• Title/Summary/Keyword: Kappa analysis

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Cloning of a Glutathione S-Transferase Decreasing During Differentiation of HL60 Cell Line (HL6O 세포주의 분화 시 감소 특성을 보이는 Glutathione S-Transferase의 클로닝)

  • Kim Jae Chul;Park In Kyu;Lee Kyu Bo;Sohn Sang Kyun;Kim Moo Kyu;Kim Jung Chul
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.151-157
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    • 1999
  • Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

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Polymorpshisms of XPC Gene and Risk of Primary Lung Cancer in Koreans (한국인에서 XPC 유전자의 다형성과 원발성 폐암의 위험도)

  • Kim, Kyung-Rock;Lee, Su-Yeon;Choi, Jin-Eun;Kim, Kyung-Mee;Jang, Sang-Soo;Jung, Chi-Young;Kang, Kyung-Hee;Jeon, Kyung-Neoyh;Cha, Sung-Ick;Kim, Chang-Ho;Kam, Sin;Jung, Tae-Hoon;Park, Jae-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.2
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    • pp.113-126
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    • 2002
  • Background : DNA repair plays a crucial role in protection from cancer-causing agents. Therefore, a reduced DNA repair capacity can increase the susceptibility to lung cancer. The XPC gene contains 15 exons and encodes a 940 amino acid protein that plays a central role in DNA damage recognition of the nucleotide excision repair pathway, which is a major DNA repair mechanisim removing the bulky-helix distorting DNA lesions caused by smoking. Recently several polymorphisms in the XPC gene were identified. In addition, it is possible that these polymorphisms may affect the DNA repair capacity, which modulate cancer susceptibility. The relationship between codon 499 and 939 polymorphisms, and a poly(AT) insertion/deletion polymorphism in the XPC gene, and the lung cancer risk were investigated. Materials and Methods : The genotypes were determined using either PCR or PCR-RFLP analysis in 219 male lung cancer patients and 150 healthy males controls. Results : The frequencies of the genotypes (Val499Ala, PAT and Lys939Gln) among the cases were not significantly different from those of the controls. There was no significant associantion between these polymorphi는 and the lung cancer risk when the analyses were stratified according to age, smoking status and the pack-years of smoking. Moreover, the genotypes had no apparent relationship with any of the histological types of lung cancer. There was a linkage disequilibrium among the Val499Ala, PAT and Lys939Gln polymorphisms. The PAT polymorphism had a strong linkage disequilibrium with the Lys939Gln polymorphism (kappa value=0.87). The XPC haplotypes showed no significant association with the lung cancer risk. Conclusion : These results suggest that XPC Val499Ala, PAT and Lys939Gln polymorphisms are not major contributors to the individual lung cancer susceptibility in Koreans.

Anti-inflammatory Activity of Antimicrobial Peptide Protaetiamycine 2 Derived from the Protaetia brevitarsis seulensis (흰점박이꽃무지 유래 항균 펩타이드 프로테티아마이신 2의 항염증활성)

  • Lee, Joon Ha;Baek, Minhee;Lee, Hwa Jeong;Kim, In-Woo;Kim, Sun Young;Seo, Minchul;Kim, Mi-Ae;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1218-1226
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    • 2019
  • The white-spotted flower chafer Protaetia brevitarsis seulensis is a medicinally beneficial and important edible insect species. We previously performed an in silico analysis of the Protaetia brevitarsis seulensis transcriptome to identify putative antimicrobial peptides and then tested their antimicrobial and hemolytic activities. These peptides had potent antimicrobial activities against bacteria and yeast without inducing hemolysis. In the present study, the cationic antimicrobial peptide, protaetiamycine 2, was selected for further assessment of its anti-inflammatory properties in mouse macrophage Raw264.7 cells. Protaetiamycine 2 treatment of Raw264.7 cells suppressed LPS-induced nitric oxide production and reduced the expression of inducible nitric oxide synthase and cyclooxygenase-2, as determined by real-time PCR and western blotting. The expression of proinflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$) was also attenuated through the MAPKs and $NF-{\kappa}B$ signaling. We also confirmed that protaetiamycine 2 bound to bacterial cell membranes by a specific interaction with LPS. Collectively, these data obtained from LPS-induced Raw264.7 cells indicated that protaetiamycine 2 could have both antimicrobial and anti-inflammatory properties.

