• Sitemap
  • Contact us

pISSN 2950-9114 eISSN 2950-9122
Article View

Report On Proficiency Testing

J Lab Med Qual Assur 2018; 40(4): 199-210

Published online December 31, 2018

https://doi.org/10.15263/jlmqa.2018.40.4.199

Copyright © Korean Association of External Quality Assessment Service.

Report on the External Quality Assessment Scheme for Molecular Diagnostics in Korea (2017)

Man Jin Kim, Mi-Hye Yoon, Ji Yun Song, Sung Im Cho, Sung-Sup Park, Moon-Woo Seong

Department of Laboratory Medicine, Seoul National University Hospital, Seoul National University College of Medicine, Seoul, Korea

Correspondence to:Moon-Woo Seong Department of Laboratory Medicine, Seoul National University Hospital, 101 Daehak-ro, Jongno-gu, Seoul 03080, Korea Tel: +82-2-2072-4180 Fax: +82-2-747-0359 E-mail: MWSeong@snu.ac.kr

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Quality control for genetic analysis has become more important with a drastic increase in testing volume and clinical demands. The molecular diagnostics division of the Korean Association of Quality Assurance for Clinical Laboratory conducted two trials in 2017 on the basis of molecular diagnostics surveys, involving 53 laboratories. The molecular diagnostics surveys included 37 tests: gene rearrangement tests for leukemia (BCR-ABL1, PML-RARA, AML1-ETO, and TEL-AML1), genetic tests for Janus kinase 2, FMS-like tyrosine kinase 3-internal tandem duplication, FMS-like tyrosine kinase 3-tyrosine kinase domain, nucleophosmin, cancer-associated genes (KRAS, EGFR, KIT, and BRAF), hereditary breast and ovarian cancer genes (BRCA1 and BRCA2), Li-Fraumeni syndrome (TP53), Wilson disease (ATP7B), achondroplasia (FGFR3), hearing loss and deafness (GJB2), Avellino (TGFBI), multiple endocrine neoplasia 2 (RET), Huntington disease, spinocerebellar ataxia, spinal and bulbar muscular atrophy, mitochondrial encephalopathy with lactic acidosis and stroke-like episodes, myoclonic epilepsy ragged red fibre, Leber hereditary optic neuropathy, Prader-raderd Angelman syndrome, Duchenne muscular dystrophy, spinal muscular atrophy, fragile X syndrome, apolipoprotein E genotyping, methylenetetrahydrofolate reductase genotyping, and ABO genotyping. Molecular genetic surveys revealed excellent results for most participants. The external quality assessment program for genetic analysis in 2017 proved useful for continuous education and the evaluation of quality improvement.

Keywords: Laboratory proficiency testing, Molecular pathology, Molecular genetics

2017년도 분자진단검사 신빙도조사는 전체 회원기관에게 참가 신청조사를 하여 총 2회 시행하였고 회차별 37종목에 각 2종의 검체를 참여기관에 배포하여 다양한 돌연변이 및 유전질환을 포함하였다.

1. 분자진단검사

1차 신빙도조사는 2017년 4월 17일에 검체를 발송하고 2018년 1월 10일에 결과를 통보하였고, 2차 신빙도조사는 2017년 9월 19일에 검체를 발송하고 2018년 2월 10일에 결과를 통보하였다. 신빙도조사 물질의 회차별 검체와 검체번호, 검체구성은 Table 1과 같다.

Table 1 . Survey tests and specimens for external quality assessment of the molecular diagnostics test survey in 2017.

