ARCHITECT i2000 를이용한 HOMOCYSTEINE 정량분석평가 짂단검사의학과생화학파트이유희
순서 Ⅰ. 서론 1) Homocysteine 의이해및소개 2) 시약교체배경 Ⅱ. 본론 1) 재료및방법 2) 결과 Ⅲ. 고찰
I. 서론
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.
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
Homocysteine 이왜위험한가? Plaque 형성, 성장 손상부위에지방성 물질축척 HCY 이동맥내 벽손상
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 http://www.mayomedicallaboratories.com/test-catalog/clinical+and+interpretive/80379
Homocysteine 수치 : Normal and Elevated Normal levels of thcy: 5.0-15.0 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
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 7600 640,000원 /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
Hitachi 7600 전처리전, 후검사결과비교 (carry over 의영향 ) 시행전 2012.08.17 NO Homocysteine Lipase Free Fatty Acid sample 1 15.1 122 446 2 17.5 84 297 3 16.2 150 707 4 9.8 123 403 5 10.4 90 948 6 9.6 92 196 7 11.5 96 1036 8 11.8 96 690 9 10 95 859 10 7 97 357 시행후 NO Homocysteine Lipase Free Fatty Acid sample 1 20 451 2 48 299 3 16 714 4 42 426 5 special wash 28 682 6 ( probe, cuvette 세척 ), 29 201 7 pooled serum 전처리 39 369 8 27 694 9 18 864 10 24 358
Homocysteine(HCY) 검사법비교 변경전 변경후 검사장비 Hitachi 7600 P module Architect i2000 검사방법 EIA (Enzyme Immuno Assay) CMIA (Chemiluminescent Microparticle Immuno Assay) 정상참고구간 5.0-15.0 mol/l 5.0-15.0 mol/l 측정민감도 <0.5 mol/l <1.0 mol/l 측정범위 1.0-46.0 mol/l 1.00 50.00 mol/l 결과보고시간 18 분 29 분 검체용량 16.5 μl 78 μl 자동화여부 x o
II. 본론
Homocysteine on Architect i2000
재료및방법 Precision test 일시 : 2011.08.06 ~ 2011.08.10 (5 일간 ) Material : low, medium,high level dedicated QC 물질 Comparison test 일시 : 2012.08.27 ~ 2012.09.07 Material : low, medium & high level 40 검체 Method : EIA vs. CMIA Linearity test 일시 : 2012.09.19 Material : CMIA Concentration (2.74) 와 (25.59) 인검체로 5 Level 4:0, 3:1, 2:2, 1:3, 0:4 비율로 3 회반복검사
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 7.01 0.27 3.89 0.26 3.76 7.56 Medium 12.70 0.37 2.93 0.36 2.85 13.66 High 25.72 0.78 3.03 0.72 2.81 27.15 Reagent Lot No. (10171UP00), Calibrator Lot No. (802887350), Control Lot No. (802888137)
Comparison test Sample Hitachi 7600 Architect i2000 1 5.70 4.14 2 4.10 3.74 3 6.80 5.10 4 6.60 5.02 5 4.80 4.03 6 6.50 4.72 7 3.70 2.82 8 10.50 8.63 9 3.90 3.16 10 31.30 26.32 11 29.80 24.77 12 30.20 26.87 13 28.20 26.52 14 24.00 21.19 15 25.30 20.13 16 22.30 19.85 17 23.60 21.57 18 26.40 23.11 19 20.10 17.14 20 23.60 19.36 21 25.60 20.80 22 30.40 26.86 23 24.10 20.33 24 7.50 6.26 25 13.00 11.06 26 12.90 10.71 27 7.60 6.52 28 8.20 6.71 29 12.30 9.22 30 11.00 8.60 31 8.70 5.90 32 14.80 11.54 33 11.30 9.92 34 10.60 8.61 35 14.40 11.31 36 10.50 8.27 37 13.80 11.52 38 14.00 12.18 39 13.10 9.90 40 13.20 10.33 N= 40, R²=0.990
Comparison test Sample Hitachi 7600 Architect i2000 1 14.00 12.18 2 14.40 11.31 3 14.80 11.54 4 22.30 19.85 5 23.60 21.57 6 23.60 19.36 7 24.00 21.19 8 24.10 20.33 9 25.30 20.13 10 25.60 20.80 11 26.40 23.11 12 28.20 26.52 13 29.80 24.77 14 30.20 26.87 15 30.40 26.86 16 31.30 26.32
외부정도관리 (RIQAS) 2012 년 Hitachi 7600 Architect Mean 1 월 24.4 20.612 2 월 6.0 5.435 3 월 46.1 35.967 4 월 7.3 5.973 5 월 22.2 18.502 6 월 35.0 29.398 7 월 5.1 5.376 8 월 42.9 34.934 9 월 23.2 19.416 10 월 35.6 27.694 *RIQAS : Randox International Quality Assessment
Architect i2000 Architect Mean Passing Bablock 을이용한 Homocysteine 비교 X Hitachi 7600 Y Architect i2000 Architect= 0.88x - 0.66 Bias 95% CI Constant -0.66-1.07to -0.17 Proportional 0.88 0.84to 0.91 X Hitachi 7600 Y Architect Mean Architect= 0.79x - 1.08 Bias 95% CI Constant 1.08 0.14to 2.28 Proportional 0.79 0.73to 0.83 35 Scatter Plot with Passing & Bablok Fit 50 Scatter Plot with Passing & Bablok Fit 30 45 40 25 35 20 15 10 5 0 0 10 20 30 Hitachi 7600 Identity Passing & Bablok (I) fit (-0.66 + 0.88x) 30 25 20 15 10 5 5 25 45 Hitachi 7600 Identity Passing & Bablok (I) fit (1.08 + 0.79x) 본원 Comparision 비교 외부정도관리비교
Evaluation of the Linearity 측정치와계산치로구한직선의회귀방정식은 y = 1.027x + 0.165 이었고 R² 는 0.999 이었다.
III. 고찰 Architect i2000 을이용한 Homocysteine 검사의재현성우수 Hitachi 7600 과의상관성양호 TAT 단축효과와업무효율성증대 시약의단가절감
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