodies only: does not induce local immunity and thus will not prevent local infection of the gut However, it will prevent paralytic poliomyelitis since viraemia is essential for the pathogenesis of the disease.
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Prevention contd..

Oral Poliovirus Vaccine (OPV)

Consists of live attenuated virus of all 3 serotypes Produces local immunity through the induction of an IgA response as well as systemic immunity. Rarely causes paralytic poliomyelitis, around 1 in 3 million doses.

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Prevention contd.

Most countries use OPV because of its ability to induce local immunity The normal response rate to OPV is close to 100%. OPV is used for the WHO poliovirus eradication campaign. Poliovirus was targeted for eradication by the WHO by the end of year 2000

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Coxsackieviruses

Coxsackieviruses are distinguished from other enteroviruses by their pathogenicity for suckling rather than adult mice
They are divided into 2 groups on the basis of the lesions observed in suckling mice.

Group A produce a variety of illness like herpangina (vesicular pharyngitis), hand-foot-and-mouth disease and acute hemorrhagic conjunctivitis. Group B causes pleurodynia (epidemic myalgia), myocarditis, pericarditis, meningoencephalitis and severe generalized disease of children Group A and B may also cause aseptic meningitis, respiratory and undifferentiated febrile illnesses, hepatitis and paralysis Paralysis produced by non polioviruses is incomplete and reversible Coxsackie B viruses are most commonly identified agents of viral heart disease
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Laboratory diagnosis

Virus can be isolated form throat washings, stool, nasal secretions. In aseptic meningitis, CSF In hemorrhagic conjunctivitis virus is isolated from conjunctival swabs Samples are inoculated in tissue cultures and also in suckling mice. Tissue culture show CPE after 5-14 days. Suckling mice show signs of illness appear within 3-8 days (group A) and 5-14 days (group B). Serologic diagnosis can be done by Neutralization , immunofluorescence technique
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Epidemiology

It is encountered all round the world Isolations made from human feces, pharyngeal swabs, sewage and flies

Familial exposure is important in acquiring the infection.

No vaccine or antiviral drugs are available

Control:

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Echoviruses

The first echoviruses were accidentally discovered in human faeces, unassociated with human disease during epidemiological studies of polioviruses.

The viruses were named echoviruses (Enteric, Cytopathic, Human, Orphan viruses).

These viruses were produced CPE in cell cultures, but did not induce detectable pathological lesions in suckling mice. Altogether, there are 32 echoviruses (types 1-34; echovirus 10 and 28 were found to be other viruses and thus the numbers are unused) There is no group echovirus Ag but heterotypic cross-reactions occur between a few pairs.
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New Enteroviruses

Newly identified picornaviruses that are not polioviruses

No longer classified separated into the species coxsackie and echovirus because of the ambiguities presented by overlapping host range variations.

Four new enteroviruses have been identified (68 - 72).

Enterovirus 70 is the causative agent epidemics of acute haemorrhagic conjunctivitis

That swept through Africa, Asia, India and Europe from 1969 to 1974. The virus is occasionally neurovirulent.
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New Enteroviruses contd

Enterovirus 71 appears to be highly pathogenic

Associated with epidemics of a variety of acute diseases Aseptic meningitis, encephalitis, paralytic poliomyelitis-like disease and hand-foot-mouth disease.

Enterovirus 72 was originally assigned to hepatitis A virus,

But it had now been assigned to a new family called heptoviruses

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Diseases summary with Enteroviruses

Syndrome Polio Paralytic disease + Meningitis-encephalitis + Carditis + Neonatal disease Pleurodynia Herpangina Rash disease Haemorr. conjunctivitis Respiratory infections + Undifferentiated fever + Diabetes/pancreatitis Cox A + + + + + + + + Cox B + + + + + + + + + Echo + + + + + + + -

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Laboratory Diagnosis

Virus Isolation

Mainstay of diagnosis of enterovirus infection Coxsackie B and Echoviruses can be readily grown in cell culture from throat swabs, faeces, and rectal swabs. They can also be isolated from the CSF Coxsackie A viruses cannot be easily isolated in cell culture. Molecular techniques may provide a better alternative Very rarely used for diagnosis since cell culture is efficient. Neutralization tests or EIAs are used but are very cumbersome and thus not offered by most diagnostic laboratories

Serology

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Cytopathic Effect

(Virology Laboratory, New-Yale Haven Hospital)

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Treatment and Prevention

There is no specific antiviral therapy available against enteroviruses other than polio. Some authorities use IVIG (Immune globulin) in the treatment of neonatal infections or severe infections in immunocompromised individuals. However, the efficacy is uncertain. HNIG (Human normal immunoglobulin) have been to prevent outbreaks of neonatal infection with good results. There is no vaccine available mainly because of the multiplicity of serotypes. There is little interest in developing a vaccine except against enterovirus 71 and coxsackie B viruses.

