Evaluation of multiplex SYBR green real-time PCR assay for detection of pathogenic Escherichia coli

  • Authors

    • Ema Komalasari University of Al Azhar Indonesia
    • Winiati P. Rahayu IPB University
    • Siti Nurjanah IPB University
    2020-04-28
    https://doi.org/10.14419/ijet.v9i2.30389
  • Melting Curve, Multiplex Rt-PCR, Pathogenic E. coli, Specificity, SYBR Green.

  • Abstract

    Pathogenic Escherichia coli (E. coli) has been implicated in a wide range of disease causing infections. It is essential to generate a method for detecting and differentiating each pathotype of E. coli which is more quickly and efficiently by using less reagent. This study aimed to evaluate a SYBR Green multiplex real-time PCR method for detecting four types of pathogenic E. coli. Two of multiplex real-time PCR system, 6-plex and 3-plex, were set to detect six different virulence factors from ETEC, EPEC, EHEC, and EIEC and evaluate the melting curves and specificity compared to simplex method. The results showed that 3-plex rt-PCR method gave more reliable melting curves than 6-plex. The 3-plex rt-PCR also provided similar melting value (Tm) to simplex system. The results of this specificity assay supported the selection of 3-plex rt-PCR conditions for detection of pathogenic E. coli.

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  • How to Cite

    Komalasari, E., P. Rahayu, W., & Nurjanah, S. (2020). Evaluation of multiplex SYBR green real-time PCR assay for detection of pathogenic Escherichia coli. International Journal of Engineering & Technology, 9(2), 448-453. https://doi.org/10.14419/ijet.v9i2.30389

    Received date: 2020-02-07

    Accepted date: 2020-04-04

    Published date: 2020-04-28