   TestsSpecimens   Test specimens of 1st trialSpecimens   Test specimens of 2nd trial
Leukemia (BCR-ABL1)GH 17-01Normal cell lineGH 17-11Leukemia cell line (BCR/ABL1 positive)
GH 17-02Normal cell lineGH 17-12Normal cell line
Leukemia (PML-RARA)GH 17-01Normal cell lineGH 17-11Normal cell line
GH 17-02Leukemia cell line (PML/RARA positive)GH 17-12Normal cell line
Leukemia (AML1-ETO)GH 17-01Normal cell lineGH 17-11Normal cell line
GH 17-02Normal cell lineGH 17-12Normal cell line
Leukemia (TEL-AML1)GH 17-01Leukemia cell line (TEL/AML1 positive)GH 17-11Normal cell line
GH 17-02Normal cell lineGH 17-12Leukemia cell line (TEL/AML1 positive)
JAK2GH 17-03Extracted DNA with JAK2 mutationGH 17-13Normal DNA
GH 17-04Normal DNAGH 17-14Extracted DNA with JAK2 mutation
FLT3-ITDGH 17-05Normal DNAGH 17-15Normal DNA
GH 17-06Extracted DNA with FLT3-ITD mutationGH 17-16Extracted DNA with FLT3-ITD mutation
FLT3-TKDGH 17-07Extracted DNA with FLT3-TKD mutationGH 17-17Extracted DNA with FLT3-TKD mutation
GH 17-08Normal DNAGH 17-18Normal DNA
NPM1GH 17-09Extracted DNA with NPM1 mutationGH 17-19Normal DNA
GH 17-10Normal DNAGH 17-20Extracted DNA with NPM1 mutation
KRASGS 17-01Extracted DNA with KRAS mutationGS 17-09Normal DNA
GS 17-02Normal DNAGS 17-10Extracted DNA with KRAS mutation
EGFRGS 17-03Extracted DNA with EGFR mutationGS 17-11Extracted DNA with EGFR mutation
GS 17-04Normal DNAGS 17-12Normal DNA
BRAFGS 17-05Normal DNAGS 17-13Extracted DNA with BRAF mutation
GS 17-06Extracted DNA with BRAF mutationGS 17-14Normal DNA
KITGS 17-07Normal DNAGS 17-15Normal DNA
GS 17-08Extracted DNA with KIT mutationGS 17-16Extracted DNA with KIT mutation
BRCA1GG 17-01Extracted DNA with BRCA1 mutationGG 17-41Normal DNA
GG 17-02Normal DNAGG 17-42Extracted DNA with BRCA1 mutation
BRCA2GG 17-03Normal DNAGG 17-43Extracted DNA with BRCA2 mutation
GG 17-04Extracted DNA with BRCA2 mutationGG 17-44Normal DNA
TP53GG 17-05Extracted DNA with TP53 mutationGG 17-45Normal DNA
GG 17-06Normal DNAGG 17-46Extracted DNA with TP53 mutation
ATP7BGG 17-07Extracted DNA with ATP7B mutationGG 17-47Extracted DNA with ATP7B mutation
GG 17-08Normal DNAGG 17-48Normal DNA
MELASGG 17-09Extracted DNA with MELAS mutationGG 17-49Normal DNA
GG 17-10Normal DNAGG 17-50Extracted DNA with MELAS mutation
MERRFGG 17-11Normal DNAGG 17-51Extracted DNA with MERRF mutation
GG 17-12Extracted DNA with MERRF mutationGG 17-52Normal DNA
GJB2GG 17-13Extracted DNA with GJB2 variantGG 17-53Extracted DNA with GJB2 variant
GG 17-14Normal DNAGG 17-54Normal DNA
LHONGG 17-15Extracted DNA with LHON mutationGG 17-55Extracted DNA with LHON mutation
GG 17-16Normal DNAGG 17-56Normal DNA
RETGG 17-17Extracted DNA with RET mutationGG 17-57Extracted DNA with RET mutation
GG 17-18Normal DNAGG 17-58Normal DNA
SCA1GG 17-19Normal DNAGG 17-59Normal DNA
GG 17-20Extracted DNA with SCA1 mutationGG 17-60Normal DNA
SCA2GG 17-19Normal DNAGG 17-59Normal DNA
GG 17-20Normal DNAGG 17-60Normal DNA
SCA3GG 17-19Normal DNAGG 17-59Normal DNA
GG 17-20Normal DNAGG 17-60Normal DNA
SCA6GG 17-19Normal DNAGG 17-59Extracted DNA with SCA6 mutation
GG 17-20Normal DNAGG 17-60Normal DNA
SCA7GG 17-19Normal DNAGG 17-59Normal DNA
GG 17-20Normal DNAGG 17-60Normal DNA
APOE genotypeGG 17-21Extracted DNA with APOE ε3/ε3 genotypeGG 17-61Extracted DNA with APOE ε4/ε4 genotype
GG 17-22Extracted DNA with APOE ε4/ε4 genotypeGG 17-62Extracted DNA with APOE ε3/ε4 genotype
AchondroplasiaGG 17-23Normal DNAGG 17-63Extracted DNA with FGFR3 mutation
GG 17-24Extracted DNA with FGFR3 mutationGG 17-64Normal DNA
MTHFR genotypeGG 17-25Extracted DNA with MTHFR 677T/T homozygoteGG 17-65Extracted DNA with MTHFR 677C/T heterozygous
GG 17-26Normal DNAGG 17-66Extracted DNA with MTHFR 677T/T homozygote
PWS/ASGG 17-27Normal DNAGG 17-67Extracted DNA with PWS mutation
GG 17-28Extracted DNA with AS mutationGG 17-68Normal DNA
DMDGG 17-29Normal DNAGG 17-69Extracted DNA with DMD mutation
GG 17-30Extracted DNA with DMD mutationGG 17-70Normal DNA
HDGG 17-31Extracted DNA with HD mutationGG 17-71Extracted DNA with HD mutation
GG 17-32Normal DNAGG 17-72Normal DNA
FMR1GG 17-33Normal DNAGG 17-73Normal DNA
GG 17-34Extracted DNA with FMR1 mutationGG 17-74Extracted DNA with FMR1 mutation
TGFBIGG 17-35Extracted DNA with TGFBI mutationGG 17-75Normal DNA
GG 17-36Normal DNAGG 17-76Extracted DNA with TGFBI mutation
SBMAGG 17-37Normal DNAGG 17-77Normal DNA
GG 17-38Extracted DNA with AR mutationGG 17-78Extracted DNA with AR mutation
SMAGG 17-39Extracted DNA with SMN1 mutationGG 17-79Extracted DNA with SMN1 mutation
GG 17-40Normal DNAGG 17-80Normal DNA
ABO genotypeGO 17-01Extracted DNA with cis-AB/B genotypeGO 17-03Extracted DNA with B/O genotype
GO 17-02Extracted DNA with A/O genotypeGO 17-04Extracted DNA with cis-AB/A genotype