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RHINOVIRUS GROUP

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RHINOVIRUS

They are called common cold viruses They are commonly isolated from mild upper respiratory illnesses

They are usually isolated form nasal secretions, throat and oral secretions.

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Characteristics of Rhinovirus

These are picornaviruses similar to enteroviruses

But differ from them in having acid lability

Thermostable than enteroviruses and survive for days

Animal susceptibility and growth

These are infectious to humans and some primates They grow in human cell lines like WI-38 and MRC-5 lines

Antigenic properties

More than 100 serotypes are known

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Pathogenesis and pathology

Virus enters via upper respiratory tract High titers of virus in nasal secretions are found after 2-4 days of infection

After that virus titer falls but illness persists

Replication is limited to surface epithelium of nasal mucosa

It may lead to edema and mild cellular infiltration Nasal secretion increases in quantity and in protein concentration It may occasionally infect lower respiratory tract
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Clinical features

Acute illness last for 7 days A non productive cough may persist for 2-3 weeks Usual symptoms in adults are sneezing, nasal obstruction, nasal discharge, Sore throat, mild cough, head ache, malaise and chilly sensation, little or no fever Nasal and nasopharyngeal mucosa become red and swollen

Secondary bacterial infections may occur leading to otitis media, sinusitis, bronchitis or pneumonitis in children.
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Epidemiology

It occur throughout the world

Prevalence is lower in summer Virus is transmitted by close contact

The fingers of a person with a cold and transmitted to others

By hand to hand Hand to object to hand contamination.

Self inoculation after hand contamination is the main mode of spread Infection rate is higher in infants, children
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Treatment & control

No specific prevention method or treatment is available

It is difficult to develop a vaccine because of multiplicity of serotypes of rhinoviruses

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CORONAVIRUSES

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Properties of Coronaviruses

Large 80 to 220nm, enveloped RNA viruses

spherical,

There are 20nm long petal shaped projections (spikes) on the outer surface of the envelope like a solar corona The spikes are made up of glycoproteins

They cause common gastroenteritis in infants.

They show recombination high

colds,

These viruses are difficult to culture. frequency of

SARS Severe acute respiratory syndrome is associated with this virus

Replication: takes place in cytoplasm of cell

Genome consists of the longest known, 35

Pathogenesis

Coronaviruses infect humans and other animals

In humans, the infections limited to upper respiratory tract restricted to the ciliated epithelia of the nose and trachea
Responsible for about 30% of common cold infections The infection may lead to patchy destruction of ciliated epithelial cells and the loss of beating cilia It may cause gastroenteritis
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Pathogenesis contd

The SARS virus is transmitted aerogenically

An incubation time of two to 10 days

Clinically, fever and a marked shortness of breath is noted

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Clinical features:

Respiratory disease:

They cause common colds with afebrile, in adults The symptoms are similar to rhinoviruses typically with nasal discharge and malaise Incubation period is 2 to 5 days Lower respiratory infection with pneumonia is also common SARS is a recent syndrome caused by this virus which is characterized by atypical pneumonia

Gastrointestinal Disease:

Coronavirus like particles have been observed in feces of patients with enteritis
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Laboratory diagnosis:

Isolation and identification of virus:

It is very difficult to culture. The antigens in cells in respiratory secretions may be detected by ELISA. Electron microscopy (EM) and PCR can be used to detect coronavirus particles in feces Serodiagnosis using acute and convalescent sera is the main method of confirmation of coronavirus infections ELISA and haemagglutination tests can be used

Direct examination:

Serology:

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Epidemiology

They are major cause of respiratory illness in adults during winter months

When rhinovirus and other respiratory virus infections are low

They have found world wide.

SARS patients should be quarantined.

