용혈이 각종 생화학 검사결과에 미치는 영향

홍숙희,김용호 인제대학교 1981 仁濟醫學 Vol.2 No.3

화학검사의 정확성과 결과 해석에 영향을 미치는 요인은 에러가지가 있지만 그중 응혈은 여러가지 이유에서 중요한 의의를 가진다. 저자들은 현재 부산 백병원 임상병리과에서 일상 행해지고 있는 각종 화학검사법에 대해 용혈이 미치는 영향을 관찰하기위해 혈청내 유리 혈색소를 점차 증가시키면서 검사 결과치와의 상호관계를 정량적으로 관찰하였든 바, 대부분의 화학검사가 용혈의 심한 정도에 비례하여 영향을 받고 있고 그중에서도 효소종목이 가장 큰 변화를 보여 LDH, acid phosphatase, SGOT는 정상치를 벗어나 몇 배의 증가를 보이고 있다. 아주 소량의 용혈이 있어도 각종 검사는 비용혈 혈청에 비해 통계적으로도 유의한 변화를 보여 용혈에 의한 영향은 명백하므로 용혈이 있는 검체는 그 결과해석에 신중을 기해야 한다. Hemolysis is a common cause of unsatisfactory serum samples obtained for clinical chemistry analysis, because it affects the accuracy of the chemical determination and the interpretation of the laboratory results. We have studied the hemolytic effects on various chemistry values quantitavely, determined in Busan Paik Hospital Laboratory everday, by serially increasing serum flee hemoglobin concentration on nonhemolyzed control pooled serum. The results obtained are as fellows: 1.Most of the chemistry values are affected significantly by hemolysis, in direct proportion to the degree of hemolysis. 2.Among the chemical determinations, enzyme groups are most affected by hemolysis. Electolytes except for potassium are least affected and nonenzyme groups are noticeably affected by hemolysis. 3.Of the enzymes, LDH appears to be most sensitive to hemolysis and the total increase of 1485.4U/ml results in 4.6 folds higher than nonhemolyzed control serum. GOT, acid phosphatase, alkaline phosphatase, GPT, and Gamma-GTP are order of increasing rate by hemolysis. 4.Of the electolytes, potassium is most sensitive to hemolysis and shows an increase of 69.7% compared with nonhemolyzed control serum. Lesser effects are observed in inorganic phosphate, chloride and calcium. 5.Of the non-enzyme groups, BUN is markedly increased by hemolysis. Total protein and triglyceride are also significantly increased but glucose is rather decreased. 6.Several chemistry values, enzymes in particular, potassium, and non-enzymes, show significant changes even though in trace hemolysis, and we should dive careful attention in the interpretation of the results obtained from hemolyzed serum.

신장이식 환자에서의 ABO 부(副)-부적합에 의한 용혈성 빈혈

이동렬 ( Dong Ryeol Lee ),강화미 ( Hwa Mi Kang ),김민웅 ( Min Woong Kim ),김치훈 ( Chi Heun Kim ),박종환 ( Jong Hwan Park ),윤지훈 ( Ji Hoon Yoon ),공진민 ( Jin Min Kong ) 대한내과학회 2005 대한내과학회지 Vol.69 No.2

