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By:
Denisse Class
      &
Nirzari Amin
   Rotavirus is a non-enveloped virus that comes
    from a family called Reoviridai.
   Derives the name from the Latin Rota which
    means wheel. It appears wheel like when looked
    under an electric microscope.
   Very stable and may remain viable for weeks or
    months if not disinfected
   Once cured, it can reoccur at any age. But the
    subsequent infections are usually less severe
   Can cause:
       Gastroenteritis
       Dehydration
   This virus can easily be spread through children
    before and/or after becoming sick with diarrhea.
   Usually transmitted by fecal-oral route
       Virus must be shed of an infected person and somehow
        entered into a susceptible person to cause infection.
   Also caused by contaminated hands, water, and,
    objects.
   It has been observed that improved sanitation
    does not decrease the infection transmission.
   Due to this, the primary treatment is vaccination.
       Few current examples include; RotaTeq, Rotatrix
   No antiviral drug has yet been discovered.
   Treatment is nonspecific but usually treated for
    dehydration.
   Symptoms usually appear within 12 to 48 hrs.
    after exposure and last till 1-3 days.
     Watery diarrhea
     Vomiting
     Fever
     Headache
     Abdominal pain
     Chills
   Identification of etiologic agent
   Based on symptoms and physical exam
   Elisa test
   Latex agglutination (for rapid results)
   Elisa
   PCR
   This is an in-vitro test for the       Limitations:
    determination of rotavirus               Stool sample varies by patient
    antigen in feces.                        Patients with infection for a long
                                              period of time results will not
   Polyclonal anti-rotavirus and             show as an active infection.
    anti-rotavirus monoclonal                Early treatment with proper
    antibodies are used.                      antibiotics.
   Specificity – 97%
   Sensitivity – 100%
   Expectations:
     Healthy individuals should test
      negative.
     Presence of rotavirus antigen
      should be present .
   Break the wells need it for
    samples plus two controls.
   Prepare controls.
   Add 100 µL of specimen,
    incubate for 30 min.
   Add 2 drops of reagent 1 Blue
    sol. to each well, incubate for 5
    min. then wash
   Add 2 drops of reagent 2 red
    sol. incubate for 5 min and then
    wash.
   Add 2 drops of chromogen sol.
    @room temp. for 5 min. DO
    NOT WASH.
   Add 2 drops of stop sol. Mix
    then read results .
   Limitations:                      Expected results:
     For the making of the             Amplification/Extension
      new template the                   of DNA.
      primers have to be very           Allows scientists to
      specific and particular to         sequence nucleotides.
      the template.                      These nucleotides are
     The primers must be                very helpful in studying
      only capable of binding            the molecular
      with the template and              epidemiology of
      not each other because             Rotavirus.
      this can cause the
      amplification of a
      short, useless DNA.
ELISA                      PCR

   97% specificity             Less specific
   100% sensitivity            Super sensitive
   Fluid sample                Fluid sample
   Results in 45-60 min        2 to 3 hours
   www.rapidtest.com/pdf/Rotavirus_8306-3.pdf
   http://www.hopkinsmedicine.org/heic/ID/rotavirus/
   http://www.microbac.com/services/pcr_process.php

   Brasier,Alan R., Adolfo Garcia-Sastre, A, Lemon, S. M., & ebrary, I.
    (2009).
       Cellular signaling and innate immune responses to RNA virus
       infections. Washington, D.C. : ASM Press.
   Wark, P. (2008). Paediatric Respiratory Reviews. Mini-symposium:
    Innate immunity in the lung. Paediatric Respiratory Reviews, 9(4),
    233-235. Retrieved from EBSCOhost.
   Rotavirus. In http://www.cdc.gov. Retrieved 09/26/2011, from
    http://www.cdc.gov/rotavirus/index.html.
   Scott M, Janneck L, Dent R, Merino D. Rotavirus. Retrieved from
    http://www.brown.edu/Courses/Bio_160/Projects2004/rotavirus/
    Pathology.htm
   http://faculty.plattsburgh.edu/donald.slish/PCR.html
   http://www.humenhealth.com/rotavirus-infection/rotavirus-
    infection.asp
   http://www.humenhealth.com/rotavirus-2