Abbreviations: JAK2, Janus kinase 2; FLT3, FMS-like tyrosine kinase 3; ITD, internal tandem duplication; TKD, tyrosine kinase domain; NPM1, nucleophosmin; EFGR, epidermal growth factor receptor; MELAS, mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes; MERRF, myoclonic epilepsy with ragged red fibres; LHON, Leber hereditary optic neuropathy; RET, multiple endocrine neoplasia 2; SCA, spinocerebellar ataxia; APOE, apolipoprotein E; MTHFR, methylenetetrahydrofolate reductase; PWS/AS, Prader-Willi/Angelman syndrome; DMD, Duchenne muscular dystrophy; HD, Huntington disease; FMR1, fragile X mental retardation-1; TGFBI, transforming growth factor beta induced; SBMA, spinal and bulbar muscular atrophy; SMA, spinal muscular atrophy..


각 기관의 보고에 따라 검사방법, 검사결과 및 검사방법별 검사결과를 비교 분석하였으며 기관별 보고결과에 대하여 기본적인 평가(acceptable, unacceptable)를 실시하였다. 합격/불합격의 기준은 10기관 미만의 참여항목의 경우 주관기관의 검증결과와 일치하는 경우 acceptable이라고 판정하였으며 10기관 이상의 참여항목의 경우 주관기관의 검증결과와 참여기관 중 80% 이상의 일치가 있을 경우 acceptable이라고 판정하였다. 삼염기 반복 유전질환인 Huntington disease (HD), spinocerebellar ataxia (SCA1, SCA2, SCA3, SCA6, SCA7), spinal and bulbar muscular atrophy (SBMA), fragile X syndrome (FMR1) 검사의 정량값은 평가에 포함시키지 않았다.

1. 분자진단검사

1) 참여기관 및 검사항목

2017년 1차 및 2차 신빙도조사에 참여한 기관은 53개 기관이었다. 분자진단검사는 혈액종양 8종목, 고형종양 4종목, 선천성 유전질환 8종목, 미토콘드리아 유전질환 3종, 삼염기 반복 유전질환 7종, 신경근육 유전질환 1종, 기타 유전질환 3종, 유전형검사 3종목 등 총 37종목에 대해 시행하였다.

혈액종양 분자진단검사 8종목인 leukemia and lymphomas (BCR-ABL1, PML-RARA, AML1-ETO, TEL-AML1), JAK2, FMS-like tyrosine kinase 3 (FLT3-internal tandem duplication [ITD], FLT3-tyrosine kinase domain [TKD]), nucleophosmin (NPM1) 분자진단검사는 1차에서 각각 36기관, 34기관, 33기관, 31기관, 30기관, 19기관, 17기관, 16기관이, 2차에서 37기관, 36기관, 35기관, 33기관, 31기관, 20기관, 18기관, 18기관이 참가하였다. 고형종양 분자진단검사 4종목인 KRAS, BRAF, EGFR, KIT 분자진단검사는 1차에서 10기관, 15기관, 9기관, 10기관이 참가하였고, 2차에서는 11기관, 14기관, 10기관, 10기관이 참가하였다. 유전질환 8종목인 BRCA1, BRCA2, TP53, ATP7B, FGFR3, GJB2, TGFBI, RET 분자진단검사는 1차에서 각각 16기관, 16기관, 6기관, 8기관, 9기관, 10기관, 13기관, 7기관이 참가하였으며, 2차에서 15기관, 15기관, 6기관, 8기관, 9기관, 10기관, 13기관, 7기관이 참가하였다. 삼염기 반복 유전질환과 신경근육 유전질환 종목인 HD, SCA1, SCA2, SCA3, SCA6, SCA7, SBMA, FMR1 분자진단검사에는 1차에서 각각 5기관, 6기관, 7기관, 6기관, 6기관, 6기관, 5기관, 10기관이 참가하였고, 2차에서 5기관, 6기관, 7기관, 6기관, 6기관, 6기관, 5기관, 10기관이 참가하였다. 미토콘드리아 유전질환에 속하는 종목인 mitochondrial encephalopathy with lactic acidosis and stroke like episodes (MELAS), myoclonic epilepsy ragged red fibre (MERRF), Leber hereditary optic neuropathy (LHON₋major mutation) 검사에는 1차에서 각각 6기관, 5기관, 6기관이 참가하였고, 2차에서 6기관, 5기관, 6기관이 각각 참가하였다. Prader-Willi/Angelman syndrome (PWS/AS), Duchenne muscular dystrophy (DMD), spinal muscular atrophy (SMA) 분자진단검사는 1차에서 각각 5기관, 6기관, 6기관이 각각 참가하였고, 2차에서 5기관, 6기관, 6기관이 각각 참가하였다. 유전형검사에 속하는 종목인 apolipoprotein E (APOE) genotyping, methylenetetrahydro-folate reductase (MTHFR) genotyping, ABO genotyping 분자진단검사는 1차에서 각각 32기관, 13기관, 13기관이 참가하였으며, 2차에서 34기관, 14기관, 13기관이 참가하였다.

