• Title/Summary/Keyword: Experimental Kit

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Charaeteristics and Applications of the Established Testosterone Immunoassays (Testosterone 면역측정법의 정립 특성과 응용)

  • Lee, Chang-Joo;Lee, Joon-Yeoung;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.5 no.1
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    • pp.59-71
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    • 2001
  • Since the first radioimmunoassay (RIA) was developed in 1970s, many conventional RIAs and non-isotopic immunoassays (NIA) had been developed in which the degree of precision, accuracy, specificity and practicability have progressively increased. Recently ultrasensitive assay method at femtogram to determine testosterone in serum, saliva and feces is required for the study of sexual dysfunctions in male and female, monitoring the psychological stress and conditions, aging process such as menopause and partial androgen deficiency in aging male, the hormonal changes of small experimental animals etc. This review discussed the recentd evelopments of steroid assay methods, based upon the testosterone assay results of authors far 20 years, and the problems associated the assay set-up, the characterizations and applications of the established procedures, and desifls of assay, reliablity criteria, and the practical aspects of assay set-up and application, based upon the data of the authors. The present study demonstrates the general problems methods to be consider in order to set up the highly sensitive assay methods and to increase the assay quality and the necessity of assay quality control program. To improve the assay quality of each laboratory and to compare the assay results in homeland, the national QC programs should be organized.

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A Study on the Variations of the Body Trunk Temperature by the Drug-Pad Moxibustion Method (약패드 뜸 방식을 이용한 체간온도변화에 대한 연구)

  • Yoon Dong-Eop;Jo Bong-Kwan
    • The Transactions of the Korean Institute of Electrical Engineers D
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    • v.55 no.8
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    • pp.386-396
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    • 2006
  • We implemented the Drug-Pad Moxibustion Method in order to improve the conventional moxibustion therapy. This method is aimed to eliminate burning wounds and smoke, which are the defects of conventional moxibustion therapy. And we performed to verify the efficiency by comparing the Drug-Pad Moxibustion Method with the conventional Indirect Moxibustion Therapy. We measured the body heat and the lasting time of blood circulation improvement using thermography. The moxibustion therapy has two kinds of effects: The formers are pharmacological effects of the Moxa's vasodilators and antioxidants. The latters are thermal effects which cause improvement of the blood circulation. To remove the demerits without omission of above therapeutic effects, we extracted the vasodilators and antioxidant compounds from the Moxa-$CH_2Cl_2$ fraction Moxa-EtOAc and composed the moxibustion kit with $(Ba_{0.8}\;Sr_{0.2})_{0.996}\;Y_{0.004}\;TiO_2+0.5_{WT}\;SiO_2%$ Positive Temperature Coefficients Thermistor. The experimental demonstrations have been made by the stimulating the spot which is CV4(Kwan-Won), CV8(Shin-Guel), CV12(Jung-Wan) acupuncture points of the conception vessel meridian(CV). And stimulating time was one hour. We divided the subjects into 5 groups such as no stimulation group, conventional Indirect Moxibustion group, only Drug-Pad stimulation group, only heat stimulation group, and Drug-Pad Moxibustion group. In the different cases, we have measured the body heat in pre-stimulation, just after stimulation, 2 hours after, and 4 hours after. The body heats of the group who were stimulated by the Drug-Pad Moxibustion Method were increased by over the $2^{\circ}C$. And the body heats of the group who were stimulated by the Indirect Moxibustion Method were increased by average the $1^{\circ}C$. We have evaluated that the Drug-Pad Moxibustion Method is improvement on the conventional Indirect Moxibustion Method by the heat-increasing rate is 200% and the lasting time is 150% with the body heat of the abdominal region. In the conclusions, We have implemented the Drug-Pad Moxibustion Method and evaluated the efficiency of the Drug-Pad Moxibustion Method comparing with the conventional Indirect Moxibustion Method.

