Scribd Logo
Upload

Welcome to Scribd!

  • 25 views

    Viral Cultures: Investigation Strategies and Methods

    Uploaded by

    Diyantoro Nyo
    This document discusses viral culture techniques for identifying viruses. It describes how viral cultures work by amplifying viruses through intracellular replication. The document outlines various techniques for virus identification including PCR, electron microscopy, and antigen capture. It also details the virus culture process, including obtaining suitable specimens, transporting them while maintaining a cold chain, selecting appropriate cell cultures, and detecting viruses through cytopathic effects, immunological methods, or sequencing. The document notes that virus culture is a long process and not always suitable for front-line diagnosis.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd

    Viral Cultures: Investigation Strategies and Methods

    Uploaded by

    Diyantoro Nyo
    25 views12 pages
    This document discusses viral culture techniques for identifying viruses. It describes how viral cultures work by amplifying viruses through intracellular replication. The document outlines various techniques for virus identification including PCR, electron microscopy, and antigen capture. It also details the virus culture process, including obtaining suitable specimens, transporting them while maintaining a cold chain, selecting appropriate cell cultures, and detecting viruses through cytopathic effects, immunological methods, or sequencing. The document notes that virus culture is a long process and not always suitable for front-line diagnosis.

    Original Title

    ihr_l10virusculture_en.ppt

    Copyright

    © © All Rights Reserved

    Available Formats

    PPT, PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    This document discusses viral culture techniques for identifying viruses. It describes how viral cultures work by amplifying viruses through intracellular replication. The document outlines various techniques for virus identification including PCR, electron microscopy, and antigen capture. It also details the virus culture process, including obtaining suitable specimens, transporting them while maintaining a cold chain, selecting appropriate cell cultures, and detecting viruses through cytopathic effects, immunological methods, or sequencing. The document notes that virus culture is a long process and not always suitable for front-line diagnosis.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    25 views12 pages

    Viral Cultures: Investigation Strategies and Methods

    Uploaded by

    Diyantoro Nyo
    This document discusses viral culture techniques for identifying viruses. It describes how viral cultures work by amplifying viruses through intracellular replication. The document outlines various techniques for virus identification including PCR, electron microscopy, and antigen capture. It also details the virus culture process, including obtaining suitable specimens, transporting them while maintaining a cold chain, selecting appropriate cell cultures, and detecting viruses through cytopathic effects, immunological methods, or sequencing. The document notes that virus culture is a long process and not always suitable for front-line diagnosis.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    You are on page 1of 12

    Investigation strategies and methods

    Viral cultures

    May 2007

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    Learning objectives

    At the end of the presentation, participants should:


    • Understand the principle of cultivating viruses

    • Understand the methods and problems with cultivating viruses

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    Techniques to identify viruses
    It can take a few hours to weeks to identify a virus

    Techniques include:
    • PCR (single round) or nested/semi-nested PCR
    • Real-time PCR
    • Direct electronic microscopy
    • Antigen capture
    • Isolation
    – Long process

    – Gold standard for viruses that can be cultured


    Laboratory Training for FieldEEpidemiologists
    P I D E M I C A L E R T A N D R E S P O N S E
    Virus culture
    Is based upon amplification of potentially infectious pathogens

    Implies intracellular replication of viruses in the cytoplasm or


    in the nucleus

    Is controlled by regulations (i.e. bio-safety level 2, 3 or 4)

    Allows for:

    • Identification

    • Further studies
    (e.g., Pathogenicity, antiviral sensitivity, research)

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    Virus culture

    Long process
    • Not always possible for front-line diagnosis

    Primary objective for the diagnosis of an unknown disease

    No generic protocol

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    How to go about virus culture?
    Obtain suitable specimens

    • Identified specimens with suitable information

    Evaluate of chances of success of the process before start

    Make sure transportation used cold chain

    • 4°C

    • -20°C

    • Dry ice (-79°C)

    Use suitable culture protocol

    • In vitro/in vivo cell cultures

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    Culture procedure
    Use of a variety of cell sources and techniques

    Treatment of the specimen prior to inoculation

    Follow-up

    Viral detection:

    • Non specific

    – Cytopathogenic effect (microscope)

    – Electronic microscopy identification (morphology)

    • Specific

    – Immunological detection: antigen detection, PCR, IFA…

    Viral load estimation (titration, plaque assay)


    Laboratory Training for FieldEEpidemiologists
    P I D E M I C A L E R T A N D R E S P O N S E
    Limitations of cultures to
    identify viruses
    Absence of detection system for the agent

    Inappropriate culture systems

    Viruses that cannot be cultured

    A negative viral culture results does not mean that the


    agent is absent
    • Need of other tests

    • PCR can detect the viral genome in absence of the


    complete virus

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    Specimens used to culture viruses

    Blood specimens Stools, rectal swabs

    • EDTA Urine
    • Heparin Saliva
    • Serum
    Cerebro-spinal fluid
    Stool
    Biopsy
    Throat swabs • Skin (filoviridae)
    Naso-paryngeal aspirates • Organs (fixation with
    formaldehyde 10%)

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    Potentially infectious specimen forms

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    Sequencing
    Analysis of sequence of nucleic acid fragment after
    PCR amplification

    Comparison of the alignment of nucleotides with other


    sequences present in different data bases for the
    identification of an agent

    Confirmatory analysis
    • Final DNA fingerprint is molecular signature of the micro-organism

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E
    Investigation strategies and methods

    Developed by the Department of Epidemic and


    Pandemic Alert and Response of the World Health
    Organization with assistance from:

    European Program for Intervention


    Epidemiology Training

    Canadian Field Epidemiology Program

    Thailand Ministry of Health

    Institut Pasteur

    Laboratory Training for FieldEEpidemiologists


    P I D E M I C A L E R T A N D R E S P O N S E

    You might also like

    pFad - Phonifier reborn

    Pfad - The Proxy pFad of © 2024 Garber Painting. All rights reserved.

    Note: This service is not intended for secure transactions such as banking, social media, email, or purchasing. Use at your own risk. We assume no liability whatsoever for broken pages.


    Alternative Proxies:

    Alternative Proxy

    pFad Proxy

    pFad v3 Proxy

    pFad v4 Proxy