2) 검사방법

각 기관별 사용하는 검사법은 Table 2와 같다.

Table 2 . Mutation detection methods of the molecular diagnostics test survey in 2017.

   Tests Mutation detection methods (lab-developed test)ResponseMutation detection methods (commercial kit)Response
Leukemia (BCR-ABL1)Allele-specific PCR1Hemavision29
PCR (include multiplex), gel electrophoresis1
PCR and gel electrophoresis2
Real-time PCR4
Leukemia (PML-RARA)Allele-specific PCR1Hemavision29
PCR (include multiplex), gel electrophoresis1
PCR and gel electrophoresis2
Real-time PCR3
Leukemia (AML1-ETO)Allele-specific PCR1Hemavision29
PCR (include multiplex), gel electrophoresis1
PCR and gel electrophoresis2
Real-time PCR2
Leukemia (TEL-AML1)Allele-specific PCR1Hemavision29
PCR (include multiplex), gel electrophoresis1
PCR and gel electrophoresis2
JAK2Allele-specific PCR7
Pyrosequencing1
Real-time PCR10
Real-time PCR, hydrolysis probe2
Sanger sequencing10
Single base extension (SNaPshot)1
FLT3-ITDAllele-specific PCR1
PCR and fluorescent based capillary electrophoresis (fragment length analysis)3
PCR (include multiplex), gel electrophoresis1
PCR and gel electrophoresis1
RFLP1
Sanger sequencing12
FLT3-TKDAllele-specific PCR1
PCR and fluorescent based capillary electrophoresis (fragment length analysis)1
PCR (include multiplex), gel electrophoresis1
PCR and gel electrophoresis10
Sanger sequencing2
Other*2
NPM1Real-time PCR1
RFLP1
Sanger sequencing16
KRASCodon12-pyrosequencing2
Codon13-pyrosequencing2
Codon61-pyrosequencing2
Codon12-Sanger sequencing8
Codon13-Sanger sequencing8
Codon61-Sanger sequencing8
BRAFcodon 600 (p.V600E)-pyrosequencing2
codon 600 (p.V600E)-real-time PCR6
codon 600 (p.V600E)-real-time PCR, hydrolysis probe3
codon 600 (p.V600E)-Sanger sequencing4
codon 600 (p.V600K)-pyrosequencing2
codon 600 (p.V600K)-Sanger sequencing4
codon 600 (p.V600K)-pyrosequencing2
codon 600 (p.V600K)-Sanger sequencing4
EGFRCodon719-pyrosequencing1
Codon790-pyrosequencing1
Codon858-pyrosequencing1
Codon861-pyrosequencing1
Exon 19 deletion-pyrosequencing1
Codon719-Sanger sequencing8
Codon790-Sanger sequencing8
Codon858-Sanger sequencing8
Codon861-Sanger sequencing8
Exon 19 deletion-Sanger sequencing8
KITcodon 816-real-time PCR, hydrolysis probe1
codon 816-Sanger sequencing8
codon 816-other1
Exon 9-Sanger sequencing6
Exon 9-other1
Exon 11-Sanger sequencing6
Exon 11-other1
Exon 13-Sanger sequencing6
Exon 13-other1
Exon 17-Sanger sequencing7
Exon 17-other1
BRCA1Sanger sequencing16
BRCA2Sanger sequencing16
TP53Sanger sequencing6
ATP7BSanger sequencing8
AchondroplasiaSanger sequencing9
GJB2Sanger sequencing10
TGFBIReal-time PCR3
Sanger sequencing10
RETSanger sequencing7
HDPCR and fluorescent based capillary electrophoresis (fragment length analysis)5
SCA1PCR and fluorescent based capillary electrophoresis (fragment length analysis)6
SCA2PCR and fluorescent based capillary electrophoresis (fragment length analysis)7
SCA3PCR and fluorescent based capillary electrophoresis (fragment length analysis)6
SCA6PCR and fluorescent based capillary electrophoresis (fragment length analysis)6
SCA7PCR and fluorescent based capillary electrophoresis (fragment length analysis)6
SBMAPCR and fluorescent based capillary electrophoresis (fragment length analysis)5
LHONSanger sequencing5
Single base extension (SNaPshot)1
MELASSanger sequencing6
MERRFSanger sequencing5
PWS/ASPCR and gel electrophoresis, methylation specific5
DMDMLPA6
SMAMLPA4
RFLP2
APOE genotypeAllele-specific PCR5
PCR (include multiplex), gel electrophoresis3
PCR and gel electrophoresis1
Real-time PCR17
Real-time PCR, hydrolysis probe1
Real-time PCR, melting curve analysis1
RFLP1
Sanger sequencing2
Line probe assay1
MTHFR genotypeAllele-specific PCR2
PCR and gel electrophoresis1
Real-time PCR1
Real-time PCR, hydrolysis probe1
Real-time PCR, melting curve analysis1
RFLP4
Sanger sequencing2
Single base extension (SNaPshot)1
ABO genotypeAllele-specific PCR1
RFLP1
Sanger sequencing9
Single base extension (SNaPshot)2
FMR1PCR and fluorescent based capillary electrophoresis (fragment length analysis)6
Southern blot1
Triplet repeat primed PCR and fluorescent based capillary electrophoresis2

