Ann Lab Med.  2021 Jul;41(4):394-400. 10.3343/alm.2021.41.4.394.

Accuracy and Performance Evaluation of Triplet Repeat Primed PCR as an Alternative to Conventional Diagnostic Methods for Fragile X Syndrome

  • 1Department of Laboratory Medicine, Graduate School, Kyung Hee University, Seoul, Korea
  • 2Department of Laboratory Medicine, Seoul National University Hospital, Seoul National University College of Medicine, Seoul, Korea
  • 3Department of Laboratory Medicine, Kosin University Gospel Hospital, Busan, Korea
  • 4Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea


Conventional diagnosis of fragile X syndrome (FXS) is based on a combination of fragment analysis (FA) and Southern blotting (SB); however, this diagnostic approach is time- and labor-intensive and has pitfalls such as the possibility of missing large number alleles. Triplet repeat primed PCR (TP-PCR) is a current alternative used to overcome these limitations. We evaluated the diagnostic usefulness of TP-PCR compared with the conventional diagnostic protocol consisting of FA and/or SB in terms of allele categorization, repeat number correlation, and zygosity concordance in female genetic carriers.
From November 2013 to March 2018, 458 patients (326 males, 132 females) were simultaneously examined using FA and/or SB and TP-PCR by detecting CGG repeat numbers in FMR gene and diagnosed as per American College of Medical Genetics guidelines.
The TP-PCR results showed high concordance with the FA and/or SB results for all three aspects (allele categorization, repeat number correlation, and zygosity concordance in female genetic carriers). TP-PCR detected CGG expansions ≥ 200 in all full mutation (FM) allele cases in male patients, as well as both the normal allele (NL) and FM allele in female carriers. In premutation (PM) allele carriers, the TP-PCR results were consistent with the FA and/or SB results. In terms of zygosity concordance in female genetic carriers, 12 NL cases detected by TP-PCR showed a merged peak consisting of two close heterozygous peaks; however, this issue was resolved using a 10-fold dilution.
TP-PCR may serve as a reliable alternative method for FXS diagnosis.


Fragile X syndrome; Fragment analysis; Southern blotting; Triplet repeat primed PCR; CGG expansion; Allele categorization; Repeat number; Zygosity
Full Text Links
  • ALM
export Copy
  • Twitter
  • Facebook
Similar articles
    DB Error: unknown error