임앙병리검매과약외 711 : 깨 133 권깨 12Q, 119-128, 21. 일반화학검샤용 DiaSys 시약의평가 춤북대학교병원임상병리과 분당재생병원임상병리과 ι 김천대학임상병리과 ** 황석연 홍승복장철수 ** Evaluation of DiaSYSCfD Reagents for Routine Cheical Tests Hwang, Seock Yeon., Hong, Sellllg Bok*., α1illlg, αleoul Soo** Departent Clini cul Pathol., Chungb μk Nat'l Univ. Hospital, Cheon와 ;u, Korea Departent Clinical Pathol., Bundang ]eseng. Hospital. Sungna, Korea* Departent qf Ci lnial Pathol.. Kicheoη Health Siences College, Kicheon, Korea** We evaluated the recently introduced DiaSys μ (DiaSys Diagnostic systes Inteational, Gennany) reagents for the routine cheical tests and copared with the reagents used in Chllllgbuk National University Hospital. We analyzed 15 ites for the evaluation of precisions, linearities and recovery rates. The coparison study between the current reagents used in Chllllgbuk National University Hospital and DiaSys reagents were also perfored for those ites. The linearities were good in all ites, and the recoverγ rates were acceptable results. The coefficients of variation (CV) of within-run and total precision were less than 5% in all ites. The correlation analysis revealed good results in ost ites, except calciu showing the correlation coefficient of.9551. DiaSys reagents for the routine cheical tests showed good precisions, linearities, recovery rates and correlations with the currently used reagents. We conclude that these reagents are suitable to the substitute for the currently used reagents. Key words DiaSys reagents, Routine cheical tests, Evaluation 1. 서를 저l 성등을고려하여다양한종류의시약이이용 되고였다. 사약의교체사 겸사실에서논반드시 일반화학겸사는임상검사설에서가장많은 겸사플하는부분으로검사결과의선뢰성뿐만 아니라시약의안정성, 조제의편리성그리고정 기존에사용하고있던시약파충분한평가환사 행한후시약을사용하는것이선뢰성있는결 파블얻는데중요하며 (David and Theodore, 1999), 119
이근 빼l 져 도미근 얘 떼 띈새로운분석방법에대한평가항목의종류와분 평가하였다 (NCCLS, 1986). 석기술에대한세부사항의기준이제시되어있 다 (Buttner et al., 1976; NCCLS, 1986; NCCLS, 4. 회수율평가 1992; NCCLS, 1995). 저자틀은국내에처음소개되는 DiaSys R. (Dia 회수율의평가는저농도와고농도의겸처 1 로 Sys Diagnostic Systes Inteational, Gerany) 시 3:1, 1:1, 1:3 의비율로섞어 4 회씩반복측정하여 약을기존에충북대학교병원에서사용하고있는 회수율을계산하였다. 사약과비교하여위 시약의효용성을평가하였 기에보고하는바이다. 5. 정밀도평가 n. 재료및방법 정상농도와고농도의 DiaSys 정도관리물질을 이용하여 167~ 의동일검체로나누어 -2 C 에냉 1. 시약 은뱀이반었U 하1f 학O--: 분사D 버ψ좌L땅얹O} 야기던분용검 二 %i 기않 메석 은A/ 어 야 으상호 --펴 벼b LM 와o U바o시 험셰 B 판d 민M 라과 이t 외 L E 근하 A 동츠기봐각서근제동보관하면서 16 일간연속하여 DiaSys 시약으 로측정하였다. 각군의검체는 1 얼 2 회 검사하 였으며각검사사에두번씩반복측정하여검사져도/때LU 빠새; 1,l P n 끼M 근,다근一r 디괴고사야빼V 미/.j 이잉E χ펴l1 여다, 3 껴L 사뼈자 φb총j 빠 미내도F 이O? 