Abbreviations: PCR, polymerase chain reaction; JAK2, Janus kinase 2; FLT3, FMS-like tyrosine kinase 3; ITD, internal tandem duplication; RFLP, restriction fragment length polymorphism; TKD, tyrosine kinase domain; NPM1, nucleophosmin; EFGR, epidermal growth factor receptor; RET, multiple endocrine neoplasia 2; HD, Huntington disease; SCA, spinocerebellar ataxia; SBMA, spinal and bulbar muscular atrophy; LHON, Leber hereditary optic neuropathy; MELAS, mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes; MERRF, myoclonic epilepsy with ragged red fibres; PWS/AS, Prader-Willi/Angelman syndrome; DMD, Duchenne muscular dystrophy; MLPA, multiple ligation probe amplification; SMA, spinal muscular atrophy; APOE, apolipoprotein E; MTHFR, methylenetetrahydrofolate reductase; FMR1, fragile X mental retardation-1..

*No response.


3) 신빙도조사 결과

Leukemia and lymphomas 분자진단검사는 1차에서 TEL-AML1 재배열과 PML-RARA 재배열 검체를, 2차에서는 TEL-AML1 재배열과 BCR-ABL1 재배열 검체로 구성하였다. 1차 평가에서 PML-RARA, AML1-ETO 재배열 문항에 대하여 참여한 모든 기관이 예상결과와 일치하는 결과를 보고하였고, BCR-ABL1 재배열 문항은 1개 기관이 다른 subtype을 보고하였다. TEL-AML1 재배열 문항은 31개 기관 중 1개 기관이 예상결과와 상이한 결과를 보고하였다. 2차에서는 BCR-ABL1, PML-RARA, AML1-ETO 재배열 문항에 대하여 참여한 모든 기관이 예상결과와 일치하는 결과를 보고하였고 TEL-AML1 재배열 문항은 33개 기관 중 2개 기관이 예상결과와 상이한 결과를 보고하였다.

JAK2, FLT3-ITD, FLT3-TKD, NPM1 분자진단검사에서는 JAK2, NPM1에 참여한 모든 기관이 1차와 2차에서 예상결과와 일치하는 결과를 보고하였다. FLT3-ITD 분자진단검사에서는 1차에 참여한 모든 기관이 예상결과와 일치하는 결과를 보고하였으나 2차에서는 1개 기관이 정상 검체에서 변이를 발견해 보고하였으며 FLT3-TKD 분자진단검사에서는 1차에서 1개 기관이 변이를 발견하지 못하였으며 2차에서는 전체 응답결과의 80% 이상의 consensus를 보이지 않아 전체 기관의 결과를 “ungraded”로 보고하였다.

KRAS, BRAF, EGFR, KIT 분자진단검사에서는 KRAS, BRAF, EGFR 분자진단검사에 참여한 모든 기관이 1차와 2차에서 예상결과와 일치하는 결과를 보고하였다. KIT 분자진단검사에서는 1차에서 1개 기관이 예상결과와 상이한 결과를 보고하였고 2차에서는 모든 기관이 예상결과와 일치하는 결과를 보고하였다.

BRCA1, BRCA2, TP53, ATP7B, GJB2, FGFR3, TGFBI, RET 분자진단검사에서는 BRCA2, TP53, FGFR3, TGFBI에 참여한 모든 기관이 1차와 2차에서 예상결과와 일치하는 결과를 보고하였다. BRCA1 분자진단검사에서는 2차에 참여기관 중 1개 기관이 예상결과와 다른 염기변이를 보고하였다. ATP7B 분자진단검사에서는 1차에 참여한 1개 기관이 예상 변이부위와 상이한 부위의 변이를 보고하였으나 2차에서는 예상결과와 일치하는 결과를 보고하였다. GJB2 분자진단검사에서는 1차와 2차에 참여한 각 1개 기관이 예상 변이부위와 상이한 부위의 변이를 보고하여 평가에서 제외되었다. RET 분자진단검사에서는 1차에서 예상결과와 일치하는 결과를 보고하였으나 2차에서는 1개 기관이 예상결과와 다른 단백질변이를 보고하였다.

