Location via proxy:   [ UP ]  
[Report a bug]   [Manage cookies]                
Scribd Logo
Upload

Welcome to Scribd!

  • 17 views

    Salmonella Enteritidis in Egg and Meat Based On Real-Time PCR

    Uploaded by

    Amir Mahmud
    1) The document describes the development of a rapid detection kit for the food-borne pathogen Salmonella enteritidis in eggs and meat using real-time PCR. 2) The researchers designed primers for the prot6E gene of Salmonella enteritidis and optimized real-time PCR conditions. 3) Testing showed the primers specifically detected Salmonella enteritidis with high sensitivity down to a concentration of 43.2 pg/μL, and could detect the pathogen in artificially contaminated egg and meat samples.

    Copyright:

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

    Salmonella Enteritidis in Egg and Meat Based On Real-Time PCR

    Uploaded by

    Amir Mahmud
    17 views15 pages
    1) The document describes the development of a rapid detection kit for the food-borne pathogen Salmonella enteritidis in eggs and meat using real-time PCR. 2) The researchers designed primers for the prot6E gene of Salmonella enteritidis and optimized real-time PCR conditions. 3) Testing showed the primers specifically detected Salmonella enteritidis with high sensitivity down to a concentration of 43.2 pg/μL, and could detect the pathogen in artificially contaminated egg and meat samples.

    Original Title

    NB 8 Putri Anisa

    Copyright

    © © All Rights Reserved

    Available Formats

    PPTX, PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    1) The document describes the development of a rapid detection kit for the food-borne pathogen Salmonella enteritidis in eggs and meat using real-time PCR. 2) The researchers designed primers for the prot6E gene of Salmonella enteritidis and optimized real-time PCR conditions. 3) Testing showed the primers specifically detected Salmonella enteritidis with high sensitivity down to a concentration of 43.2 pg/μL, and could detect the pathogen in artificially contaminated egg and meat samples.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    17 views15 pages

    Salmonella Enteritidis in Egg and Meat Based On Real-Time PCR

    Uploaded by

    Amir Mahmud
    1) The document describes the development of a rapid detection kit for the food-borne pathogen Salmonella enteritidis in eggs and meat using real-time PCR. 2) The researchers designed primers for the prot6E gene of Salmonella enteritidis and optimized real-time PCR conditions. 3) Testing showed the primers specifically detected Salmonella enteritidis with high sensitivity down to a concentration of 43.2 pg/μL, and could detect the pathogen in artificially contaminated egg and meat samples.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    of 15

    Development of Rapid Kit Detection Food-borne Pathogen

    Salmonella enteritidis in Egg and Meat Based on Real-Time PCR

    Dr. Muktingsih Nurjayadi, M.Si


    Putri Anisa Nila Auni
    Hafiza Muslimah
    Vira Saamia, S.Si, M.BioMed

    Department of Chemistry, Faculty of Mathematics


    and Natural Science, Universitas Negeri Jakarta,
    Indonesia.
    Salmonellosis take over 30%
    out of death case in total as
    an impact of food borne
    pathogen in United States.

    25% out of 100%


    outbreak cases in
    Indonesia is food
    poisoning.
    Real-Time PCR
    Offers a rapid, sensitive, and specific detection for
    microorganisms until low concentration.
    Workflow

    Preparation of Primer Design


    culture
    sample

    Confirmation
    assay of meat and
    egg samples
    DNA Isolation

    Artificial
    Real-Time
    contaminated egg
    PCR
    and meat samples
    Amplification
    with Salmonella
    Assay
    enteritidis
    1 2 3 4 5 6 7 8
    The visualization is using 2%
    1500bp
    Agarose Gel Electrophoresis
    1000bp
    (1) DNA Ladder 100 bp; (2)
    Negative control; (3) Posiive
    control fimC Salmonella 500bp
    typhi 95bp; (4) DNA
    300bp
    fragment at temperature
    200bp 170bp
    570C; (5) DNA fragment at
    temperature 580C; (6) DNA 100bp
    fragment at temperature
    590C; (7) DNA fragment at
    temperature 600C; (8) DNA
    fragment at temperature
    610C.