저자들의 경우 ABO 부 부적합 이식 환자 중 ABO항체에 의한 용혈성 빈혈의 발생 빈도가 9%로 비교적 높은데 이는 공여자 특이 수혈(DST)을 시행한 것과 관련이 있을 수 있다. 또한 Azathiopurin, MMF와 같은 B임파구의 증식을 억제하는 약제의 사용이 적었던 것도 빈도의 증가에 기여했을 수 있다. ABO 부(副) 부적합에 의한 신 이식의 경우 초기 원인 불명의 빈혈이 관찰될 때 급성 용혈성 빈혈을 조기에 확인하고 감별 진단하는 것이 필요하며, 적절한 치료를 시행하면 대부분의 경우에서 이식 신 기능 장애 없이 회복이 가능하다. Background : Immune hemolysis secondary to ABO minor incompatibility is a rare graft versus host disease in renal recipients, secondary to anti-ABO antibody produced by lymphocytes of donor origin that reacts against recipient RBCs. Methods : To investigate the incidence and clinical features of immune hemolysis secondary to ABO minor incompatibility in renal allograft recipients, clinical records of 358 renal transplantation performed in Maryknoll Hospital since 1991 were analyzed retrospectively. Results : Fifty four (15%) of 358 renal transplants were ABO minor incompatible. Immune hemolysis secondary to anti-ABO antibody developed in 5 (9.2%) of 54 ABO minor incompatible renal transplant recipients. Immune hemolysis occurred in 3 (13.6%) patients among 22 allografts from blood type O donor to A recipients and 2 (10%) patients among 20 from blood type O donor to B recipients. All 5 patients received cyclosporin with prednisolone, and MMF was administered to one patient additionally. Immune hemolysis developed on 14±3 days after renal transplantation and lasted for about 10±3 days. The maximum reduction of hemoglobin was 3.3±1.0g/dL. All patients required donor type (blood type O) washed RBCs transfusion (5.0±2.6 units per patient) and plasmapheresis were performed in 3 patients (4.0±1.0 per patient). All patients recovered without deterioration of graft function. Age, number of HLA mismatch, creatinine at 1 year after transplantation, frequency of acute rejection and serum cyclosporin level during first 2 weeks were not significantly different between hemolysis group (N=5) and non-hemolysis group (N=49). Living unrelated transplantation is associated with increased incidence of immune hemolysis compared with living related transplantation (p<0.01). Conclusion : Although immune hemolysis secondary to ABO minor incompatibility is uncommon, we experienced cases with marked reduction of hemoglobin that required a large amount of transfusion. Therefore, this type of immune hemolysis needs to be considered as a differential diagnosis of posttransplant hemolysis. As our center routinely performs donor specific transfusion (DST), the incidence may be higher than that of other centers where DST is not usually given.(Korean J Med 69:177-182, 2005)

Interpretation of Folate Results in Hemolytic Plasma Samples: A Practical Approach

Minović Isidor,Dikkeschei Lambert D.,Vos Michel J.,Kootstra-Ros Jenny E. 대한진단검사의학회 2021 Annals of Laboratory Medicine Vol.41 No.5

Folate analysis in plasma is affected by hemolysis, which can lead to biased results. However, the degree of hemolysis that is considered acceptable is unclear. We explored the relationship between folate concentration and degree of hemolysis. Heparin plasma samples (N=77, hemolysis index ≤10 μmol/L) were spiked with increasing amounts of corresponding patient-specific hemolysate. Subsequently, the folate concentration and hemolysis index were measured using two Roche Cobas platforms, and their incremental relationship was investigated. The folate concentration ranged from 2.9 to 30.9 nmol/L with a median (interquartile range) of 11.4 (8.6–19.1) nmol/L. The linear relationship between the increments in folate concentration and hemolysis index was approximated by the function y=1.86x+1.56 (R2=0.996), where x represents the laboratory-specific critical difference in folate concentration, which can be calculated from the analytical variation of the employed folate assay(s), and y represents the hemolysis threshold. The hemolysis threshold did not significantly differ between the tertiles of plasma folate concentration (P=0.10). In conclusion, we have provided an evidence-based approach that can be used to reliably interpret folate concentrations in hemolytic samples, independent of the patient’s folate status.

