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Viral Gastroenteritis

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Viral Gastroenteritis viral gastroenteritis What causes viral gastroenteritis? y Rotaviruses y Caliciviruses y Astroviruses y SRV (Small Round Viruses) y Toroviruses y Adenoviruses 40 , 41 Diarrhea Causing Agents in World ROTAVIRUS Family Reoviridae Genus Segments Host Vector Orthoreovirus 10 Mammals None Orbivirus 11 Mammals Mosquitoes, flies Rotavirus 11 Mammals None Coltivirus 12 Mammals Ticks Seadornavirus 12 Mammals Ticks Aquareovirus 11 Fish None Idnoreovirus 10 Mammals None Cypovirus 10 Insect None Fijivirus 10 Plant Planthopper Phytoreovirus 12 Plant Leafhopper OiOryzavirus 10 Plan t Plan thopper Mycoreovirus 11 or 12 Fungi None? REOVIRUS y REO: respiratory enteric orphan, y early recognition that the viruses caused respiratory and enteric infections y incorrect belief they were not associated with disease, hence they were considered "orphan " viruses ROTAVIRUS‐ PROPERTIES y Virus is stable in the environment (months) y Relatively resistant to hand washing agents y Susceptible to disinfection with 95% ethanol, ‘Lyy,sol’, formalin STRUCTURAL FEATURES OF ROTAVIRUS y 60‐80nm in size y Non‐enveloped virus y EM appearance of a wheel with radiating spokes y Icosahedral symmetry y Double capsid y Double stranded (ds) RNA in 11 segments Rotavirus structure y The rotavirus genome consists of 11 segments of double- stranded RNA, which code for 6 structural viral proteins, VP1, VP2, VP3, VP4, VP6 and VP7 and 6 non-structural proteins, NSP1-NSP6 , where gene segment 11 encodes both NSP5 and 6. y Genome is encompassed by an inner core consisting of VP2, VP1 and VP3 proteins. Intermediate layer or inner capsid is made of VP6 determining group and subgroup specifici ti es. y The outer capsid layer is composed of two proteins, VP7 and VP4 eliciting neutralizing antibody responses. CLASSIFICATION‐ Groups y 7 Groups (A through G) and Within group A, four different subgroups (SG); SGI, SGII, SGI and II, and nonI/nonII, have been dis tingui sh ed on th e b asi s of VP6 divers ity, among whic h poss ibly SGI or SGII are the only humans strains y Group A is the most common y Groupp( B (outbreaks in China) y Group C (worldwide) CLASSIFICATION ‐‐ Serotypes y Serotypes based on viral capsid proteins inducing neutralizing Ab y 15 G serotypes based on VP7 (G serotypes) differences y 5 predominant strains in U.S. (G1-G4, G9) account for 90% of isolates y Strain G1 accounts for 73% of infections y 25 P serotypes based on VP4 (P serotypes) with P4/P8 predominance y Common PG combinations are: P8G1, P8G2, P4G2, P8G4 Rotavirus gene function Rotavirus Pathogenesis y Entry through mouth y Targeted host cells - mature enterocytes lining the tips of intestinal villi y Intermediate/infective sub-viral particle (ISVP) produced throug h pro teol ysi s y Enter host cell by endocytosis y Replication in epithelium of small intestine y Replication outside intestine and viremia uncommon y Infection leads to isotonic diarrhea CLINICAL FEATURES y Incubation period ‐ thought to be <4 days y Fever‐ can be higggh grade ( >102°F in 30%) y Vomiting, nausea precede diarrhea y Diarr hea MECHANISM OF DIARRHEA Watery diarrhea due to net secretion of intestinal fluid and loss of absorptive surface y Activation of the enteric nervous system y Role of NSP4 peptide regions as an enterotoxin y Dehydration is the main contributor to mortality y Secondary malabsorption of lactose and fat , and chronic diarrhea are possible EPIDEMIOLOGY y Age‐ children 4mo - 2 years y Asymptomatic infections are common, especially in adults y Nosocomial infections y Outbreaks y Seve re Disease yyg,oung, immunocom promised RRtotavi rus IIitmmunity y Antibody against VP7 and VP4 partially protective y First infection usually does not lead to permanent immunity y Reinfection can occur at any age y Subsequent infections generally less severe Rotavirus detection and strain characterization y Laboratory procedures for diagnosis of rotavirus include : y electron microscopy (EM) , y passive latex agglutination assays (LA), y electropherotyping using polyacylamide gel electropp(),horesis (PAGE), y enzyme-linked immunosorbent assays (ELISA) y reverse transcription-polymerase chain reaction (RT-PCR) TREATMENT AND PREVENTION y Treatment Supportive‐ oral, IV rehydration y Prevention Hand hygiene and disinfection of surfaces y Vaccine Calicivirus HUMAN CALICIVIRUSES ((HuCVHuCV)) • Family Caliciviridae • Non-enveloped RNA viruses with ss [+] sense RNA • 27-35 nm in size • Contains singgpple capsid protein CLASSIFICATION Caliciviruses can be divided into: Norwalk and "Norwalk-like" viruses (NLV) "Sapporo-like" viruses (SLV) Vesiviruses Lagoviruses CLASSIFICATION NLV (Norovirus) SLV (Sapovir us) Norwalk virus Sapporo virus Hawaii virus Manchester virus Snow Mountain