슬라이드 1

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1 ARCHITECT i2000 를이용한 HOMOCYSTEINE 정량분석평가 짂단검사의학과생화학파트이유희

2 순서 Ⅰ. 서론 1) Homocysteine 의이해및소개 2) 시약교체배경 Ⅱ. 본론 1) 재료및방법 2) 결과 Ⅲ. 고찰

3 I. 서론

4 Homocysteine 메티오닌의이화작용 (catabolism) 에의해생성되는황을함유하는아미노산 혈장호모시스테인의수치는다음에영향을받는다. 유전적요인 (homocystinuria) 영양적요인 ( 결핍 ) Vitamin B6 (pyridoxal phosphate) Vitamin B12 (cobalamin) Folate 신장요인 SH CH 2 CH 2 + H 3 N CH C O O - Homocysteine 호모시스테인은자가산화를통해 homocystine (HCY-HCY), mixed disulfides (ie, protein-hcy, cysteine-hcy), homocysteine thiolactone 등을생성하며, 추가적으로환원된형태로도순환 => 이들을통틀어 total HCY 라고한다.

호모시스테인이시스테인으로변환시주효소로 cystath-ionine-ß-synthase (CBS), 조효소로 Vitamin B6 가필요 4.

5 Homocysteine 의대사과정 과정 1. 섭취한음식물의단백질에서분해된아미노산중메티오닌으로전환 2. 메티오닌은메틸기가분리되면서호모시스테인으로전환 3. 호모시스테인이시스테인으로변환시주효소로 cystath-ionine-ß-synthase (CBS), 조효소로 Vitamin B6 가필요 4. 호모시스테인은다시메티오닌으로변환주효소는 methionine synthase (MS), 조효소로 Vitamin B12 와엽산 (Folate) 필요 5. 위의대사과정은정상인에서이루어지며, 이과정에필요한조효소가부족하면, 대사과정이깨지면서호모시스테인이과량축적됨

6 Homocysteine 이왜위험한가? Plaque 형성, 성장 손상부위에지방성 물질축척 HCY 이동맥내 벽손상

7 Homocysteine 검사의임상적유용성 As an aid for screening patients suspected of having an inherited disorder of methionine metabolism including: - Cystathionine beta-synthase deficiency (Homocystinuria) - Methylenetetrahydrofolate reductase deficiency (MTHFR) and its thermolabile variants: - Methionine synthase deficiency - Cobalamin (Cbl) Metabolism: - Combined Methyl-Cbl and Adenosyl-Cbl deficiencies: Cbl C2, Cbl D2 and Cbl F3 deficiencies - Methyl-Cbl specific deficiencies: Cbl D-Var1, Cbl E and Cbl G deficiencies - Transcobalamin II deficiency - Adenosylhomocysteinase: AHCY deficiency - Glycine N-methyltransferase: GNMT deficiency - Methionine Adenosyltransferase I/III Deficiency: MAT I/III deficiency Evaluating patients for suspected deficiency of vitamin B12 or folate As a indicator of cardiovascular risk

8 Homocysteine 수치 : Normal and Elevated Normal levels of thcy: mol/l Elevated levels: 고호모시스테인혈증 (hyperhomocysteinemia) 만일하나또는그이상의 HCY 대사과정이억제 ( 비타민결핍, 효소결함 ) => HCY 축척 => 혈장에서의 thcy의수치증가 고호모시스테인혈증의수치 Desirable: 10 mol/l Intermediate: 10 to <15 mol/l High: 15 to <30 mol/l Very High: 30 mol/l

9 TAT 단축효과 시약교체배경 - Hitachi 7600 P module (open 시약 ) -> 다른검사 (lipase, Free fatty acid ) 결과에영향을미쳐분주후모아서검사실시 -> Homocysteine 검사후 special wash, pooled serum으로전처리 probe, cuvette 세척하는번거로움. - Archetect ( 전용시약 ) - 일회용 cuvette 사용 > carry over 방지 -> 실시간검사가능 시약단가절감 - Hitachi ,000원 /100T/kit -> Archetect 550,000원 /100T/kit 10% 절감효과검사건수 600건 / 월 ( 년간 6,480,000 원비용절감 ) - 사용배경 - (Advia centaur 로사용 )-> 장비 out ->open 시약으로 Hitachi 7600 에 setting -> Architect 도입후재검토 -> 검사이전

10 Hitachi 7600 전처리전, 후검사결과비교 (carry over 의영향 ) 시행전 NO Homocysteine Lipase Free Fatty Acid sample 시행후 NO Homocysteine Lipase Free Fatty Acid sample special wash ( probe, cuvette 세척 ), pooled serum 전처리

11 Homocysteine(HCY) 검사법비교 변경전 변경후 검사장비 Hitachi 7600 P module Architect i2000 검사방법 EIA (Enzyme Immuno Assay) CMIA (Chemiluminescent Microparticle Immuno Assay) 정상참고구간 mol/l mol/l 측정민감도 <0.5 mol/l <1.0 mol/l 측정범위 mol/l mol/l 결과보고시간 18 분 29 분 검체용량 16.5 μl 78 μl 자동화여부 x o

12 II. 본론

13 Homocysteine on Architect i2000

14 재료및방법 Precision test 일시 : ~ (5 일간 ) Material : low, medium,high level dedicated QC 물질 Comparison test 일시 : ~ Material : low, medium & high level 40 검체 Method : EIA vs. CMIA Linearity test 일시 : Material : CMIA Concentration (2.74) 와 (25.59) 인검체로 5 Level 4:0, 3:1, 2:2, 1:3, 0:4 비율로 3 회반복검사

15 Precision test 5 runs 4 repeats Level Total Mean(μmol/L) SD within (μmol/l) CV within(%) SD total (μmol/l) CV Total(%) Package Target(μmol/L) Low Medium High Reagent Lot No. (10171UP00), Calibrator Lot No. ( ), Control Lot No. ( )

16 Comparison test Sample Hitachi 7600 Architect i N= 40, R²=0.990

17 Comparison test Sample Hitachi 7600 Architect i

18 외부정도관리 (RIQAS) 2012 년 Hitachi 7600 Architect Mean 1 월 월 월 월 월 월 월 월 월 월 *RIQAS : Randox International Quality Assessment

19 Architect i2000 Architect Mean Passing Bablock 을이용한 Homocysteine 비교 X Hitachi 7600 Y Architect i2000 Architect= 0.88x Bias 95% CI Constant to Proportional to 0.91 X Hitachi 7600 Y Architect Mean Architect= 0.79x Bias 95% CI Constant to 2.28 Proportional to Scatter Plot with Passing & Bablok Fit 50 Scatter Plot with Passing & Bablok Fit Hitachi 7600 Identity Passing & Bablok (I) fit ( x) Hitachi 7600 Identity Passing & Bablok (I) fit ( x) 본원 Comparision 비교 외부정도관리비교

20 Evaluation of the Linearity 측정치와계산치로구한직선의회귀방정식은 y = 1.027x 이었고 R² 는 이었다.

21 III. 고찰 Architect i2000 을이용한 Homocysteine 검사의재현성우수 Hitachi 7600 과의상관성양호 TAT 단축효과와업무효율성증대 시약의단가절감

22 감사합니다

4. (034) 원저-김광수(17~23).hwp

4. (034) 원저-김광수(17~23).hwp Original Articles Journal of Korean Epilepsy Society Journal of Korean Epilepsy Society 14(1):17-23, ISSN 1226-6965 Received June 1, Accepted June 3, 고신대학교의과대학신경과학교실 Relationship of Folate, Vitamin B 12,