Lab Med Qual Assur.  2025 Mar;47(1):28-31. 10.15263/jlmqa.2025.47.1.28.

Method-Based Proficiency Test Program for Assessing Quality of Sanger Sequencing-Based Molecular Tests

• Seong MW
• Sung Hb
• Lee JS

• KMID: 2566245
• DOI: http://doi.org/10.15263/jlmqa.2025.47.1.28

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.

Keyword

Sequence analysis; Labroatry proficiency testing; Method-based labroatry proficiency testing