virus Houston/86 Montgomery county virus London/92 Taunton (England) Norwalk virus: Clinical Features • 24 hour incubation period • Vomiting prominent • Headache, myalgia, fever • Diarrhea 1-3 days, less severe than rotavirus • Affects all ages • Treatment symptomatic • rehydration, antidiarrheals • Complications rare • immunocompromised SPREAD y Person-to-person fecal-oral spp(read (stool/ vomitus ) y Fecal contamination of food or water y Fit(tl/it)Fomites (stool/ vomitus) EPIDEMIOLOGY y Worldwide distribution y >23 million cases/year in the U.S. ا(y Major cause of food -borne outbreaks of (>50% y Most people have had infections by age 4 years (by sero-prevaldi)lence studies) DIAGNOSIS y diagnosed by electron microscopy only, often difficult to diagnose because of small size y Specimen: stool, vomitus, food, environmental swabs (during outbreak investigations) y RT-PCR iblihlhlbin state public health labs. y Serology for epidemiologic purposes Astrovirus Particles Classification of Astrovirus y Genus Astrovirus y Family Astroviridae y Human serotypes: HuAstV 1-8 ASTROVIRUS‐ structure y Small ss RNA virus y Non-enveloped y 27-32nm in size y Contain 3 structural proteins y Genome has been sequenced ASTROVIRUS ‐ Clini cal Features y Infants and children are most often affected y Elder ly and i mmunecompromi sed persons al so y Short incubation period 1-4 days y Nausea, vomiting, abdominal cramping and watery diarrh ea y Constitutional syypmptoms-fever, malaise, headache ASTROVIRUS ‐ Epidemi ol ogy y Endemic worldwide y Mainly in children < 7 years of age y Transmission- person-to-person [fecal-oral] y Outbreaks due to fecal contamination of sea-food or water ASTROVIRUS ‐ Diagnosis y EM (virus shed in stool in great numbers) y EIA y RT-PCR Toroviruses y Family Coronaviridae y Genus Torovirus y Human and animal pathogen y Pleomorp hic, coa te d ss (+ ) RNA v irus y Watery diarrhea in 2 – 12 months old y Diagnosis: EM Adenovirus Particle Enteric Adenoviruses y Naked DNA viruses, 75 nm in diameter. y fftidiastidious ent tieric ad enovi rus types 40 and 41 are associat ed with gastroenteritis y associated with cases of endemic gastroenteritis, usually in young children and neonates. Can cause occasional outbreaks. y possibly the second most common viral cause of gastroenteritis (7-15% of all endemic cases) y similar disease to rotaviruses y most people have antibodies against enteric adenoviruses by the age of three y diagnosed by EM or by the detection of adenovirus antigens in faeces by ELISA or other assays. Diagnosis of viral diseases Gastrointestinal Viral Infections ¾ Culture: rotaviruses, enteric adenoviruses (especially, serotypes 40 and 41), noroviruses, sapoviruses, astroviruses, and possibly toroviruses that is not readily cultivable. ¾ Electron Microscopy: Negative staining with uranyl acetate or phosphotungstic acid can be performed directly on stool specimens. especially the noroviruses and sapoviruses, are better visualized using immune electron microscopy Gastrointestinal Viral Infections ¾ Antigen Detection: Many commercial assays that use for rotavirus detection, and at least one commercial adenovirus assay is available. Antigen detection assays for astrovirus have been used in investigations ¾ Nucleic Acid Amplification: In rotaviruses antigen detection assays better than RT-PCR because of the adequate sensitivity and greater simplicity . RT-PCR has been shown to be more sensitive than EIA for detection of astroviruses. RT-PCR is the method of choice for detection of noroviruses. Types of Cell C ult ure ¾ Primary cell cultures; ddierived directl y f rom th e source ani mal . incl ud e pri mary monk ey kid ney cell s and primary rabbit kidney cells. ¾ Diploid or semicontinuous cells; are capable of a limited number of passages before undergoing senescence. Include human fibroblast cell cultures such as MRC-5 and WI-38 cells. ¾ Continuous or transformed cell lines; are immortalized cells that can be passaged without limit. include HEp-2, HeLa, A549, and Madin-Darby canine kidney cells. Electron Microscop y ¾for the direct visualization of viral particles in specimen. ¾ Advantages include speed, lack of requirement for viral viability, and that many different kinds of viral particles can potentially be seen (nature of the particle). ¾Disadvantages include the cost and complexity of maitiiintaining an el ect ron mi croscopy, th e need df for a skilled operator, and limited sensitivity related to the relatively high concentration of viral particles . 1. Direct t S peci men E xami nat tiion (EM) ¾diaggygnostic virology is the evaluation of stool specimens from patients with suspected viral gastroenteritis. ¾ rotaviruses, enteric strains of adenovirus, noroviruses, and astroviruses, are not readily cultivable. ¾All be seen by negatively stained using
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