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Treatment and prevention

The only preventive measure to date is exposure prevention

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REOVIRIDAE

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REOVIRIDAE

The name reovirus is derived from the abbreviation for respiratory enteric orphan virus

No diseases were associated with the virus upon its discovery (hence orphan virus)

This family consists of

Reoviruses: they are not known to cause any human disease Rotavirus Coltiviruses causes Colorado tick fever

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General characteristics

Reoviruses and Rotavirus are 60-80nm with double stranded segmented RNA genome
Capsid is composed of two layered shells The capsid is like wheel (rota means wheel) The virion is icosahedral in symmetry without envelope The family includes human rotaviruses, the most important cause of infantile gastroenteritis around the world

Replication occurs in cytoplasm of host cell

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Rotaviruses

It causes infantile diarrhea and sometimes in adults

Ranks sixth among the worlds global killers

Antigenic properties of rotaviruses:

There are three major subgroups and nine serotypes exists.

Outer capsid protein are important antigens

Rotaviruses associated with human gastroenteritis are classified as group A rotaviruses. But antigenically distinct rotaviruses have also caused diarrheal outbreaks in adults They are fastidious agents to culture
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Cell culture:

Pathogenesis:

Rotaviruses infect cells in villi of small intestine

They multiply in cytoplasm of enterocytes and damage their transport mechanisms

A Rotavirus encoded protein may act as viral enterotoxin and induces secretion by triggering signal transduction pathway Damaged cells slough into lumen of intestine and release large quantities of virus in the stool Viral excretion may last up to 2 12 days It may be prolonged in malnourished patients Diarrhea is absorption due to impaired sodium and glucose
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Clinical features

Incubation period is 1 to 4 days

Typical symptoms include diarrhea, fever, abdominal pain, vomiting leading to dehydration In infants and children, severe loss of electrolytes and fluids may be fatal unless treated Asymptomatic infections may occur Common source of infection is contact with pediatric cases

Epidemics in adults have occurred.

Group B rotaviruses are associated with some outbreaks of diarrhea in adult patients in some countries.
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Laboratory diagnosis

Demonstration of virus in stool collected in early in illness and on a rise in antibody titer

Virus can be observed by IEM (immune electron microscopy), immunodiffusion, ELISA or PCR

Antibody titer rise can be detected by ELISA.

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Epidemiology

Rotavirus infections predominate during winter seasons

80% of the population have antibody against rotavirus by the age of 3

Symptomatic infections are most common in children between 6 months and 2 years
Transmission is by fecal-oral route Nosocomial infections are frequent.
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Annual Rotavirus diarrhea deaths in the world ( Much greater in developing countries)

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Treatment & Control:

It is supportive, to correct the loss of water and electrolytes that may lead to dehydration, acidosis, shock and death Management consists of replacement restoration of electrolyte balance

of

fluids

and

Either intravenously or orally

Live attenuated vaccines now available for use in children

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Norwalk virus

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Norwalk Virus

Norwalk genus name for original Norwalk virus and other Norwalk-like viruses. Family Calicivirus Calicivirae found worldwide, infecting humans, primates, and cattle, among others.

Increasingly being recognized as leading cause of food borne illness.

Occur frequently at institutional settings, Schools, nursing homes. Restaurants, dorms. Crowded quarters and poor hygiene contribute to its spread.
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Properties

Nonenveloped capsid/nucleocapsid.

Icosahedral, 35-39 nm diameter and positive sense RNA Virus has four serotypes
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Mode of transmission

The virus found in stool and vomit of infected.

Very infectious few virus particles needed to start large outbreak. Primarily fecal-oral. Vomit airborne particles,capable of covering large radius

Very contagious infection via eating contaminated food

Contact with sick individual or contaminated surfaces

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Clinical features

Acute gastroenteritis.

Illness begins suddenly, from 12-48 hours after ingestion. Brief illness period.
Very young, elderly, and those with weakened immune systems may experience more severe symptoms. Infectiousness may last up to 2 weeks, no evidence of long-term carriers

Antibody developed has a protective role for short term against reinfection with the same virus
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Diagnosis

Electron Microscope
RT-PCR ELISA Calicivirus is not easily studied

No efficient techniques have been developed to culture it in a lab setting.

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Treatment and Prevention

Treatment is symptomatic
Effective hand washing, careful processing of food is important to prevent food borne outbreaks Purification of drinking water decreases outbreaks

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Astroviruses

Small RNA viruses, ss +, none- enveloped, named because of star-shaped surface morphology, 28 nm in diameter. Associated with cases of endemic gastroenteritis, usually in young children and neonates Can cause occasional outbreaks. Responsible for up to 10% of cases of gastroenteritis. Similar disease to rota and adenoviruses.

Most people have antibodies by the age of three

Diagnosed by electron microscopy only, often very difficult because of small size.
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