HD, SCA1, SCA2, SCA3, SCA6, SCA7, SBMA 분자진단검사에서는 SCA1, SCA2, SCA3, SCA6, SCA7 분자진단검사에 참여한 모든 기관이 예상결과와 일치하는 결과를 보고하였다. HD, SBMA 분자진단검사에서는 2차에 참여한 기관 중 각각 2개 기관이 예상결과와 상이한 결과를 보고하였다.

MELAS, MERRF, LHON 분자진단검사에서는 MERRF, LHON 분자진단검사에 참여한 모든 기관에서 1차와 2차에서 예상결과와 일치하는 결과를 보고하였다. MELAS 분자진단검사에서는 1차에서 1개 기관이 예상결과와 상이한 결과를 보고하였으나 2차에서는 참여한 모든 기관에서 예상결과와 일치하는 결과를 보고하였다. PWS/AS, DMD, SMA 분자진단검사에서는 참여한 모든 기관이 1차와 2차에서 예상결과와 일치하는 결과를 보고하였다.

APOE, MTHFR, ABO genotyping 분자진단검사에서는 MTHFR genotyping 분자진단검사에 참여한 모든 기관이 1차와 2차에서 예상결과와 일치하는 결과를 보고하였다. APOE genotyping 분자진단검사에서는 1차에 제공한 1번 검체에 대하여 1개 기관이 예상결과와 상이한 결과를 보고하였다. ABO genotyping 분자진단검사에서는 1차에 제공한 1번 검체에 대하여 2개 기관이 예상결과와 상이한 결과를 보고하였으며 2번 검체에 대하여 1개 기관이 예상결과와 상이한 결과를 보고하였다.

FMR1 분자진단검사에서는 1차에 참여한 1개 기관이 평가용 검체로 제공한 2종류의 검체 모두에서 예상결과와 다른 결과를 보고하였으나 2차에서는 참여한 모든 기관이 예상결과와 일치하는 결과를 보고하였다. 각 기관별 보고결과를 정리한 정오답 통계는 Table 3과 같다.

Table 3 . Results of the molecular diagnostics test survey carried out in 2017.