    Optimization annealing
    prot6E primer pairs
    Confirmation analysis prot6E gene of
    Salmonella enteritidis

    Get a modern PowerPoint Presentation that is


    beautifully designed. I hope and I believe that this
    Template will your Time, Money and Reputation. Easy to
    change colors, photos.

    Melting curve confirmation prot6E-f dan Stock culture Negative


    prot6E-r in Salmonella enteritidis. Melting amplification at control
    temperature that produced is 75,7560C. PCR cycles (NTC)
    13,11 and 13, 69
    Spesification assay prot6E gene of Salmonella
    enteritidis with non-target bacteria

    Tm
    Primer Line Template Ct
    (0C)
    Prot6E
    Salmonella
    Salmonell 23,400 82,98
    a typhimurium
    enteritidis
    Salmonella typhi 21.135 76,34
    Get a modern PowerPoint Presentation that is
    beautifully designed. I hope and I believe that this
    Shigella
    Template will your Time, Money and dysentriae
    Reputation. Easy21.367
    to 76,34
    change colors, photos.
    Klebsiella
    20.350 82,31
    pneumoniae
    Staphylococcus
    17.492 82,36
    aureus
    Vibrio
    19,775 79,67
    parahemolyticus

    Specificity results of prot6E with non-target Specificity amplification curve prot6E gene.
    bacteria
    Sensitivity assay prot6E gene of Salmonella
    enteritidis.

    Line Konsentrasi (pg/µL) Ct

    27000 20,940

    5400 23,746

    1080 26,254

    216 28,570

    43.2 29,931

    Amplification curve of sensitivity prot6E Sensitivity assay results of prot6E primer pairs
    Salmonella enteritidis at various DNA concentration.
    DNA template still can be
    detected at a lower
    concentration of 43.2 pg/µL at
    Ct 29.931 which is equivalent
    to 0.153 CFU/mL

    Standard curve. Shows that the equation is


    y = -2.7306x+27,71. With R2 0,7396.

    Standard curve of prot6E gene


    of Salmonella enteritidis.
    Confirmation analysis prot6E gene in
    meat samples

    Get a modern PowerPoint Presentation that is


    beautifully designed. I hope and I believe that this
    Template will your Time, Money and Reputation. Easy to
    change colors, photos.

    Melting curve meat samples. Both positive


    control and 10-5 contaminated sample were on 10-5 dilution Negative
    sample control (NTC)
    the same melting temperature, 76,9710C.
    Negative control has a different melting Positive control
    temperature, 78,8710C. (contaminated)
    Confirmation analysis prot6E
    gene in egg sample

    Get a modern PowerPoint Presentation that is


    beautifully designed. I hope and I believe that this
    Template will your Time, Money and Reputation. Easy to
    change colors, photos.

    Melting Curve In egg Samples. Both positive Positive


    control and 10-5 contaminated sample were on Negative
    control at Ct control (NTC)
    the same melting temperature, 76,9710C. 10,91
    Negative control has a different melting
    10-5 dilution sample
    temperature, 78,8710C.
    Acknowledgement

    This research is funded by Ministry of Research,


    Technology, and Higher Educaton (Indonesia by
    the scheme PDUPT under the agreement
    contract no. 2/SP2H/DRPM/LPPM-
    UNJ/III/2019.
    Conclusion
    Real-Time PCR assay show that prot6E primer pairs has been
    successes to detect Salmonella enteritidis in both culture stock, and
    artificial contaminated food samples. Based on this result, it can be
    develop as a rapid kit detection for Salmonella enteritidis. Specificity
    assay conclude that prot6E-f and prot6E-r are specifically for target
    bacteria, Salmonella enteritidis. And these primer pairs could amplify
    bacterial DNA at lowest concentration 43. 2pg/µL. This method could
    be develop as a detection kit model to detect pathogenic bacteria as a
    rapid, sensitive, and specific and show an accurate result.
    Our research team
    Thank you
    You can find me at :
    putriauni10@gmail.com

    You might also like