목별로본원에서사용하는기존의사약과측정원리는 Table 1 과같다. 6. 상관성평가 저농도, 정상농도그리고고농도의겸제가각 2. 대상 각 27~ 씩포함되도록하여총 6 명의환자겸제 2αU년 5-6윌사이에충북대학교병원임상병리과에의뢰된환자검제를대상으로각검사항목별로 6명의겸제를선택하여비교하였으며, 를대상으로가존에사용하고있던시약과 DiaSys 시약으로각각측정하여상관관계륜조사하였다 (NCCLS, 1995). 정밀도분석은 DiaSys 자제정도관리물질플이용하였다. 한편, 검사항목은일반화학검사항목. 결과 중에서 15 항목윤선택하여겸사를사행하였으며, 검사가기로논 autoated cheical analyzer <;>l 직선성평가 Hitachi 715 (Hitachi Inc., Japan) 을사용하였다. 총 15 종목에대하여저 농도와고농도검체틀 3. 직선성평가 4:, 3: 1, 2:2, 1 :3, :4 의비율로섞어만는겸체로 시행한직션성검정에서모두양호한결과룰보 제조사에서제시하는직선성의범위를고려하 였다 (Table 2). 여저농도와고농도의점제를선택하고각단계별로희석하여 5 가지농도의겸처1 틀만들어 2. 회수율평가 각각에대하여 4 회씩반복측정하여직선성을 총 15 종목에대하여시행한회수율은 95.85% 12
에서 17.81 % 로모두양호한결과를보였다 (Table 3). 3. 정밀도평가 검사내 (wit 비 n-run), 검사얼간 (between-day) 그 리고총정말도는변이계수 (CV) 가정상농도와 비정상농도의 DiaSys 정도관리물질에서 5% 이내 로양호하였다 (Table 4, 5). 16 일간 DiaSys 자체정도관리물질중정상농 도와고농도의정도관리물질을사용하여 I 거 asys 시약의정밀도를평가하였는데모두변이계수가 5% 이내로양호하였으며 (Table 4, 5), DiaSys 전 용정도관리물질이외에 Bio-Rad (Bio-Rad, Ltd., France), Roche (Roche, Ltd., USA) 그리고 Merk 4. 상관성평가 (Merk, Gennany) 등의정도관리물질들을사용하 여 동열한시약으로측정하였는데이들도대부 총 15 종목에대하여기존의사약과비교한 분이변이계수가 5% 이내로양호한결과를보였 관성평가에서감숨 (r=.9551) 을저l 외한상관계 다 수 (r) 는.98 이상으로우수한상관성을보였다. 상관성검사를위하여기존의시약으로저농 그리고 Bland-Altan 방법 (1 986) 에의한각종목도, 중간농도그리고고농도의겸체가각각 27R 별차이의평균과표준편차그리고 95% 선뢰구 간 (confidence interval) 을계산하였는데대체로 씩포함되도록총 6 겸제를선택하여 DiaSys 시 약과동시에측정한결과감숨 (r=.9551) 을제외 양호한 일치의한계 "( 1 빼 ts of agreeent"; ean 한대부분의종목에서상관계수는.9872 이상으 i: 2SD) 를보였다 (Table 6, Fig. 1). 로매우양호한결과를보였으며, 감숨, 무기인, 편, 직선성검정에사용하였던저농도와고농도 의검체를 3:1, 1:1 그리고 1:3 의비율로섞어측 정한값음계산한값과비교한회수율도모든 종목에서양호하였다 (Table 3). N. 고화임상검사실에서시약의안정성, 조제의편리성그라고경제성등을고려하여다양한종류의 시약을사용하게되는데, 시약의안정성및정확 성등이검사결과에마치는영향은매우크므 로겸사에사용되는시약이바뀔때마다정확한 시약의평가가요구된다하겠다 (David and Theodore, 1999). 최근에국내에처음으로소개되는 DiaSys 사약을충북대학교병원에서기존에사용 하고었는시약들과비교하였기에보고하는바 이다. 제조사에서제시하는직선성의범위릎고려하 여기존의시약으로저농도와고농도의검체를 선택하여사행한직선성검정에서직선의결정계수 (r 2 ) 는모두.99 이상이였으며직선관계식 의기울기및 y 전편도양호하였다 (Table 2). 한 를총단백그리고알부민항목은비교범위가좁아정확한상관성을반영하지못할수있으므로추후보다넓은범위의평가가필요할것으로생각된다 (Table 6, Fig. 1). 캄숨항목의경우상관계수가.9551 이었으며상관방정식의기울기는.886 이었다. 그리고 Bland-Altan의방법에의한두시약간차이는 DiaSys 시약이기존의시약보다평균.1367 g/dl이낮았고이들차이의표준편차논.446 이었으며, 총분석오차 (total anal ytical eπor) 는 CLIA 88 에서제시한세개의결정농도 (decision level 7., 1.8, 13. 아ng/dL) 중 13. g/dl에서만허용범위를벗어났다 (CLIA, 1992). 이결과의정확한원언을밝히지는못하였지만보다넓은범위와많은검체그리고참고물질 (reference aterial) 과의비교평가로정확성을평가해야될것으로사료되며, 실제로이항목을검사실에적용하기위해서는검사실자 체의참고범위를재설정해야할것으로판단 121
