Metabolic Brain Disease

, Volume 33, Issue 4, pp 1165–1173 | Cite as

A novel compound-primed multiplex ARMS-PCR (CPMAP) for simultaneous detection of common PAH gene mutations

  • Maryam Shaykholeslam Esfahani
  • Ehsan Shaykholeslam Esfahani
  • Sadeq Vallian
Original Article


In this study, we introduce a novel compound-primed multiplex ARMS PCR (CPMAP) for simultaneous detection of common PAH gene mutations. This approach was used successfully for simultaneous identification of six most common PAH gene mutations in 137 phenylketonuria patients in the Iranian population. A total of six normal and six mutant allele-specific primers and 4 common primers containing a tag sequence of 12 base pair at the 5ˊ-end were designed and used in two separate optimized multiplex ARMS reactions followed by hot-start PCR. The products were separated and visualized on 3% agarose gel. The CPMAP genotyping data were completely in accordance with the direct sequencing results. The CPMAP suggests a reliable, economical and rapid method for simultaneous detection of PAH point mutations using conventional PCR, which could be applied for diagnosis of other gene mutations.


Multiplex ARMS-PCR Compound primer Phenylalanine hydroxylase mutations Phenylketonuria 



This study was supported by University of Isfahan institutional review board (IRB) for research and ethics approval (Approval Ref No. 790205). We are immensely grateful to Dr. Austin J Carlson for constructive and thorough proofreading and English language editing of our manuscript.


This study was supported by University of Isfahan institutional review board (IRB) for research and ethics approval (Approval Ref No. 790205).

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Maryam Shaykholeslam Esfahani
    • 1
  • Ehsan Shaykholeslam Esfahani
    • 2
  • Sadeq Vallian
    • 1
  1. 1.Division of Genetics, Department of Biology, Faculty of ScienceUniversity of IsfahanIsfahanIran
  2. 2.Division of Biotechnology, Department of Agricultural Biotechnology, Faculty of AgricultureIsfahan University of TechnologyIsfahanIran

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