Effects of a Plant Water Extract Mixture(Ixeris sonchifolia Hance, Oenanthe javanica, Fagopyrum esculentum Moench, Hizikia fusiforme, Zingiber officinale Roscoe) on Mouse Immune Cell Activation (식물 혼합(고들빼기, 돌미나리, 메밀, 톳, 생강) 추출물이 마우스 면역 세포 활성에 미치는 영향)

  • Kim, Jung-Hee;Kim, Hyun-Sook;Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.20 no.1
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    • pp.74-78
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    • 2007
  • Ixeris sonchifolia Hance(Godulbaegi), Oenanthe javanica(Dolminari), Fagopyrum esculentum Moench(Buckwheat), Hizikia fusiforme(Seaweed Fusiforme) and Zingiber officinale Roscoe(Ginger) have all been used as one of the traditional remedies as well as food source. There are few studies However, on their immunomodulating effects have been reported. We previously reported that ex vivo supplementation of each of the Ish, Oj, Fem, Hf and Zor water extracts enhanced the splenocytes proliferation compared to the control group. In this study, the combined immunomodulative effects of a plant water extract mixture containing these five food sources(Ish+Oj+Fem+Hf+Zor) was compared to the individual effect of each. The production of cytokine(IL-1${\beta}$, IL-6, and TNF-${\alpha}$), secreted by macrophages stimulated with LPS or without, were detected via ELISA assay using a cytokine kit. After 48hrs of incubation with mitogen(ConA or LPS) stimulation, the mouse splenocyte proliferation in the experimental group had significantly increased at two different concentrations compared to the control group. The results of this study may suggest that the supplementing with a plant water extract mixture could regulate immune function by increasing splenocyte proliferation as well as enhance immune function by regulating the cytokine production capacity activated macrophages in mice.

Application of Cumulus Cells as Factors to Predict the Outcome of IVF-ET (체외수정시술의 결과를 예측할 수 있는 인자로서 난구세포의 활용에 관한 연구)

  • Kim, Kwang-Dae;Kim, Ki-Hyung;Na, Yong-Jin;Lee, Kyu-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.3
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    • pp.419-432
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    • 1999
  • Objective: To establish the evaluation system of the quality of oocytes on the basis of the incidence of cumulus cells apoptosis, to investigate the relationships beween the incidence of cumulus cells and the outcomes of IVF-ET. Method: Thirth-four cycles undergoing controlled ovarian hyperstimulation for IVF-ET with tubal infertility (23 cycles) or unexplained infertility (11 cycles) were included in this study. Cumulus cell masses surrounding mature oocyte and co-culture of embryos with autologous cumulus cells during IVF-ET process. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. The effect of co-culture using cumulus cells and the incidence of cumulus cells apoptosis. Results: The results were as follows: 1. The incidence of apoptosis in cumulus cells markedly increased in patients aged 40 or over, while the fertilization rate was greatly decreased in those age group. 2. Apoptosis in cumulus cells was found in both the fertilized oocytes and unfertilized oocytes, but the incidence of apoptosis was higher in unfertilized oocytes. 3. There is no clear correlation between apoptosis in cumulus cells and the number of oocytes retrieved. However, the incidence of apoptosis was increased when the number of oocytes retrieved was 5 and fewer in comparison with $6{\sim}10$. 4. Embryo grade was significantly affected by the incidence of apoptosis in cumulus cells. 5. Pregnancy rate of IVF-ET per cycle was 29.4%, and the pregnant group had the higher fertilization rate and a significantly lower incidence of apoptosis in cumulus cells compared with the nonpregnant group. 6. When cumulus cells were used as helper cells in the co-culture of the embryo, in vitro activity of cumulus cells based on morphological change and proliferation did not influence the quality of embryo, but was closely associated with the implantation rate and pregnancy rate, which was enhanced when morphological changes and proliferation of cumulus cells was more active. 7. This difference in the outcome of IVF-ET according to in vitro activity of cumulus cells used for co-cultue was not associated with the incidence of apoptosis in cumulus cells; but rather had likely relations with the different secretion pattern of protein, which may be an embryo trophic factor by cumulus cells. Conclusion: These results suggest that the incidence of apoptosis in cumulus cells can be used in predicting oocyte qualities and the outcomes of IVF-ET. And the effect of co-culture largely depends on the in vitro activity of cumulus cells as well.

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Nitric oxide, 8-hydroxydeoxyguanosine, and total antioxidant capacity in human seminal plasma of infertile men and their relationship with sperm parameters