12주간의 배드민턴 훈련이 나타내는 적혈구 용혈의 형태학적변화와 근 손상지표의 변화

박계순,전태원,소위영 대한임상건강증진학회 2009 Korean Journal of Health Promotion Vol.9 No.1

Background Red blood cell plays an important role in the function of transporting, providing oxygen to tissues, releasing CO2 from the body, and protecting cells from hydrogen ions. However, hemolysis caused by exercise may effect anemia, the reduction of hemoglobin, and VO2max. Furthermore, this hemolysis disrupts homeostasis. Previous studies on hemolysis have concentrated mainly on elite athletes and subjects having taken part in high intensity exercise and little is known about the effect of hemolysis from long term exercise. The purpose of this study was to investigate changes to hemolysis when subjected to long term exercise, badminton. Methods Thirty-five healthy, middle-aged women volunteered for this study and agreed to play badminton for one hour a day at 60∼70% intensity of their Heart Rate Reserve four times per week for 12 weeks. Blood samples were collected pre-exercise, immediately post-exercise, and recovery one hour from the antecubital vein before-training and after-training. Results Red blood cells, white blood cells, creatine phosphokinase (CPK), and lactate dehydrogenase (LDH) increased significantly; and these changes continued after the 12-week training period. Hemoglobin and hematocrit did not change with each exercise session nor after the 12-week training period. Erythrocyte sedimentation rate (ESR), peripheral blood smear (PBS) morphology, and ammonia increased significantly after a single exercise session. However, these changes were not detected after the 12-week training period. Conclusions CPK and LDH did not decrease but PBS morphology, ESR, and ammonia did. An increase in the latter three tests are considered to be direct indicators of hemolysis. These results demonstrate that the magnitude of hemolysis was reduced by the 12-weeks of exercise training. 연구배경 적혈구의 기능은 신체 조직에 충분한 산소를 공급하고, 조직으로부터 이산화탄소를 추출하여 신체의 외부로 배출하는 기능을 하며, 수소이온농도의 변화를 감지하여 인체를 방어하는 화학적 완충제로서의 역할을 담당한다. 그렇기 때문에, 운동으로 인한 적혈구의 용혈은 운동성 빈혈뿐만 아니라 헤모글로빈의 감소와 함께 최대산소섭취량과 운동 중 유산소 능력에 부정적인 영향을 미칠 수 있으며, 인체의 항상성을 위협할 수도 있다. 적혈구 용혈에 관한 지금까지의 선행연구들은 운동선수들이나 비교적 고강도의 운동에 참가한 사람들을 대상으로 일회성 운동에 국한되어 관찰하였다. 따라서 본 연구는 종전 연구에 한걸음 정진하여 장기간의 훈련이 적혈구 용혈 현상에 어떠한 영향을 미치는지를 규명하고자 하는 데에 그 목적이 있다. 방 법 본 연구의 피험자들은 35명으로 구성되었으며, 12주간 최대여유심박수(HRR: Heart Rate Reserve)의 60∼70%, 주4회, 60분으로 구성된 배드민턴 단식 게임을 진행하였다. 장기간의 훈련에 따른 적혈구 용혈 현상의 변화를 살펴보기 위한 혈액체취는 배드민턴 훈련 프로그램이 시작되는 1주차와 훈련 프로그램이 종료되는 12주차 때 운동전, 운동후, 회복기 1시간 후에 상완정맥에서 채혈하였다. 결 과 적혈구, 백혈구, 크레아틴 인산효소, 젖산탙수소효소는 훈련 1주차 때 증가현상을 나타내었으며, 훈련 12주차 때에도 증가현상을 나타내었다. 헤모글로빈, 헤마토크리트치는 훈련 1주차, 12주차 때 모두 증가현상을 나타내지 않았다. 적혈구 침강속도, 적혈구 형태학적 변화, 암모니아는 훈련 1주차 때 증가현상을 나타내었으나, 훈련 12주차 때에는 감소현상이 나타났다. 결 론 본 연구결과를 통하여 12주간의 배드민턴 훈련은 적혈구 용혈 현상의 직접적인 지표인 적혈구 침강속도, 적혈구 형태학적 변화, 암모니아의 감소를 관찰할 수 있었기에, 장기간의 규칙적인 훈련은 적혈구 용혈 현상을 완화시킬 수 있음을 나타내었다.