   TestsSpecimens of 1st trial   Intended responsesTotal no. of participants/acceptable responseSpecimens of 2nd trial   Intended responsesTotal no. of participants/acceptable response
Leukemia (BCR-ABL1)GH 17-01Mutation not detected36/36GH 17-11BCR/ABL1 positive37/37
GH 17-02Mutation not detected36/36GH 17-12Mutation not detected37/37
Leukemia (PML-RARA)GH 17-01Mutation not detected34/34GH 17-11Mutation not detected36/36
GH 17-02PML/RARA positive34/34GH 17-12Mutation not detected36/36
Leukemia (AML1-ETO)GH 17-01Mutation not detected33/33GH 17-11Mutation not detected35/35
GH 17-02Mutation not detected33/33GH 17-12Mutation not detected35/35
Leukemia (TEL-AML1)GH 17-01TEL/AML1 positive31/30GH 17-11Mutation not detected33/33
GH 17-02Mutation not detected31/31GH 17-12TEL/AML1 positive33/31
JAK2GH 17-03p.Val617Phe mutation30/30GH 17-13Mutation not detected31/31
GH 17-04Mutation not detected30/30GH 17-14p.Val617Phe mutation31/31
FLT3-ITDGH 17-05Mutation not detected19/19GH 17-15Mutation not detected20/19
GH 17-06FLT3-ITD mutation19/19GH 17-16FLT3-ITD mutation20/20
FLT3-TKDGH 17-07FLT3- TKD mutation17/16GH 17-17FLT3-TKD mutation18/all ungraded
GH 17-08Mutation not detected17/17GH 17-18Mutation not detected18/18
NPM1GH 17-09NPM1 mutation16/16GH 17-19Mutation not detected18/18
GH 17-10Mutation not detected16/16GH 17-20NPM1 mutation18/18
KRASGS 17-01Codon 12 mutation10/10GS 17-09Mutation not detected11/11
GS 17-02Mutation not detected10/10GS 17-10Codon 12 mutation11/11
EGFRGS 17-03Exon 19 deletion9/9GS 17-11Exon 19 deletion10/10
GS 17-04Mutation not detected9/9GS 17-12EGFR mutation10/10
BRAFGS 17-05Mutation not detected15/15GS 17-13p.Val600Glu14/14
GS 17-06p.Val600Glu15/15GS 17-14Mutation not detected14/14
KITGS 17-07Mutation not detected10/10GS 17-15Mutation not detected10/10
GS 17-08Codon 816 mutation10/9GS 17-16Codon 816 Mutation12/12
BRCA1GG 17-01c.5074+1G>T, heterozygote16/16GG 17-41Mutation not detected15/15
GG 17-02Mutation not detected16/16GG 17-42c.3442delG, p.Glu1148Argfs*7, heterozygote15/14
BRCA2GG 17-03Mutation not detected16/16GG 17-43Mutation not detected15/15
GG 17-04c.8140C>T, p.Gln2714*, heterozygote16/16GG 17-44c.7480C>T, p.Arg2494*, heterozygote15/15
TP53GG 17-05c.742C>T, p.Arg248Trp, heterozygote6/6GG 17-45Mutation not detected6/6
GG 17-06Mutation not detected6/6GG 17-46c.742C>T, p.Arg248Trp, heterozygote6/6
ATP7BGG 17-07c.3809A>G, p.Asn1270Ser, heterozygote c.3247C>T, p.Leu1083Phe, heterozygote8/7GG 17-47c.3104G>T, p.Gly1035Val, heterozygotec.3247C>T, p.Leu1083Phe, heterozygote8/8
GG 17-08Mutation not detected8/8GG 17-48Mutation not detected8/8
MELASGG 17-09m.3243A>G6/5GG 17-49Mutation not detected6/6
GG 17-10Mutation not detected6/6GG 17-50m.3243A>G6/6
MERRFGG 17-11Mutation not detected5/5GG 17-51m.8344A>G5/5
GG 17-12m.8344A>G5/5GG 17-52Mutation not detected5/5
GJB2GG 17-13c.35delG, p.Gly12Valfs*, heterozygote10/9GG 17-53c.35delG, p.Gly12Valfs*, heterozygote10/10
GG 17-14Mutation not detected10/10GG 17-54Mutation not detected10/10
LHONGG 17-15m.11778G>A6/6GG 17-55Mutation not detected6/6
GG 17-16Mutation not detected6/6GG 17-56m.11778G>A6/6
RETGG 17-17c.2753T>C, p.Met918Thr, heterozygote7/7GG 17-57c.1859G>C, p.Cys620Ser, heterozygote7/6
GG 17-18Mutation not detected7/7GG 17-58Mutation not detected7/7
SCA1GG 17-19Mutation not detected6/6GG 17-59Mutation not detected6/6
GG 17-20SCA1 mutation6/6GG 17-60Mutation not detected6/6
SCA2GG 17-19Mutation not detected7/7GG 17-59Mutation not detected7/7
GG 17-20Mutation not detected7/7GG 17-60Mutation not detected7/7
SCA3GG 17-19Mutation not detected6/6GG 17-59Mutation not detected6/6
GG 17-20Mutation not detected6/6GG 17-60Mutation not detected6/6
SCA6GG 17-19Mutation not detected6/6GG 17-59SCA6 mutation6/6
GG 17-20Mutation not detected6/6GG 17-60Mutation not detected6/6
SCA7GG 17-19Mutation not detected6/6GG 17-59Mutation not detected6/6
GG 17-20Mutation not detected6/6GG 17-60Mutation not detected6/6
APOE genotypeGG 17-21APOE ε3/ε3 genotype32/31GG 17-61APOE ε4/ε4 genotype34/34
GG 17-22APOE ε4/ε4 genotype32/31GG 17-62APOE ε3/ε4 genotype34/34
AchondroplasiaGG 17-23Mutation not detected9/9GG 17-63p.Gly380Arg9/9
GG 17-24p.Gly380Arg9/9GG 17-64Mutation not detected9/9
MTHFR genotypeGG 17-25MTHFR 677T/T homozygous genotype13/13GG 17-65MTHFR 677C/T heterozygous genotype14/14
GG 17-26Mutation not detected13/13GG 17-66MTHFR 677T/T homozygous genotype14/14
PWS/ASGG 17-27Mutation not detected5/5GG 17-67Prader-Willi syndrome5/5
GG 17-28Angelman syndrome5/5GG 17-68Mutation not detected5/5
DMDGG 17-29Mutation not detected6/6GG 17-69Exon 56-57 deletion6/6
GG 17-30Exon 18-44 deletion6/6GG 17-70Mutation not detected6/6
HDGG 17-31Full penetrance5/5GG 17-71Full penetrance5/5
GG 17-32Mutation not detected5/5GG 17-72Mutation not detected5/3
FMR1GG 17-33Full mutation10/9GG 17-73Mutation not detected10/10
GG 17-34Mutation not detected10/9GG 17-74Full mutation10/10
TGFBIGG 17-35p.Arg124His13/13GG 17-75Mutation not detected13/13
GG 17-36Mutation not detected13/13GG 17-76p.Arg124His13/13
SBMAGG 17-37Mutation not detected5/5GG 17-77Full penetrance5/3
GG 17-38Full penetrance5/5GG 17-78Mutation not detected5/5
SMAGG 17-39SMN1, Exon7, homozygous deletion6/6GG 17-79SMN1, Exon7, homozygous deletion6/6
GG 17-40Mutation not detected6/6GG 17-80Mutation not detected6/6
ABO genotypeGO 17-01cis-AB/B genotype13/11GO 17-03B/O genotype13/13
GO 17-02A/O genotype13/12GO 17-04cis-AB/A genotype13/13