된다. ALP와 GGT 의경우에상관방정식의기울기가.912와.888로비율오차 (proportional erro디를보였으나상관계수는.9992와.9975 로양호하였으며비교범위도충분히넓고두검사항목에서계산한총분석오차 (total analytical error) 는 CLIA 88 에서저1 시한최대허용총오차범위내에속하였으며, 또한, ALP, GGT 이외의종목에서도총분석오차는모두 CLIA 88 에서제시한허용범위에속하였다 (CLIA, 1992). 그리고 Bland-Altan 방법에의한시약간차이의평균과표준편차는 Table 6 에제시하였는데이들의 일치의한계 "( liits of agreeent" ean i: 2SD) 논 DiaSys시약으로대체사용시선뢰할수 있을정도로충분히작았다 (Bland and Altan, 1986). 간섭물질은분석물질을측정하는데영향을 u] 치는불리적영향과효소와같은물질의측정시화학적영향등을임으켜총분석오차의주요한원인이될수있다 (NCCLS, 1986). 실제로 IFCC 나 NCCLS 등에서는임상검사실에서새로운검사방법을선택할때약물, 혈색소, 빌리루빈, 지절그리고단백질등외부및내부인자의간섭효과를평가하도록권고하고있다 (Galteau and Siest, 1984). 그러나본연구에서는비교항목이많아간섭물질의영향을평가하지는못하였으며추후이들의영향에대한평가가이루어져야할 Table 1. Coparison of analytical principles between DiaSys and current reagent Current reagents Paraeter Copany Principles of ethods DiaSys reagents Calciu Wako pure cheical Co., Japan Colorietry (OCPC) Colorietry (OCPC) Phosphorus Wako pure cheical Co., Japan Phosphoolybdate, UV Phosphoolybdate, UV Total protein In Hwa Phar. Co., Korea Biuret Biuret Albuin In Hwa Phar. Co., Korea BCG BCG Glucose Asan Phar. Co., Korea HK, G6PD, UV HK, G6PD, UV BUN Daiichi Cheical Co., Japan Urease-Indophen 이 (Berthelot) Urease with GLDH Creatinine Daiichi Cheical Co., Japan Jaffe, kinetic Jaffe, kinetic Uric acid Eiken Cheical Co., Japan Uricase, Colorietry Uricase, Colorietry AST Asan Phar. Co., Korea 37C IFCC (AACC) 37C IFCC (AACC) -P-5 -P -seru bla따 -P-5 -P -seru blank ALT Asan Phar. Co., Korea 37C IFCC (AACC) 37C IFCC (AACC) -P-5 -P -seru blank -P-5 -P -seru blank ALP Roche Diagnostics Co., Japan 37C P-NPP, AMP, 37C P-NPP, AMP, IFCC (Bower-McCob) IFCC (Bower-McCob) GGT Daiichi Cheical Co., Japan 37C IFCC (carboxyl- 37C IFCC (carboxylated-ggpna) ated-ggpna) Cholesterol Asan Phar. Co., Korea Enzyatic, Colorietry Enzyatic, Colorietry Triglyceride MBL, Ltd. Co., Korea Glycerol phosphate Glycerol phosphate oxidase (GPO) oxidase (GPO) Aylase Asan phar. Co., Korea B-G5-CNP B-G5-CNP 122
것으로생각된다. 시약의선택에서검사결과의선뢰성뿐아니 라시약조제의편리성그리고보관시안정성등 도고려해야하는데, DiaSys 시약은액상으로사 약의조저l 가필요없이바로사용할수있으므 로대량겸체를분석하는일반화학검사에적당 하며, 분말시약의조저 l 사벨 - 생할수있는오류 나오염의가능성윤판연수있윤것으로생각 된다. 결론적으로일반화학검사용 DiaSys 시약은 정빌도, 직선성및회수율이우수하였으며일부 종목에서비교범위가충분히넓지못하였지만 기존의상품화된시약과비교하여우수한상관 성을보였다. 그리고대부분의사약이액상이므 로시약사용의편리성파안정성측면에서임상 검사실에서사용하는데무리가없플것으로판 단되었다. V. 결로 저자들은최근에국내에처음소개되는마aSys 일반화학검사사용시약을기존에충북대학교병원에서사용하고있는시약과비교하여평가하고자하였다. 일반화학검사중자주검사하는 15 항목을선택하여정밀도, 직선성및회수율을분석하였다. 그리고기존에사용하고있던시약과상관성을비교하였다. 