• Title/Summary/Keyword: RIA

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Physiological Regulation of Luteinizing Hormone(LH) Expression in Rat Mammary Gland during Differentiation (분화중인 흰쥐 유선내 Luteinizing Hormone (LH) 유전자 발현의 생리적인 조절)

  • 이성호
    • Development and Reproduction
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    • v.5 no.2
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    • pp.175-180
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    • 2001
  • The ectopic expression of gonadotropin releasing hormone(GnRH and luteinizing hormone(LH) in several tissues is a quite intriguing phenomenon. Recently, the presence of GnRH and its receptor has been clearly demonstrated in rodents and human mammary gland. In this context, one can postulate that the presence of local circuit composed of GnRH and LH in the gland. The present study was undertaken to elucidate whether there is a correlation between the LH expression in rat mammary gland and physiological status during the process of mammary differentiation. LH contents in mammary gland from cycling to weaning rats were measured by radioimmunoassay(RIA). In cycling rats, changes of the LH level in both serum and mammary gland showed similar pattern as the highest level in proestrus and the lowest level in diestrus II stage. While the serum LH levels were fluctuated from pregnant through involution stage, a sharp decline of mammary LH contents was observed in the lactating rats. This decrement was recovered in involuting rats to the level of proestrus stage. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analyses demonstrated that the transcriptional activities of the mammary LH and GnRH were increased from diestrus I stage to estrus stage, and the increased levels were maintained in pregnant, lactation and involution stages. To test the hypothesis that the alteration in mammary LH expression might be steroid-dependant, ovariectomy(OVX) and steroid supplement model was employed. As expected, supplement of estradiol(E$_2$) after OVX remarkably decreased serum LH level compared to that in serum from vehicle-only treated rats. Likewise, administration of E$_2$ significantly reduced the mammary LH content. The present study demonstrated that (i) the LH expression in mammary gland could be altered by some physiological parameters such as estrous cycle, pregnancy, lactation and involution, and (ii) ovarian steroid especially estrogen seems to be one of major endocrine factors which are responsible for regulation of mammary LH expression.

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Effect of Di(2-ethyl hexyl)phthalate(DEHP) on the Onset of Puberty in Female Rat (암컷 흰쥐의 사춘기 개시에 미치는 di(2-ethyl hexyl)phthalate(DEHP)의 효과)

  • Lee, Kyeung-Yeup;Lee, Sung-Ho
    • Development and Reproduction
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    • v.10 no.2
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    • pp.147-154
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    • 2006
  • Phthalates such as di(2-ethyl hexyl)phthalate(DEHP) are industrial chemicals with wide-ranging human exposures because of their use in plastics and other common consumer products. Consequently, their adverse effects as endocrine disruptor in the reproductive physiology of both laboratory rodents and human have been studied extensively. The present study was undertaken to examine whether prepubertal exposure to DEHP affects on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. DEHP(100mg/kg/day) was administered daily from postnatal day 25(PND 25) through the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the next day. Gross anatomy and weight of reproductive tissues were compared to test the DEHP's effects on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in the tissues. Specific radioimmunoassay was carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptor(PR), total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a result, delayed VO was shown in the DEHP group(PND $37.3{\pm}0.7$) compared to the control group(PND $35.3{\pm}0.7$; p<0.05). DEHP treatment significantly decreased the wet weight of ovaries and uteri compared to the control group(p<0.05). Interestingly, elevation of serum LH levels was shown in the DEHP group(p<0.05). Graafian follicles and corpora lutea were observed only in the ovaries from the control animals. Numerous primary, secondary follicles and small atretic follicles were observed in the ovaries from DEHP-treated animals. Similarly, hypotrophy of luminal and glandular uterine epithelium was found in the DEHP-treated group. These effects were probably due to the inhibitory effects of DEHP on the synthesis and secretion of estrogen from granulosa cells. In the semiquantitative RT-PCR studies, the transcriptional activities of PR in both ovary(p<0.05) and uterus(p<0.01) from DEHP-treated animals were significantly lower than those from the control animals. The present studies demonstrated that the acute exposure to DEHP during the critical period of prepubertal stage could inactivate the reproductive system resulting delayed puberty in female rats.