  • Gholinezhad, Maryam;Aliarab, Azadeh;Abbaszadeh-Goudarzi, Ghasem;Yousefnia-Pasha, Yousefreza;Samadaian, Niusha;Rasolpour-Roshan, Korush;Aghagolzadeh-Haji, Hemat;Mohammadoo-Khorasani, Milad
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.1
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    • pp.54-60
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    • 2020
  • Objective: Oxidative stress plays a key role in the pathogenesis of male infertility. But, the adverse effects of oxidative biomarkers on sperm quality remain unclear. This study aimed to investigate the levels of nitric oxide (NO), 8-hydroxydesoxyguanosine (8-OHdG), and total antioxidant capacity (TAC) oxidative biomarkers in seminal plasma and their relationship with sperm parameters. Methods: A total of 77 volunteers participated in the study, including fertile (n = 40) and infertile men (n = 37). NO, 8-OHdG, and TAC levels were measured using the ferric reducing ability of plasma, Griess reagent method and an enzyme-linked immunosorbent assay kit, respectively. Results: The mean values of sperm parameters in the infertile group were significantly lower than those in the fertile group (p< 0.001). The mean 8-OHdG in the seminal plasma of infertile men was significantly higher (p= 0.013) than those of controls, while the mean TAC was significantly lower (p= 0.046). There was no significant difference in NO level between the two groups. The elevated seminal 8-OHdG levels were negatively correlated with semen volume, total sperm counts and morphology (p< 0.001, p= 0.001 and p= 0.052, respectively). NO levels were negatively correlated with semen volume, total sperm counts and morphology (p= 0.014, p= 0.020 and p= 0.060, respectively). Positive correlations between TAC and both sperm count and morphology (p= 0.043 and p= 0.025, respectively) were also found. Conclusion: These results suggested that increased levels of NO and 8-OHdG in seminal plasma could have a negative effect on sperm function by inducing damage to the sperm DNA hence their fertility potentials. Therefore, these biomarkers can be useful in the diagnosis and treatment of male infertility.

An Empirical Study of the Recovery Experiment in Clinical Chemistry (임상화학검사실에서 회수율 실험의 실증적 연구)

  • Chang, Sang-Wu;Lee, Sang-Gon;Song, Eun-Young;Park, Yong-Won;Park, Byong-Ok
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.184-188
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    • 2006
  • The purpose of the recovery experiment in clinical chemistry is performed to estimate proportional systematic error. We must know all measurements have some error margin in measuring analytical performance. Proportional systematic error is the type of error whose magnitude increases as the concentration of analyte increases. This error is often caused by a substance in the sample matrix that reacts with the sought for analyte and therefore competes with the analytical reagent. Recovery experiments, therefore, are used rather selectively and do not have a high priority when another analytical method is available for comparison purposes. They may still be useful to help understand the nature of any bias revealed in the comparison of kit experiments. Recovery should be expressed as a percentage because the experimental objective is to estimate proportional systematic error, which is a percentage type of error. Good recovery is 100.0%. The difference between 100 and the observed recovery(in percent) is the proportional systematic error. We calculated the amount of analyte added by multiplying the concentration of the analyte added solution by the dilution factor(mL standard)/(mL standard + mL specimen) and took the difference between the sample with addition and the sample with dilution. When making judgments on method performance, the observed that the errors should be compared to the defined allowable error. The average recovery needs to be converted to proportional error(100%/Recovery) and then compared to an analytical quality requirement expressed in percent. The results of recovery experiments were total protein(101.4%), albumin(97.4%), total bilirubin(104%), alkaline phosphatase(89.1%), aspartate aminotransferase(102.8), alanine aminotransferase(103.2), gamma glutamyl transpeptidase(97.6%), creatine kinase(105.4%), lactate dehydrogenase(95.9%), creatinine(103.1%), blood urea nitrogen(102.9%), uric acid(106.4%), total cholesterol(108.5), triglycerides(89.6%), glucose(93%), amylase(109.8), calcium(102.8), inorganic phosphorus(106.3%). We then compared the observed error to the amount of error allowable for the test. There were no items beyond the CLIA criterion for acceptable performance.

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Effect of Fish Meal Replacement on Insulin-like Growth Factor-I Expression in the Liver and Muscle and Implications for the Growth of Olive Flounder Paralichthys olivaceus (사료의 어분함량대체가 넙치(Paralichthys olivaceus)의 간과 근육 내 인슐린유사성장인자의 발현과 체성장에 미치는 영향)