Simultaneous Detection of Seven Phosphoproteins in a Single Lysate Sample during Oocyte Maturation Process (난자성숙 과정의 단일 시료에서 일곱 가지 인산화 단백질의 동시 분석 방법)

  • Yoon, Se-Jin;Kim, Yun-Sun;Kim, Kyeoung-Hwa;Yoon, Tae-Ki;Lee, Woo-Sik;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.3
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    • pp.187-197
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    • 2009
  • Objective: Phosphorylation and dephosphorylation of proteins are important in regulating cellular signaling pathways. Bead-based multiplex phosphorylation assay was conducted to detect the phosphorylation of seven proteins to maximize the information obtained from a single lysate of stage-specific mouse oocytes at a time. Methods: Cumulus-oocyte complexes (COCs) were cultured for 2 h, 8 h, and 16 h, respectively to address phosphorylation status of seven target proteins during oocyte maturation process. We analyzed the changes in phosphorylation at germinal vesicle (GV, 0 h), germinal vesicle breakdown (GVBD, 2 h), metaphase I (MI, 8 h), and metaphase II (MII, 16 h in vitro or in vivo) mouse oocytes by using Bio-Plex phosphoprotein assay system. We chose seven target proteins, namely, three mitogen-activated protein kinases (MAPKs), ERK1/2, JNK, and p38 MAPK, and other 4 well known signaling molecules, Akt, GSK-$3{\alpha}/{\beta}$, $I{\kappa}B{\alpha}$, and STAT3 to measure their phosphorylation status. Western blot analysis and kinase inhibitor treatment for ERK1/2, JNK, and Akt during in vitro maturation of oocytes were conducted for the confirmation. Results: Phosphorylation of ERK1/2, JNK, p38 MAPK and STAT3 was increased over 3 folds up to 20 folds, while phosphorylation of the other three signal molecules, Akt, GSK-$3{\alpha}/{\beta}$, and $I{\kapa}B{\alpha}$ was less than 3 folds. All of these results except for Akt were statistically significant (p<0.05). Conclusion: This is the first report on the new and valuable method measuring many phosphoproteins simultaneously in one minute sample such as oocyte lysates. All of the three MAPKs, ERK1/2, JNK, and p38 MAPK are involved in the process of mouse oocyte maturation. In addition, STAT3 might be important regulator of oocyte maturation, while Akt phosphorylation at Serine 473 may not be involved in the regulation of oocyte maturation.

Detection of Pine Wilt Disease tree Using High Resolution Aerial Photographs - A Case Study of Kangwon National University Research Forest - (시계열 고해상도 항공영상을 이용한 소나무재선충병 감염목 탐지 - 강원대학교 학술림 일원을 대상으로 -)