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Evaluation of Usefulness for Anti-TPO Antibody Test in Item of the Medical Examination (건강검진 종목으로서 항갑상선 과산화효소 항체검사의 유용성에 대한 평가)

  • Kim, Yun-Hyun;Shin, Yong-Hwan;Kim, Ji-Young;Seok, Jae-Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.13 no.1
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    • pp.112-115
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    • 2009
  • Purpose: The test had been applied for outpatient by end of 2006, however, it has been included in the medical examination since January 2007, as demand and interests have been gradually increasing in the thyroid gland disease and cancer. thus, we would necessarily evaluate usefulness of the test by comparing the number of patients who are diagnosed as "benignancy" by the medical test with the number of outpatient who attend and are diagnosed as autoimmune thyroid disease among the benign patient, in samsung medical center for a certain period. Materials and Methods: Based on the result for Anti-TPO Antibody test by RIA for the 12,937 patients in samsung medical center from October 2007 to March 2008, for six months, benignancy rate classified by sex and age is measured statistically and number of the patients who are diagnosed as autoimmune thyroid disease are kept tracked on. Results: According to the analysis of the Anti-TPO antibody test 1,135 of 12,937, which is 8.77% are benign and 218 treated patient of them, which is 19.2%, were diagnosed as autoimmune thyroid disease. Conclusions: Based on the statistics, usefulness of the test seem to have co relationship with derivation of autoimmune thyroid disease. this is 19.2% of probability relatively high. this figure, however, does not have strong relationship with specialty of the disease. Thus screening test seems to have somewhat effectiveness, considering other experiments and their margin.

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The Study on the Dilution Time of Radioactive Tracer in Estradiol Measurement (Estradiol 검사 시 방사성 추적자의 희석시간에 대한 고찰)

  • Lee, Hae Yeon;Seo, Han Kyung;Jang, Yi Sun;Kim, Hee Jeoung
    • The Korean Journal of Nuclear Medicine Technology
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    • v.21 no.2
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    • pp.44-48
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    • 2017
  • Purpose Estradiol (E2) is a steroid hormone mainly produced in women and is a useful indicator for diagnosis of gynecological diseases, menstrual cycle, menopause, and precocious puberty. E2 measurement is performed by diluting the $^{125}I$ radioactive tracer and tracer buffer in the kit. However, It was not precisely specified when the period of tracer is available after activating. The purpose of this study was to determine the appropriate dilution time based on the measurement value with dilution time. Materials and Methods From December 2016 to February 2017, 60 E2 samples with concentrations ranging from 8 to 4577 pg/mL were divided into low, medium, and high concentrations. Dilution of the $^{125}I$ tracer was performed on a 230 RPM agitator for 30 minutes, 1 hour 30 minutes, and 2 hours 30 minutes, respectively. 24 hour dilution was gently shaken and refrigerated. To verify the difference and significance of the results according to the dilution time, a test of normality was performed using SPSS 18.0 and analyzed by Kruskal-Wallis test. The measured value according to the dilution time was compared with the interquartile range of the absolute error. Results The results of Kruskal-Wallis test were not significant (P>0.05). Measurement results are showed as interquartile range of absolute error. At low concentration, it is 0.052 between 1 hour 30 minutes and 2 hours 30 minutes, and 0.105 between 30 minutes and 1 hour 30 minutes. At medium concentration, 0.062 between 30 minutes and 1 hour 30 minutes, and 0.038 between 1 hour 30 minutes and 2 hours 30 minutes. At high concentration, it is 0.029 between 1 hour 30 minutes and 2 hours 30 minutes, and 0.06 between 2 hours 30 minutes and 24 hours. Conclusion There were no statistically significant differences. However, the change in the measured value is the smallest between 1 hour and 30 minutes to 2 hours and 30 minutes. Therefore, we recommend diluting time between 1 hour 30 minutes and 2 hours 30 minutes.

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Analysis of Immunogenicity after Hepatitis B Vaccination in Korea by Literature Review (문헌 분석에 의한 B형 간염 백신의 항체 양전률의 비교)