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Rotavirus Detection & Diagnosis

  • 1. By: Denisse Class & Nirzari Amin
  • 2. Rotavirus is a non-enveloped virus that comes from a family called Reoviridai.  Derives the name from the Latin Rota which means wheel. It appears wheel like when looked under an electric microscope.  Very stable and may remain viable for weeks or months if not disinfected  Once cured, it can reoccur at any age. But the subsequent infections are usually less severe
  • 3. Can cause:  Gastroenteritis  Dehydration
  • 4. This virus can easily be spread through children before and/or after becoming sick with diarrhea.  Usually transmitted by fecal-oral route  Virus must be shed of an infected person and somehow entered into a susceptible person to cause infection.  Also caused by contaminated hands, water, and, objects.
  • 5. It has been observed that improved sanitation does not decrease the infection transmission.  Due to this, the primary treatment is vaccination.  Few current examples include; RotaTeq, Rotatrix  No antiviral drug has yet been discovered.  Treatment is nonspecific but usually treated for dehydration.
  • 6. Symptoms usually appear within 12 to 48 hrs. after exposure and last till 1-3 days.  Watery diarrhea  Vomiting  Fever  Headache  Abdominal pain  Chills
  • 7.
  • 8.
  • 9. Identification of etiologic agent  Based on symptoms and physical exam  Elisa test  Latex agglutination (for rapid results)
  • 10. Elisa  PCR
  • 11. This is an in-vitro test for the  Limitations: determination of rotavirus  Stool sample varies by patient antigen in feces.  Patients with infection for a long period of time results will not  Polyclonal anti-rotavirus and show as an active infection. anti-rotavirus monoclonal  Early treatment with proper antibodies are used. antibiotics.  Specificity – 97%  Sensitivity – 100%  Expectations:  Healthy individuals should test negative.  Presence of rotavirus antigen should be present .
  • 12. Break the wells need it for samples plus two controls.  Prepare controls.  Add 100 µL of specimen, incubate for 30 min.  Add 2 drops of reagent 1 Blue sol. to each well, incubate for 5 min. then wash  Add 2 drops of reagent 2 red sol. incubate for 5 min and then wash.  Add 2 drops of chromogen sol. @room temp. for 5 min. DO NOT WASH.  Add 2 drops of stop sol. Mix then read results .
  • 13. Limitations:  Expected results:  For the making of the  Amplification/Extension new template the of DNA. primers have to be very  Allows scientists to specific and particular to sequence nucleotides. the template. These nucleotides are  The primers must be very helpful in studying only capable of binding the molecular with the template and epidemiology of not each other because Rotavirus. this can cause the amplification of a short, useless DNA.
  • 14.
  • 15. ELISA PCR  97% specificity  Less specific  100% sensitivity  Super sensitive  Fluid sample  Fluid sample  Results in 45-60 min  2 to 3 hours
  • 16. www.rapidtest.com/pdf/Rotavirus_8306-3.pdf  http://www.hopkinsmedicine.org/heic/ID/rotavirus/  http://www.microbac.com/services/pcr_process.php  Brasier,Alan R., Adolfo Garcia-Sastre, A, Lemon, S. M., & ebrary, I. (2009). Cellular signaling and innate immune responses to RNA virus infections. Washington, D.C. : ASM Press.  Wark, P. (2008). Paediatric Respiratory Reviews. Mini-symposium: Innate immunity in the lung. Paediatric Respiratory Reviews, 9(4), 233-235. Retrieved from EBSCOhost.
  • 17. Rotavirus. In http://www.cdc.gov. Retrieved 09/26/2011, from http://www.cdc.gov/rotavirus/index.html.  Scott M, Janneck L, Dent R, Merino D. Rotavirus. Retrieved from http://www.brown.edu/Courses/Bio_160/Projects2004/rotavirus/ Pathology.htm  http://faculty.plattsburgh.edu/donald.slish/PCR.html  http://www.humenhealth.com/rotavirus-infection/rotavirus- infection.asp  http://www.humenhealth.com/rotavirus-2