  • PARK, Jeong-Mook;CHOI, In-Gyu;LEE, Jung-Soo
    • Journal of the Korean Association of Geographic Information Studies
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    • v.22 no.2
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    • pp.36-49
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    • 2019
  • The objectives of this study were to extract "Field Survey Based Infection Tree of Pine Wilt Disease(FSB_ITPWD)" and "Object Classification Based Infection Tree of Pine Wilt Disease(OCB_ITPWD)" from the Research Forest at Kangwon National University, and evaluate the spatial distribution characteristics and occurrence intensity of wood infested by pine wood nematode. It was found that the OCB optimum weights (OCB) were 11 for Scale, 0.1 for Shape, 0.9 for Color, 0.9 for Compactness, and 0.1 for Smoothness. The overall classification accuracy was approximately 94%, and the Kappa coefficient was 0.85, which was very high. OCB_ITPWD area is approximately 2.4ha, which is approximately 0.05% of the total area. When the stand structure, distribution characteristics, and topographic and geographic factors of OCB_ITPWD and those of FSB_ITPWD were compared, age class IV was the most abundant age class in FSB_ITPWD (approximately 55%) and OCB_ITPWD (approximately 44%) - the latter was 11% lower than the former. The diameter at breast heigh (DBH at 1.2m from the ground) results showed that (below 14cm) and (below 28cm) DBH trees were the majority (approximately 93%) in OCB_ITPWD, while medium and (more then 30cm) DBH trees were the majority (approximately 87%) in FSB_ITPWD, indicating different DBH distribution. On the other hand, the elevation distribution rate of OCB_ITPWD was mostly between 401 and 500m (approximately 30%), while that of FSB_ITPWD was mostly between 301 and 400m (approximately 45%). Additionally, the accessibility from the forest road was the highest at "100m or less" for both OCB_ITPWD (24%) and FSB_ITPWD (31%), indicating that more trees were infected when a stand was closer to a forest road with higher accessibility. OCB_ITPWD hotspots were 31 and 32 compartments, and it was highly distributed in areas with a higher age class and a higher DBH class.

Spectral Band Selection for Detecting Fire Blight Disease in Pear Trees by Narrowband Hyperspectral Imagery (초분광 이미지를 이용한 배나무 화상병에 대한 최적 분광 밴드 선정)

  • Kang, Ye-Seong;Park, Jun-Woo;Jang, Si-Hyeong;Song, Hye-Young;Kang, Kyung-Suk;Ryu, Chan-Seok;Kim, Seong-Heon;Jun, Sae-Rom;Kang, Tae-Hwan;Kim, Gul-Hwan
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.23 no.1
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    • pp.15-33
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    • 2021
  • In this study, the possibility of discriminating Fire blight (FB) infection tested using the hyperspectral imagery. The reflectance of healthy and infected leaves and branches was acquired with 5 nm of full width at high maximum (FWHM) and then it was standardized to 10 nm, 25 nm, 50 nm, and 80 nm of FWHM. The standardized samples were divided into training and test sets at ratios of 7:3, 5:5 and 3:7 to find the optimal bands of FWHM by the decision tree analysis. Classification accuracy was evaluated using overall accuracy (OA) and kappa coefficient (KC). The hyperspectral reflectance of infected leaves and branches was significantly lower than those of healthy green, red-edge (RE) and near infrared (NIR) regions. The bands selected for the first node were generally 750 and 800 nm; these were used to identify the infection of leaves and branches, respectively. The accuracy of the classifier was higher in the 7:3 ratio. Four bands with 50 nm of FWHM (450, 650, 750, and 950 nm) might be reasonable because the difference in the recalculated accuracy between 8 bands with 10 nm of FWHM (440, 580, 640, 660, 680, 710, 730, and 740 nm) and 4 bands was only 1.8% for OA and 4.1% for KC, respectively. Finally, adding two bands (550 nm and 800 nm with 25 nm of FWHM) in four bands with 50 nm of FWHM have been proposed to improve the usability of multispectral image sensors with performing various roles in agriculture as well as detecting FB with other combinations of spectral bands.

Analysis of Crustal Velocity Structure Beneath Gangwon Province, South Korea, Using Joint Inversion of Receiver Functions and Surface Wave Dispersion (수신함수와 표면파 분산의 연합 역산을 사용한 강원도 지역 하부의 지각속도구조 분석)