  • Park, Su-Jin;Moon, Ji-Sung;Seo, Jin-Song;Nam, Taek-Jeong;Lee, Kyeong-Jun;Lim, Sang-Gu;Kim, Kang-Woong;Lee, Bong-Joo;Hur, Sang-Woo;Choi, Youn Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.2
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    • pp.141-148
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    • 2019
  • This study examined the effect of insulin-like growth factor (IGF)-I expression in the liver and muscle on the growth of Paralichthys olivaceus fed diets low in fish meal. A feeding experiment was conducted at Jeju National University, Jeju Island, Korea. Groups of P. olivaceus (total initial weight: 200 g) were maintained for 20 weeks on one of five experimental diets containing different proportions of fish meal. Diets containing 0%, 20%, 30%, 40%, and 50% fish meal were labeled FM0, FM20, FM30, FM40, and FM50, respectively. Fish growth was observed every 4 weeks during the feeding experiment, and plasma and liver and muscle tissues were sampled. Plasma IGF-I levels were analyzed using an ELISA kit. The mechanism of IGF-I receptor signaling was examined using immunoblotting and reverse transcription-polymerase chain reaction. The greatest total weight increase was observed in the FM30 group. In parallel, plasma levels of IGF-I and IGF-binding protein were highest in the FM30 group, and mRNA and protein expression were also significantly higher in this group. The first step in the IGF-I signaling pathway, tyrosine-phosphorylation checking, occurred smoothly until 20 weeks. These results suggest that a dietary ratio of 30% fish meal best promotes growth in this species. The IGF-I signaling pathway in the liver and muscle is associated with growth in P. olivaceus.

The Effect of Ampelopsis japonica (Thunb.) Makino on Osteoclastogenesis and Expression of Osteoclast-Related Gene (백렴(白蘞)의 파골세포 분화 및 관련 유전자 발현 억제에 미치는 영향)

  • Hongsik Kim;Sumin Lee;Minsun Kim;Jae-Hyun Kim;Yejin Kang;Seoung Jun Kwon;Youngwoo Nam;Seungwoo Yoo;Hong-Seok Choi;SeonJin Huh;Youngjoo Sohn;Hyuk-Sang Jung
    • The Korea Journal of Herbology
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    • v.38 no.5
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    • pp.9-19
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    • 2023
  • Objectives : Osteoporosis is a systemic skeletal disorder characterized by reduced bone mineral density and increased risk of fractures. Bisphosphonates and selective estrogen receptors, which are bone resorption inhibitors that are currently widely used as osteoporosis treatments, show serious side effects when administered for a long time. Research on bone resorption inhibitors that complement the problems of existing treatments is needed. The purpose of this study was to investigate the effect of inhibiting osteoclast differentiation and activity on the tuberous root of Ampelopsis japonica (Thunb.) Makino (AM). Methods : After extracting AM using distilled water and ethanol, the inhibitory effects of the two solvents on osteoclast differentiation were compared using the RANKL-induced in vitro experimental model and the TRAP assay kit. The impact of AM on bone resorption was investigated through the pit formation assay, and its effect on F-actin formation was assessed through fluorescent staining. Additionally, protein and mRNA expression levels of osteoclast differentiation markers (NFATc1, c-Fos, TRAP and ATP6v0d2) and resorption markers (MMP-9, CTK, and CA2) were analyzed via western blot and RT-PCR. Results : AM treatment significantly decreased the number of TRAP-positive cells and pit formation area. Furthermore, AM suppressed both the protein and mRNA expression of NFATc1 and c-Fos, key transcription factors involved in osteoclast differentiation and it downregulated the expression of osteoclast-associated genes such as TRAP, CTK, MMP-9, CA2, and ATP6v0d2. Conclusions : These results suggest that AM can inhibit bone resorption and osteoclast differentiation, indicating its potential for use in the treatment and prevention of osteoporosis.

The Effect of the IGF-I treated Gingival and Periodontal Ligament Fibroblast on Osteoblasts (IGF-I으로 처리한 치은 및 치주인대 섬유모세포가 골모세포에 미치는 영향)