  • Roh, Hye Ok;Lee, Woo Gill;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.5 no.2
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    • pp.245-257
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    • 1998
  • Purpose : Although hepatitis B vaccine has been available to general population in Korea since 1983, it was difficult to compare various types of hepatitis B virus(HBV) vaccines primarily due to the differences in vaccination schedule, dosage, test methods and seropositive antibody level. In this study we reviewed the results of previous studies published in Korea, which include antibody positive rates and antibody titers of various vaccines, and examined the immunogenicity of these HBV vaccines. Methods : Studies published in medical journals, university journals concerning antibody positive rates following hepatitis B vaccination were reviewed. Inclusion criteria were those studies in which seroprotective antibody rate of 10mIU/mL or the sample ratio unit of 10 RU were used as the cut-off value and in which the test methods were RIA or ELISA. Exclusion criteria were; 1) unclear or inconsistent vaccine dosage, 2) no record of antibody titers or seroconversion rate, 3) no defined antibody rate or ratio for positive rating and 4) the vaccination schedule other than 0-1-2 months or 0-1-6 months. Results : 23 out of 52 studies were subjected for the review for seroconversion rates. 1) As for the immunogenicity in each age group, the seroconversion rates of Hepaccine(Cheil Jedang) were 85.1% in infants, 83.3% in children and 62.7% in adults, indicating higher rates in infants and children compared to adults(P<0.01). The seroconversion rates of Hepavax(Korea Green Cross) were 84.7%, 81.1% and 90.8%, indicating higher rates in infants and adults compared to children(P<0.01). 2) The seroconversion rate of Hepavax was 85.6% with 0-1-6 mo. schedule, 78.5% with 0-1-2 mo. schedule with a statistically significant difference(P<0.01). 4) There was no difference of seroconversion rates between the two doses of Hepavax, $5{\mu}g$ and $10{\mu}g$ in infants and children. 5) In adults the seroconversion rates were 62.7% with Hepaccine, 90.8% with Hepavax, and 94.8% with Engerix-B(SmithKline Beecham). Conclusion : In Korea, the incidence of chronic hepatitis B is high and changing the schedule in vaccination cannot contribute to the increase of the serocoversion rate. And in order to maximize immunogenicity, more effective vaccines as well as more proper vaccination methods should be used.

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Hepatitis B Virus Infection Rate of Medical School Students in Taegu (대구시내(大邱市內) 의과대학생(醫科大學生)들의 B형(型)바이러스성(性) 간염감염율(肝炎感染率))

  • Park, Jung-Han;Youn, Tae-Hyun;Chun, Byung-Yeol;Song, Jung-Hup
    • Journal of Preventive Medicine and Public Health
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    • v.20 no.1 s.21
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    • pp.129-136
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    • 1987
  • To determine the hepatitis 8 virus infection rate of the medical school students and appropriate time for immunization with hepatitis B vaccine,355 students in the 1st, 2nd and 3rd grades of Medical School of Kyungpook National University who had not been vaccinated and volunteered to participate in this study were tested for HBsAg, anti-HBs and anti-HBc with radioimmunoassay method (Abbott Lab. kit). A questionnaire was administered to ask the history of transfusion, acupuncture and surgery. HBsAg positive students were retested 16 months after the initial test. Overall HBsAg positive rate was 6.8% and the age adjusted rate for male (7.2%) was higher than that for female (4.9%). Anti-HBs positive rate was 35.3% (36.1% for male, 37.9% for female) and anti-HBc positive rate was 45.5% (46.5% for male,44.7% for female). Overall hepatitis B virus (HBV) infection rate was 49.1% and the infection rate for male (50.3%) was slightly higher than that for female (46.5%). HBsAg positive rate and infection rate were increased as the grade increased but it was attributed to the age distribution of the students. HBaAg positive rate for 20 years old students was 1.7%; 21 years, 6.6%; 22 years, 6.1%; 23 years, 12.2%; and 24 years and older, 6.4%. HBV infection rate showed an increasing trend as age increased; 45.8% for 20 years,41.5% for 21 years, 49.5% for 22 years, 55.5% for 23 years and 59.6% for 24 years and older. The age differences in HBsAg positive rates and HBV infection rates did not reach the statistical significance level of 0.05. However, these findings and similar age differences in HBsAg positive rates and HBV infection rates observed in other study suggest that there is a significant age differences. Study of the same age group in other schools and different social classes is warranted to confirm the age difference. Clarification of the reason for such differences would provide a clue to identify the major route of HBV transmission in this age group. Among 26 HBsAg positive students in the initial test, only one student was active hepatitis patient. Out of 24 students who had follow-up test after 16 months 22 students were positive for HBaAg and two students became HBsAg negative and anti-HBs positive. It is obvious that nearly one-half of the medical school students were infected with HBV before 20 years of age and the HBV infection occurs in medical school. Thus, it is recommended to test all the students for HBV infection soon after the admission to the medical school and immunize all the susceptible students with hepatitis B vaccine and give booster as they start to practice at a hospital.