화학검사 결과의 검증을 위한 용혈 지수 일치화 도구 개발

표상신,남현수,차영종,이승관,이혜경 대한임상검사과학회 2017 대한임상검사과학회지(KJCLS) Vol.49 No.4

The hemolysis index (HI) is semi-quantitative marker for hemolysis. Because the characteristics of the HI vary from one commercial platform to another, no standardization or harmonization of the HI is currently available. Specimens (N=40) randomly selected from clinical patients were artificially hemolyzed in vitro. The serum of the specimens was then diluted with a 20 mg/dL difference between 0∼300 mg/dL based on serum hemoglobin measured using the XE-2100 hematology automation equipment (Sysmex, Japan). Diluted serum was measured using the Hitachi-7600 biochemical automation equipment (Hitachi, Japan) to differentiate between HI and serum hemoglobin. The data showed linearity between HI and serum hemoglobin and that HI 1 contained approximately 20 mg/dL of serum hemoglobin. To determine the blood rejection threshold, the HI was divided into three groups: HI 0∼1, HI 4∼6, HI 9∼15. After another batch of clinical specimens (N=40) was measured using a Hitachi-7600 (Hitachi, Japan), each specimen was moved forward and backward with the piston of the syringe to induce an artificial in vitro hemolysis, then measured again with a Hitachi-7600 (Hitachi, Japan). The percentage difference between the three groups was analyzed by ANOVA or the Kruskal-Wallis test. In the post-test, there were significant differences between the HI 0∼1 and the HI 5∼6: Glucose, creatinine, total protein, AST, direct bilirubin, uric acid, phosphorus, triglyceride, LDH, CPK, Magnesium, and potassium levels. Because many clinical tests differed significantly, the threshold for hemolysis could be appropriate for HI 5 (serum hemoglobin 100 mg/dL). 용혈 지수(hemolysis index)는 생화학 장비에서의 용혈을위한 반정량 지표이다. 용혈 지수의 특성이 상업 플랫폼마다 다르기 때문에 용혈 지수의 표준화 또는 일치화는 현재 되어 있지않다. 진단검사의학에서 일치화(harmonization)는 측정 절차와 상관없이 동일한 결과와 동일한 해석을 할 수 있는 능력을 말한다. 임상 환자의 혈액 중에서 무작위로 추출한 40개의 검체를인위적으로 체외 용혈(in vitro hemolysis)시킨다. 혈액 자동화 장비인 XE-2100 (Sysmex, Japan)에서 측정된 혈청 헤모글로빈을 기준으로 검체의 혈청을 0∼300 mg/dL 사이에서 20 mg/dL 차이로 각각을 희석시킨다. 희석된 혈청 헤모글로빈을생화학 자동화 장비 Hitachi-7600 (Hitachi, Japan)으로 측정하여 용혈 지수와 혈청 헤모글로빈을 평가하였다. 용혈 지수와혈청 헤모글로빈은 선형성을 보였으며 용혈 지수 1은 대략 혈청헤모글로빈 20 mg/dL 정도였다. 용혈 거절 기준을 결정하기 위해 용혈 지수를 세 그룹으로 나눈다: 용혈 지수 0∼1, 용혈지수4∼6, 용혈 지수 9∼15. 또 다른 임상 검체 40개의 검체를Hitachi-7600 (Hitachi, Japan)로 각각의 임상 검사 종목을 측정한 후, 각각의 검체를 주사기의 피스톤을 앞·뒤로 움직여 인위적인 체외 용혈을 시키고 Hitachi-7600 (Hitachi, Japan)로측정하였다. 세 그룹 사이의 임상 검사 종목의 용혈로 인한 백분율 차이(percentage difference)를 ANOVA 또는 Kruskal- Wallis test 분석하였다. 사후 검정에서 용혈 지수 0∼1인 그룹과 용혈지수 5∼6인 그룹 사이에 유의한 차이가 나는 종목들은Glucose, creatinine, total protein, AST, direct bilirubin, uric acid, phosphorus, triglyceride, LDH, CPK, magnesium, potassium이였다. 많은 임상 검사 종목들이 용혈지수 4∼6인그룹과 참조 그룹인 용혈지수 0∼1인 그룹 간에 통계적으로 유의미한 차이를 보이기 때문에 용혈 기준은 대략 용혈지수 5 (혈청 헤모글로빈 100 mg/dL)가 적합하다 판단된다.

의료기기 내부 용혈 현상의 예측을 위한 난류 벽 모델의 적용

이승헌(Seunghun Lee),장민욱(Minwook Chang),강성원(Seongwon Kang),허남건(Nahmkeon Hur),김원정(Wonjung Kim) 대한기계학회 2016 대한기계학회 춘추학술대회 Vol.2016 No.12