15 항목의모두에서정맏도, 직선성및회수율이양호하였으며, 감숨음제외하고는모든항목에서상관계수가.9872 이상으로우수하였다. DiaSys 일반화학검사용시약은정말도, 직선성및회수율이우수하였으며기존의시약과비교하여우수한상관관계틀보여기존사약의대제에무리가없을것으로사료된다. Table 2. Linearity test results of DiaSys reagents Paraeter Unit Range Slope Intercept Coefficient of deterination(r 2 ) Calciu g/dl 4.4-14.6 Phosphorus g/dl.28-16.8 Total protein g/dl 4.6-12.5 Albuin g/dl 2.4-5.4 Glucosε g/dl 46-445 BUN g/dl 7.3-77 Creatinine g/dl.5- l7.2 Uric acid g/dl 1.6-8.7 AST IUjL 2-171 ALT IUjL 2-181 ALP IUjL 53-799 GGT IUjL 8-691 Cholesterol g/dl 57-97 Triglyceride g/dl 56-49 Aylase IUjL 9-539 1.236 -.2153.996.995.142.999.9832.1418.998 1.373 -.1 792.998 1.15 -.8141.999 1.25.4469.998.9798.298.998.9976.246.999.9989 5.9866.999.9928 6.4613.999 1.2 -.484.999 1.97-1.6532.999.9938 7.5818.999.971 7.9781.995 1.16.7133.999 123
S 서g서gTable 3. Recovery rate results of DiaSys reagents Paraeter Calciu % h J M P rl 아외뼈떼삐U Glucose BUN nu l Creatinine Uric acid AST ALT ALP GGT Cholesterol Triglyceride 뻐SI 4.4 S2 14.6 3:1 Recovery rate(%) 1: 1 1:3 11.5 12.14.28 16.8.88 98 15.22 4.6 12.5 99.5.24 2.4 5.4.43.51 뻐46 445.58.82 뻐7.3 77 12.38 12.83.5 1.6 2 2 53 8 57 56 17.2 8.7 171 181 799 691 97 49 95.85 99.71 11. 12.31 99.47.9 99.7 96.5 98.41 11.57 11.31.51 98.42.36 12.45 17.22.22.6 14.l6 15.25 94.36 98.35 12.39 17.81 Aylase 9 538.12 1 1.37 13.21 S 1 = the saple of the lowest concentration S2= the saple of the highest concentration Table 4. Within-run, between-day and total precision of DiaSys reagents for DiaSys control aterials of noral concentrat1on Paraeter Calciu Phosphorus T. protein Alburnin Glucose BUN Creatinine Uric Acid AST ALT ALP GGT Cholesterol Triglyceride Aylase Unit 따 앤때 맨때 맨ιnι빠H 따까H H H 씨싸셰싸n께싸찌싸께싸n n n 빠께싸맺맺R 따#Mean 9.47 4.2 1 5.2 1 3.14 99.97 19.42 1.28 4.45 4.3 32.58 95.l9 47.78 17.4 118.l 98.1 Within-run pre 디 slon SD(Swr).l54 CV(%) 1.627.85 2.15.52 M.35 1. 119 1.639 1.64.375 1.931.22 1.692.92 2.85.75 1.873.76 2.33 1.392 1.462.968 2.26.829.772 1.639 1.388 2.66 2.714 SD(Sdd).245.64.65.6 1.689.441.39.77 l.41.797 2.636.523 l.933 2.357 3.22 Between-day preclslon CV(%) 2.592 1.59 1.248 1.916 1.69 2.271 3.9 1.737 2.61 2.449 2.769 1.95 l.8oo 1.997 3.285 Total precision SD(ST).28 CV(%) 2.962.12 2.843.86 1.646.72 2.275 2.21 2.211.565 2.91.45 3.52.l22 2.742 l.241 3.99 1.95 3.36 2.861 3.6 l.417 2.965 2.l54 2.6 2.838 2.42 4.228 4.31 124