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Analysis of cytosine adenine repeat polymorphism of the IGF-I promoter gene in children with idiopathic short stature (특발성 저신장증 환자에서 IGF-I 프로모터 cytosine-adenine repeat 유전자 다형성의 분석)

  • Moon, Jae Hoon;Chung, Woo Yeong
    • Clinical and Experimental Pediatrics
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    • v.52 no.3
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    • pp.356-363
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    • 2009
  • Purpose : A polymorphism in the IGF-I gene promoter region is known to be associated with serum IGF-I levels, birth weight, and body length, suggesting that IGF-I gene polymorphism might influence postnatal growth. The present study aimed to investigate the role of this polymorphic cytosine-adenine (CA) repeat of the IGF-I gene in children with idiopathic short stature. Methods : The study involved 131 children (72 boys and 59 girls) diagnosed with idiopathic short stature, aged 715 years. Genomic DNA was extracted from anticoagulated peripheral whole blood. The primers were designed to cover the promoter region containing the polymorphic CA repeat. Data were analyzed using GeneMapper software. The correlations between age and serum IGF-I levels were analyzed using Spearmans correlation coefficient. Results : The CA repeat sequences ranged from 15 to 22, with 19 CA repeats the most common with an allele frequency of 40.6%. Homozygous for 19 CA repeat was 13.0%, heterozygous for 19 CA repeat was 56.5%, and 19 CA non-carrier was 30.5%. The three different genotype groups showed no significant differences in height, body weight and body mass index, and serum IGF-I levels. The serum IGF-I level and age according to the IGF-I genotypes were significantly correlated in the entire group, 19 CA repeat carrier group, and the non-carrier group. The three groups also showed no significant differences in the first year responsiveness to GH treatment. Conclusion : There were no significant different correlations between 19 CA repeat polymorphism and serum IGF-I levels according to genotype. Our results suggest that the IGF-I 19 CA repeat gene polymorphism is not functional in children with idiopathic short stature.

Perinatal Factors Affecting Thyroid Stimulating Hormone(TSH) and Thyroid Hormone Levels in Cord Blood (제대혈 갑상선 자극호르몬과 갑상선 호르몬 농도에 영향을 주는 주산기 인자들)

  • Kim, Eun Young;Park, Sang Kee;Song, Chang Hun;LIm, Sung-Chul
    • Clinical and Experimental Pediatrics
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    • v.48 no.2
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    • pp.143-147
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    • 2005
  • Purpose : The aim of the this study was to evaluate the effect of various perinatal conditions on TSH and thyroid hormone levels in cord blood. Methods : Cord blood samples were collected from 130 neonates immediately after birth. TSH, $T_3$, and free $T_4$ levels were measured by the radioimmunoassay(RIA) method. The effects of gestational age, sex, birth weight, delivery method, perinatal asphyxia, maternal diabetes mellitus(DM), and preeclampsia on TSH and thyroid hormone levels were assessed by ANOVA test, Student t-test, and multiple regression analysis. Results : Birth weight and sex did not affect TSH and thyroid hormone levels. TSH level increased according to gestational age(P<0.05). TSH level was $4.42{\pm}0.66{\mu}IU/mL$ in infants born vaginally, which was higher than that of cesarian section delivery($3.31{\pm}0.33{\mu}IU/mL$)(P<0.05). TSH level was $5.18{\pm}0.93{\mu}IU/mL$ in asphyxiated newborns and $2.97{\pm}0.84{\mu}IU/mL$ in non-asphyxiated newborns(P<0.05). TSH level in infants with maternal DM($8.911{\pm}1.25{\mu}IU/mL$) was higher than that of infants without maternal DM($4.32{\pm}0.42{\mu}IU/mL$)(P<0.05). TSH level was $5.28{\pm}0.42{\mu}IU/mL$ in infants with maternal preeclampsia and $3.65{\pm}0.46{\mu}IU/mL$ in infants without maternal preeclampsia(P<0.05). Thyroid hormones were lower in infants with perinatal asphyxia(P<0.05). In asphyxiated infants, $T_3$ level was $75.33{\pm}55.65ng/mL$ and free $T_4$ was $0.54{\pm}0.21ng/mL$. $T_3$ and free $T_4$ level was $109.85{\pm}41.77ng/mL$ and $0.76{\pm}0.22ng/mL$ each in infants without perinatal asphyxia. Among the perinatal factors, gestational age, 1 min Apgar score and maternal DM influenced TSH level independently. Conclusion : In our study, cord blood TSH and thyroid hormone levels were affected by perinatal stress events.