For medical devices with blood flows, analyzing and reducing hemolysis is a very important topic. The shear stress has been identified as the most important factor for the hemolysis. In turbulent flows, high shear stress values are concentrated near the wall. In case of predicting hemolysis numerically, this leads to a very fine mesh and large computational resource to predict hemolysis accurately. In order to resolve this issue, turbulence wall models are used widely to relax the grid requirements. However, there exists no turbulence wall model for predicting the hemolysis yet. Therefore, the purpose of this study is to develop a turbulence wall model for hemolysis. In order to decrease the numerical error of hemolysis prediction, the blood damage index (BDI) is calculated using different approaches in two divided regions. In a near-wall region, an analytic approach using a modeled velocity profile is used to reduce a numerical error from the sharp velocity gradient in a coarse grid resolution. The Van Driest equation is adopted as the model for the mean velocity profile. In a region far from the wall, a regular numerical discretization is applied. The effectiveness of the proposed wall model is assessed for a few turbulent flows such as the flows inside a channel and cannula. The predicted BDI values present that the proposed wall model shows a significantly improved grid convergence and accuracy compared to a fully numerical approach.

의료 장비 내에서의 용혈에 대한 오일러리안 해석 기법

최종락(Jongrak Choi),허남건(Nahmkeon Hur) 대한기계학회 2020 대한기계학회 춘추학술대회 Vol.2020 No.12

Blood is damaged by hemolysis of red blood cells in transporting the blood driven devices in various medical conditions, leading to blood hemoglobinemia. A hemolysis phenomenon methodology is vital to design in-vitro medical devices, including the cannular, the blood pump, and the oxygenator. In this study, the Eulerian based hemolysis model was developed by comparing the previous experimental results of hemolysis. To cover the short exposure time and high shear stress in the turbulent flow near the blood pumps rotor, a profile of the hemolysis index is adopted by solving the passive scalar transport equation. A good agreement with the modified hemolysis index was obtained in the cannula and FDA blood pump. This method for the hemolysis prediction can be helped the designer of the blood pump with a low computational cost.

용혈이 Vitamin $B_{12}$ / Folic acid 검사 결과에 미치는 영향

김미숙,조윤교,임도화,서소연,Kim, Mi-Sook,Cho, Youn-Kyo,Im, Do-Hw,Seo, So-Yeon 대한핵의학기술학회 2008 핵의학 기술 Vol.12 No.1

배경: Vitamin $B_{12}$ 및 Folic acid는 모두 수용성 비타민의 일종으로 생체 내에서 보조 효소로서의 작용을 가지고 있지만 특히 세포핵의 DNA 합성에 관여하고 있다. 주로 거대 적아구성 빈혈의 감별 진단과 임신 중 엽산의 저장을 평가하기 위해 측정한다. 용혈 혈청은 혈구 중의 엽산과 비타민 $B_{12}$가 용출하기 때문에 사용할 수 없다고 알려져 있으나 수탁 검사기관인 본원의 특성상 이 내용을 충실히 따르기에는 어려운 경우가 있다. 이에 용혈의 정도가 검사 결과치에 어느 정도 영향을 미치는가에 대해 알아보고자 검사를 시행해 보았다. 방법: 용혈되지 않은 검체를 대조군으로 하고 기계적으로 용혈을 일으켜 용혈 정도에 따라 A, B, C그룹으로 분리하여 용혈 검체를 그룹별 각각 10개씩 준비한다. 경쟁반응 원리인 M사의 Vitamin $B_{12}$ [$^{57}Co$]/Folate [$^{125}I$] 시약으로 검사를 실시한다. 결과: Vitamin $B_{12}$는 검체의 용혈이 결과 값에 큰 영향을 미치지 않았다. 하지만 Folic acid의 경우에는 용혈의 정도에 따라 값이 증가 하였으며, 심하게 용혈 된 경우 모두 표준곡선의 최고치의 값(20 ng/ml 이상)을 확인할 수 있었다. 고찰: 수탁기관인 본원에서는 Vitamin $B_{12}$ 및 Folic acid 검사 의뢰 병원에 검체의 용혈이 검사 결과에 미치는 영향과 검체의 용혈을 막기 위해 채혈 시 주의할 점들을 홍보하고, 분리, 보관 시 세심한 주의를 기울여야 할 것이다. 위의 결과에서 보았듯이 용혈된 검체로 Folic acid검사를 해서는 안 되며 Vitamin $B_{12}$ 또한 결과 값에는 큰 영향을 미치지는 않지만 용혈에 주의할 필요가 있다. Purpose: Vitamin $B_{12}$ and folic acid are type of water-soluble vitamin and those work as a secondary vital enzyme, but especially those are involved in the nuclear DNA synthesis. Those are mainly measured in order to diagnose megaloblastic anemia and to assess the storage of folic acid during pregnancy. It is generally known that the hemolyzed serum is useless for folic acid and vitamin $B_{12}$, but it is not easy to abide by this information because our lab (Samkwang Medical Laboratories) is reference laboratory. We tested how much the extent of hemolyzed serum had influence on the results. Materials and Method: We performed the tests of vitamin $B_{12}$, folic acid. For the hemolysis effect study, we used 40 cases. According to the level of A, B and C groups, each group had 10 samples which were mechanically hemolyzed serum. Results: Hemolysis did not affect the vitamin $B_{12}$ results. However in case of folic acid, the value increased according to the degree of hemolysis. And severe hemolyzed cases had the highest value (greater than 20 ng/ml). Conclusions: Preventing the hemolysis, it has to be informed that blood-collecting, separation and storage are performed more carefully. As you see from the above results, hemolyzed serum is not proper for folic acid test, and hemolysis does not affect the results of vitamin $B_{12}$. However, for the more accurate results, it is necessary to avoid hemolysis.