Abbreviations: FLT3, FMS-like tyrosine kinase 3; ITD, internal tandem duplication; TKD, tyrosine kinase domain; NPM1, nucleophosmin; EFGR, epidermal growth factor receptor; MELAS, mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes; MERRF, myoclonic epilepsy with ragged red fibres; LHON, Leber hereditary optic neuropathy; RET, multiple endocrine neoplasia 2; SCA, spinocerebellar ataxia; APOE, apolipoprotein E; MTHFR, methylenetetrahydrofolate reductase; PWS/AS, Prader-Willi/Angelman syndrome; DMD, Duchenne muscular dystrophy; HD, Huntington’s disease; FMR1, fragile X mental retardation-1; TGFBI, transforming growth factor beta induced; SBMA, spinal and bulbar muscular atrophy; SMA, spinal muscular atrophy..


2017년도 분자진단검사 신빙도조사는 전체적으로 매우 우수한 결과를 보였다[1-4]. BCR-ABL1, PML-RARA, AML1-ETO 재배열 문항, JAK2, NPM1, KRAS, BRAF, EGFR, BRCA1, BRCA2, TP53, FGFR3, TGFBI, SCA1, SCA2, SCA3, SCA6, SCA7, MERRF, LHON PWS/AS, DMD, SMA, MTHFR genotyping의 항목은 참여한 모든 기관이 1차와 2차에서 예상결과와 일치한 결과를 보고하였다.

검사범위가 한정된 KIT를 사용하는 기관에서는 검사부위 이외의 부위에서도 변이를 검출할 수 있는 대안을 마련하길 권장한다. 검체가 바뀌어 분석되었다고 예상되는 기관이 있어왔으므로 해당 기관에서는 매 회차마다 분석과정을 반드시 확인해야 한다.

HGVS에서는 오류를 방지하는 차원에서 triple-letter amino acid code 사용을 권장하고 있으므로 모든 참여기관들에서는 protein level에서의 변이에 대해 triple-letter amino acid code를 사용한 결과입력을 권장하며 특히 유전자변이 명명에서 잘못된 변이부위를 보고하거나 단백질변이 오기가 많이 발생하므로 최종보고 전 확인할 수 있는 체계를 구성하는 것을 권장한다.

동일 유전자를 검사하는 여러 기관에서 각기 다른 reference transcript를 사용하는 경우가 있으므로 검사 담당자는 주기적으로 reference transcript를 확인해야 한다. 변이에 따른 해석보고와 분석한 정량값으로 유전질환을 진단하는 경우 변이 해석과 정량값의 기준이 변하는 경우가 종종 있으므로 관련 문헌 및 참고자료를 주기적으로 확인해야 하며 HGVS 및 ACMG 관련 지침을 준수하는 것을 권장한다[5-7].

  1. Kim SH, Ki CS, Kim S, Kwon MJ, Kim JW, Park SS, et al. Annual report on external quality assessment in diagnostic genetics in Korea (2009). J Lab Med Qual Assur 2010;32:147-70.
  2. Kim SH, Ki CS, Jeon BR, Lee ST, Yoo EH, Kim JW, et al. Annual report on external quality assessment in diagnostic genetics in Korea (2008). J Lab Med Qual Assur 2009;31:161-81.
  3. Kim SH, Ki CS, Kim JH, Oh SH, Kim JW, Park SS, et al. Annual report on external quality assessment in diagnostic genetics in Korea (2007). J Lab Med Qual Assur 2008;30:167-87.
  4. Kim SH, Kim JW, Ki CS, Kim IS, Nam MH, Cho HJ, et al. Annual report on external quality assessment in diagnostic genetics in Korea (2005). J Lab Med Qual Assur 2006;28:153-67.
  5. Shaffer LG, McGowan-Jordan J, Schmid M. International Standing Committee on Human Cytogenetic Nomenclature. ISCN 2013: an international system for human cytogenetic nomenclature (2013). Basel: S. Karger, 2013.
  6. Richards CS, Bale S, Bellissimo DB, Das S, Grody WW, Hegde MR, et al. ACMG recommendations for standards for interpretation and reporting of sequence variations: revisions 2007. Genet Med 2008;10:294-300.
    Pubmed CrossRef
  7. Zhang VW, Wang J. Determination of the clinical significance of an unclassified variant. Methods Mol Biol 2012;837:337-48.
    Pubmed CrossRef

Supplementary File


Share this article on :

Stats or metrics