Study on the diagnostic utility of serum levels of insulin-like growth Factor-I and insulin-like growth factor binding protein-3 in growth hormone deficiency (성장호르몬 결핍증 진단에 있어서 혈중 insulin-like growth factor-I 및 insulin-like growth factor binding protein-3 농도의 진단적 유용성에 대한 연구)

  • Ji, Geun Ha;Lee, Jeong Nyeo;Chung, Woo Yeong
    • Clinical and Experimental Pediatrics
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    • v.51 no.12
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    • pp.1329-1335
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    • 2008
  • Purpose : This study aimed to determine the best cutoff line for insulin-like growth factor (IGF)-I and insulin-like growth factor binding protein (IGFBP)-3 to discriminate between growth hormone deficiency (GHD) patients and the control group. Methods : Two hundred thirty subjects with normal controls (129 boys and 101 girls, aged 7-15 years), 14 patients with complete GHD (12 boys and 2 girls), and 17 patients with partial GHD (9 boys and 8 girls) were studied. IGF-I serum concentrations were measured by radioimmunoassay (RI), and IGFBP-3 concentrations were measured by immunoradiometric assay (IRMA). Results : The receiver operating characteristic (ROC) plot analysis showed that the best IGF-I and IGFBP-3 cutoff line was at -1 standard deviation (SD). By comparing IGF-I serum levels of GHD children within 1 SD of normal control, we determined the sensitivity (S) (87.5-100%) and specificity (Sp) (80-84.6%) according to the age group. For IGFBP-3, we determined the following values: S (58.7-85.7%) and Sp (79.2-85.5%). Eleven of 14 patients with complete GHD (78.5%) and 16 of 17 patients with partial GHD (94.1%) had IGF-I concentrations equal to or below -1 SD of the control group mean. Ten of 12 complete GHD children (83.3%) and 13 of 17 partial GHD children (76.5%) had IGFBP-3 concentrations equal or below -1 SD of the control group mean. Conclusion : We conclude that the measurement of IGF-I and IGFBP-3 concentrations might provide essential supplementary data in the diagnostic evaluation of patients with GHD. Our results support the need to use cutoff lines based on below -1 SD of the control.

The Assessment of Ultrasensitive HBsAg kit's Sensitivity level and Performance in Detection of Mutant Forms (Ultra-sensitive HBsAg IRMA 키트의 민감도 및 변이형 검출능 평가)

  • Ha, Dong-Hyuk;Min, Kyung-Sun;Noh, Gyeong-Woon;Kim, Hyun-Ju
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.1
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    • pp.121-125
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    • 2011
  • Purpose: The DNA-type virus HBV, discovered by D. Dane and others in 1976, is approximately 42nm big and known as the main cause of liver-related diseases around the world. HBsAg has 4 kinds of subtypes including adw, adr, ayw and ayr and besides common antigen factor a, there are d, y, r, w. From the methods of serologically testing HBV, IRMA, EIA and CLIa were developed for testing HBsAg and are being used in examining the surface antigen of HBV. In this study, among the methods for testing HBV, the recently developed RIAKEY Ultrasensitive HBsAg IRMA kit's sensitivity level and performance in detection of mutant forms were measured and compared with CLIA. Materials and methods: Two certified reference materials, which are WHO 1st International Standard 1985(80/549) and WHO 2nd International Standard 2003(00/588. subtype adw2, genotypeA), were used in the examination and the sensitivity level was measured by diluting these materials from 0.08 IU/ml to 0.005 IU/ml. The materials for examining the detection of mutant forms included 9 kinds of subtype 'ad' and one kind of subtype 'ay' purchased from DSI company. Also, with the use of positive and negative samples, they was compared with CLIA. Result: Ultrasensitive HBsAg kit based on IRMA method showed the detection of up to 0.01 IU/ml not only for WHO 1st International Standard 1985(80/549) but also for WHO 2nd International Standard 2003(00/588. subtype adw2, genotypeA) and the sensitivity level was measured as 0.01 IU/ml by WHO standard. In testing the performance for detection of mutant forms, the 9 kinds of subtype 'ad' and one kind of subtype 'ay' mutant materials were detected, demonstrating the capacity of detecting various types of mutant forms. Conclusions: With the clinical importance of sensitivity level and performance in detection of mutant forms increasing in the field of HBsAg diagnosis, the examination of IRMA's effectiveness using RIA method in the aspects of the sensitivity level and performance in detection of mutant forms was carried out and its result is as follows. The sensitivity level was measured as 0.01 IU/ml by WHO standard and it was possible to measure various types of mutant forms with high sensitivity. Thus it is suggested that more speedy and accurate reports could be produced from a nuclear medicine laboratory for clinical practitioners requiring results of various situations.

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