Scoring System for Detecting Spurious Hemolysis in Anticoagulated Blood Specimens

유길성,김주원,어영,윤갑준,박순덕,윤광로 대한진단검사의학회 2015 Annals of Laboratory Medicine Vol.35 No.3

Background: The identification of in vitro hemolysis (IVH) using a hematology analyzer is challenging because centrifugation of the specimens cannot be performed for cell counts. In the present study, we aimed to develop a scoring system to help identify the presence of hemolysis in anticoagulated blood specimens. Methods: Thirty-seven potassium EDTA anticoagulated blood specimens were obtained, and each specimen was divided into 3 aliquots (A, B, and C). Aliquots B and C were mechanically hemolyzed by aspirating 2 and 5 times, respectively, using a 27-gauge needle and then tested; aliquot A was analyzed immediately without any hemolysis. After the cells were counted, aliquots B and C were centrifuged and the supernatants were tested for the hemolytic index and lactate dehydrogenase levels. Results: The 4 hematologic parameters were selected and scored from 0 to 3 as follows:<34.0, 34.0-36.2, 36.3-38.4, and ≥38.5 for mean cell hemoglobin concentration (MCHC, g/dL); <0.02, 0.02, 0.03, and ≥0.04 for red blood cell ghosts (1012/L); <0.13, 0.13-0.38, 0.39-1.30, and ≥1.31 for difference value (g/dL) of measured hemoglobin and calculated hemoglobin; and <0.26, 0.26-0.95, 0.96-3.34, and ≥3.35 for difference value (g/dL) of MCHC and cell hemoglobin concentration mean. The hemolysis score was calculated by adding all the scores from the 4 parameters. At the cutoff hemolysis score of 3, the IVH of aliquots B and C were detected as 64.9% and 91.9%, respectively. Conclusions: The scoring system might provide effective screening for detecting spurious IVH.

무기수은제에 의한 적혈구 용혈의 특성

송은주,정영미,조종후 한국수의공중보건학회 2002 예방수의학회지 Vol.26 No.3

This study was examined to find out the possibility of membrane lipid peroxidation as the cause on mercuric chloride-induced hemolysis of indigenous Korean goat erythrocytes. In 0.03% erythrocyte suspension treated with 2 10 M of HgCl₂, hemolysis tended to occur HgCl₂-dependently. Temporary resistance on HgCl₂-induced hemolysis was shown in all groups 15 minutes later which is different from that on H₂O₂ treated hemolysis. Changes of thiobarbituric acid(TBA) value and fluorescence intensity were measured to investigate the possibility of lipid peroxidation in 2 20% hemolytic groups of HgCl₂-treated 4% erythrocyte suspension. There were not only no changes of TBA value rated as lipid peroxidation by H₂O₂, but also even decrement of fluorescence intensity. These results suggest that HgCl₂^(-) induced hemolysis is not related to membrane lipid peroxidation.