Table 5. Within-run, between-day and total precision of DiaSys reagents for DiaSys control aterials of abnoral concentratlon. Within-run Between-day Paraeter Unit Mean preclslon preclslon Total precision SD(Swr) CV(%) SD(Sdd) CV(%) SD(ST) CV(%) Calciu gjdl 13.83.2 19 1.583.249 1.798.326 2.354 Phosphoflli g/dl 7.1.58.821.118 1.657.136 1.923 T. protein gjdl 5.2.34.652.89 1.711.94 1.87 Albuin gjdl 2.96.33 1. 11.53 1.791.63 2.114 Glucose g/dl 247.7 2.787 1. 125 3.86 1.246 3.958 1.598 BUN g/dl 66.42.8 1.25 1.324 1.993 1.532 2.36 Creatinine g/dl 3.37.38 1.18.18 3.194.1 17 3.462 Uric Acid g/dl 1.75.112 1.42.199 1.855.32 2.975 AST IU/L 146.4 1.262.862 2.76 1.418 2.335 1.595 ALT IU/L 133.9 1. 14.923 3.169 2.366 3.336 2.491 ALP IU/L 343.7 3.248.945 5.768 1.678 6.596 1.978 GGT IU/L 235.9 2.693 1.1 41 2.4 18 1.25 4.261 1.86 Cholesterol g/dl 23.1 2.14 1.53 4.279 2.16 4.838 2.38 Triglyceride gjdl 222.9 2.955 1.326 5.297 2.377 6.18 2.74 Aylase IU/L 26.9 2.781 1.66 2.883 1.15 5.161 1.978 Table 6. Coparison study between DiaSys reagents and currents reagents Paraeter Unit Range Correlation Bland-Altan bias[6] slope Mean(d)* SDt 95% CI t Calciu gjdl 5.5-1.9.886.9551 -.14.446 -.25 - -.3 Phosphorus gjdl 1.7-7.4.967.9873 -.15.41 -.25 - -.5 T. protein gjdl 4.9-9. 1.38.9872.16.199.1 1 -.21 Albuin g/dl 2.2-5.8 l.94.9849.9.239.3 -.15 Glucose gjdl 11-463 1.1.998 -.56 4.875-1.79 -.67 BUN gjdl 2-126 1.12.998.32 1.599 -.8 -.72 Creatinine gjdl.2-7..973.998.4.19.1 -.7 Uric acid g/dl.9-14.4.984.9982 -.14.234 -.2 - -.8 AST IU/L 4-31.99.999 -.3 2.686 -.71 -.65 ALT IU/L 4-29.989.9985-1.88 4.694-3.6 - -.69 ALP IU/L 17-443.912.9992-14.6 5.45-15.9 - -13.3 GGT IU/L 1-641.888.9975-1.53 17.22-14.9 - -6.17 Cholesterol gjdl 5-468.994.9953.8 1.467-2.57-2.73 Triglyceride gjdl 44-4 1.946.9973-4.28 8.236-6.36 - -2.2 Aylase IU/L 56-4 1.957.995-5.42. 9.732-7.88 - -2.96 Nuber of tests for each ites 6 * ean difference of current reagents fro diasys reagent concentration. stan없rd deviation of the difference of current reagents fro diasys reagent. 95% Confidence interval (ean 土 1.96 X SD/ιn, n=6) 125
> - A낙[] 끼/ CU O O Phosphorus y =.9672x -.63 r =.9873 j 싹? t w Wako. t 6 l-- 35 3 25 잊 2 픔 15 5 o o y =.99x +.6975 r = 잃 9 5 * * AST 15 2 Asan -- 25 3 35 T. protein ALT > -o 1 9 QU 7l [O 4 4 y = 1.77x -.18 r =.9872 i* :. 씬.*... ~. - 4 6 7 Inhwa 1 > 3 2 잉 2 15 5 O O y =.9894x -1.1256 r =.9985 --- - - - - - - * --- -- 一 - --- ---..-1 5 15 2 25 3 Asan Albuin 5,εε ALP 6 I y=1.맹4x -.2968 r =.9849 훌. 훌 옆 4{ 줬 i á5 4 I 디 3 I t!+.,.ii i I y =.9123x -.3381 4 I r =.9992._,. 잊 3 I $. 협 2 1.~ 깐In I-Mta -----------' 2 3 4 5 Roche 냐 Glucose y = 1.14x -.763. f. X 2 3 Asan 4 5 GGT Fig. 1. Coparison study between DiaSys reagents and currents reagents (continued) 126
