Method-Based Proficiency Test Program for Assessing Quality of Sanger Sequencing-Based Molecular Tests
10.15263/jlmqa.2025.47.1.28
- Author:
Moon-Woo SEONG
1
;
Ho-bin SUNG
;
Jee-Soo LEE
Author Information
1. Cancer Research Institute, Seoul National University, Seoul, Korea
- Publication Type:ORIGINAL ARTICLE
- From:Journal of Laboratory Medicine and Quality Assurance
2025;47(1):28-31
- CountryRepublic of Korea
- Language:English
-
Abstract:
Background:Sanger sequencing is a technology used to identify the gene sequence variants causing rare genetic disorders. However, designing and implementing a proficiency scheme for Sanger sequencing-based genetic testing is challenging because many molecular diagnostic laboratories are running sequencing tests for tens to hundreds of target genes. As such, we aimed to design and implement a method-based proficiency testing (PT) method for Sanger sequencing and to assess its feasibility in Korea.
Methods:A pathogenic low density lipoprotein receptor (LDLR) variant was chosen as the positive PT material, and material without an LDLR variant was used as the negative PT material. We distributed the two PT materials with primer pair sets to 17 molecular diagnostic laboratories nationwide.We calculated the correct results (%) for variation type, zygosity, nucleotide change, amino acid change, variant classification, and clinical interpretation.
Results:Fourteen laboratories responded to the survey. The results for the two PT materials were 100% correct for all evaluation points including variant type, zygosity, nucleotide change, amino acid change, variant classification, and clinical interpretation.
Conclusions:This pilot PT survey demonstrates a feasibility of using method-based PT for assessing the Sanger sequencing performance of molecular diagnostic laboratories in Korea.