  • Kim, Mi-Jeong;Yang, Won-Sik
    • The korean journal of orthodontics
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    • v.31 no.6 s.89
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    • pp.589-600
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    • 2001
  • Insulin-like growth factor I (IGF-I) has the local tissue regulating actions. In bone, IGF-I increases the replication of osteoblastic lineage, probably preosteoblasts, and enhances osteoblastic collagen synthesis and matrix composition rates. The purpose of this study was to investigate the local regulatory effect of IGF-I on periodontium totally, both in an autocrine and paracrine manner. To examine the effect of IGF-I directly on osteoblast (OB) of test rats, and indirectlv on OB via periodontal ligament fibroblast (PDLF), and the effect of gingival fibroblast (GF) on OB via cellular paracrine manner for the understanding of humoral action of adjacent tissue, GF and PDLF were obtained from male Sprague-Dawley rats of six to eight weeks of age. OB was obtained iron frontal and parietal calvarial bone of Sprague-Dawley 21day-old-fetus. After each tell was Incubated 24 hours, for collecting conditioned medium, different concentrations of IGF-I (1,10,100 ng/ml,1ml/well) was adding in the GF, PDLF cells, and the supernatant from these cultures was put into the primary OB culture with $1{\times}10^4$cell/ml/well. The experimental group was divided into six groups control OB, IGF-I treated OB, OB culture with conditioned medium from PDLF, OB culture with conditioned medium from IGF-I treated PDLF, OB culture with conditioned medium from GF, OB culture with conditioned medium from IGF-I treated GF. After final IGF-I treatment, OB was Incubated for 24 hours, and alkaline phosphatase activity assay, BMP expression, cell proliferation measurement using MTT assay, total protein measurement, Collagen synthesis assay using western blot, and examination of bone nodule synthesis were done. Alkaline phosphatase expressions were increased in the group of PDLF-IGF-I supernatant treatment. Direct IGF-I treatment with concentrations of 100ng/m1 showed increased viable tell number measured by MTT assay. And IGF-I treatment did not increase total protein amount. The entire experimental group showed BMP2, 4 expression in western blot, and there was no significant difference between control and experimental groups. These results suggested that supernatant from PDLF effects on increasing cellular activities of OB regardless of IGF-I, and at high concentration, IGF-I increases OB tell proliferation.

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Application of Westgard Multi-Rules for Improving Nuclear Medicine Blood Test Quality Control (핵의학 검체검사 정도관리의 개선을 위한 Westgard Multi-Rules의 적용)

  • Jung, Heung-Soo;Bae, Jin-Soo;Shin, Yong-Hwan;Kim, Ji-Young;Seok, Jae-Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.16 no.1
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    • pp.115-118
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    • 2012
  • Purpose: The Levey-Jennings chart controlled measurement values that deviated from the tolerance value (mean ${\pm}2SD$ or ${\pm}3SD$). On the other hand, the upgraded Westgard Multi-Rules are actively recommended as a more efficient, specialized form of hospital certification in relation to Internal Quality Control. To apply Westgard Multi-Rules in quality control, credible quality control substance and target value are required. However, as physical examinations commonly use quality control substances provided within the test kit, there are many difficulties presented in the calculation of target value in relation to frequent changes in concentration value and insufficient credibility of quality control substance. This study attempts to improve the professionalism and credibility of quality control by applying Westgard Multi-Rules and calculating credible target value by using a commercialized quality control substance. Materials and Methods : This study used Immunoassay Plus Control Level 1, 2, 3 of Company B as the quality control substance of Total T3, which is the thyroid test implemented at the relevant hospital. Target value was established as the mean value of 295 cases collected for 1 month, excluding values that deviated from ${\pm}2SD$. The hospital quality control calculation program was used to enter target value. 12s, 22s, 13s, 2 of 32s, R4s, 41s, $10\bar{x}$, 7T of Westgard Multi-Rules were applied in the Total T3 experiment, which was conducted 194 times for 20 days in August. Based on the applied rules, this study classified data into random error and systemic error for analysis. Results: Quality control substances 1, 2, and 3 were each established as 84.2 ng/$dl$, 156.7 ng/$dl$, 242.4 ng/$dl$ for target values of Total T3, with the standard deviation established as 11.22 ng/$dl$, 14.52 ng/$dl$, 14.52 ng/$dl$ respectively. According to error type analysis achieved after applying Westgard Multi-Rules based on established target values, the following results were obtained for Random error, 12s was analyzed 48 times, 13s was analyzed 13 times, R4s was analyzed 6 times, for Systemic error, 22s was analyzed 10 times, 41s was analyzed 11 times, 2 of 32s was analyzed 17 times, $10\bar{x}$ was analyzed 10 times, and 7T was not applied. For uncontrollable Random error types, the entire experimental process was rechecked and greater emphasis was placed on re-testing. For controllable Systemic error types, this study searched the cause of error, recorded the relevant cause in the action form and reported the information to the Internal Quality Control committee if necessary. Conclusions : This study applied Westgard Multi-Rules by using commercialized substance as quality control substance and establishing target values. In result, precise analysis of Random error and Systemic error was achieved through the analysis of 12s, 22s, 13s, 2 of 32s, R4s, 41s, $10\bar{x}$, 7T rules. Furthermore, ideal quality control was achieved through analysis conducted on all data presented within the range of ${\pm}3SD$. In this regard, it can be said that the quality control method formed based on the systematic application of Westgard Multi-Rules is more effective than the Levey-Jennings chart and can maximize error detection.

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