15) (/) 디 5 BUN y = 1.121x -.167 r =.998 낸칸一. 5) 15 Daiichi c g@n: 5 4 2[ x pu Cholesterol r =.9953 ~..# -. -샤 2 3 4 5 Asan Creatinine Triglyceride y =.9728x +.854 r =.998 옳응 5 4 3C 2 =.9461 x + 4.4 845 r =.9973.,, -" Daiichi 2 3 4 5 MBL Aylase g}o j y = 9567x + 1 8839 4 r =.995 뿔 j /.. ~... - s -., -- 2 3 Asan 4 5 Fig. 2. Coparison study between DiaSys reagents and current reagents 잡고문흰 1. Bland JM and Altan DG. Statistical ethod for assessing agreeent between two ethods of clinical easureent. Lμncet, 1:37-31, 1986 2. Buttner J, Borth R, Boutwell JH, Broughton PMG and Bowyer RC. (lfcc Coittee on Standards): Provisional recoendation on quality control in clinical cheistηii. Assessent of Analytical Methods for routine use. Clin Che 127
22:1922-1932, 1976 3. Clinical Laboratory Iproveents Aendents of 188. Final Rule. Laboratory Requireents. Federal Resister 57:72-7288, 1992 4. David DK and Theodore P1. Selection and Evaluation of Methods. In: Burtis CA, Ashwood ER. Tietz textbook of clinical cheistη, pp. 32-335 3rd ed. Philadelphia: WB Saunders copany, 1999 5. G떠teau ~αi{ and Si않t G. 바ug effects in clinical cheistη, p따t 2. Guideline for evaluation of analytical interference. J Clin Chelη Bioche, 22:275-279, 1984 6. National Coittee for Clinical Laboratory Standards: Evaluation of the linearity of quantitative analytical ethods.proposed guideline. Docuent EP6-P. Villanova, PA, National coittee for Clinical Laboratory Standards, 1986 7. National Coittee for Clinical Laboratory Stan없rds: Interference testing in clincal cheistη. Proposed guidline. Docuent EP7-P. Villanova, National coittee for Clinical Laboratoη Standards, 1986 8. National Coittee for Clinical Laboratory Standards: Evaluation of precision perforance of clinical cheistry devices-second edition. Tentative guideline Docuent EP5-T2. Villanova, PA, National Coittee for Clinical Laboratory Standards, 1992 9. National Coittee for Clinical Laboratory Standards: Method coparison and bias estiation using patient saples. Approved guideline. Docuent EP9-A. Villanova, PA, National coittee for Clinical Laboratory Standards, 1995 128