  • Jeong-Yeon Hwang;Sung-Joon Chang
    • Economic and Environmental Geology
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    • v.56 no.3
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    • pp.277-291
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    • 2023
  • To analyze the crustal velocity structures beneath 21 broadband seismic stations in Gangwon Province, South Korea, we first applied the H-κ stacking method to 139 teleseismic event data (Mw ≥ 5.8 and the epicentral distance of 30° - 90°) occurring between March 18, 2019 and December 31, 2022 to estimate the Moho depths and Vp/Vs ratios beneath each station. The Moho depths and Vp/Vs ratios from the H-κ stacking method range from 24.9 to 33.2 km depth and 1.695 - 1.760, respectively, and the estimated Vp/Vs ratios were applied to the joint inversion of receiver functions and surface wave dispersion to obtain 1-D crustal velocity models beneath each station. The resulting Moho depths range from 25.9 to 33.7 km depth, similar to the results from the H-κ stacking method. Moho depth results from the both methods are generally consistent with Airy's isostasy. The 1-D crustal velocity models confirm that the existence of 2 km thick low-velocity layers with P-wave velocities of 5 km/s or less at some stations in the Taebaeksan basin, and at the stations CHNB and GAPB in northern Gangwon Province, which are located above the Cenozoic sedimentary layer. The station SH2B, although not overlying a sedimentary layer, has a low P-wave velocity near the surface, which is probably due to various factors such as weathering of the bedrock. We also observe a velocity inversion with decreasing velocity with depth at all stations within 4 - 12 km depths, and mid-crustal discontinuities possibly due to density differences in the rocks at around 10 km depth below some stations.

THE CHANGE OF BONE FORMATION ACCORDING TO MAGNETIC INTENSITY OF MAGNET PLACID INTO TITANIUM IMPLANT SPECIMENS (타이타늄 임플랜트 시편 내부에 설치한 자석의 자성강도에 따른 골형성 변화)

  • Hwang Yun-Tae;Lee Sung-Bok;Choi Dae-Gyun;Choi Boo-Byung
    • The Journal of Korean Academy of Prosthodontics
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    • v.43 no.2
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    • pp.232-247
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    • 2005
  • Purpose. The purposes of this investigation were to discover the possibility of clinical application in the areas of dental implants and bone grafts by investigating the bone formation histologically around specimen which was depending on the intensity of magnetic field of neodymium magnet inside of the specimens. Material and method. 1. Measurement of magnetic intensity - placed the magnet inside of the specimen, and measured the intensity of magnetic field around the 1st thread and 3rd thread of specimen 20 times by using a Gaussmeter(Kanetec Co., Japan). 2. Surgical Procedure - Male rabbit was anesthetised by constant amount of Ketamine (0.25ml/kg) and Rompun (0.25ml/kg). After incising the flat part of tibia, and planted the specimens of titanium implant, control group was stitched without magnet, while experimental groups were placed a magnedisc 500(Aichi Steel Co., Japan) or magnedisc 800(Aichi Steel Co., Japan) into it, fixed by pattern resin and stitched. 3. Management after the surgery - In order to prevent it from the infection of bacteria and for antiinflammation, Gentamycin and Ketopro were injected during 1 week from operation day, and dressed with potadine. 4. Preparation of histomorphometric analysis - At 2, 4 and 8 weeks after the surgery, the animals were sacrificed by excessed Ketamine, and then, specimens were obtained including the operated part and some parts of tibia, and fixed it to 10% of PBS buffer solution. After embedding specimens in Technovit 1200 and B.P solution, made a H-E stain. Samples width was 75$\mu$m . In histological findings through the optical microscope and using Kappa image base program(Olympus Co. Japan), the bone contact ratio and bone area ratio of each parts of specimens were measured and analyzed. 5. Statistical analysis - Statistical analysis was accomplished with Mann Whitney U-test. Results and conclusion. 1. In histomorphometric findings, increased new bone formation was shown in both control & experimental groups through the experiment performed for 2, 4 & 8 weeks. After 4 weeks, more osteoblasts and osteoclasts with significant bone remodeling were shown in experimental groups. 2. In histomorphometric analysis, the bone contact ratios were 38.5% for experimental group 1, 29.5% for experimental group 2 and 11.9% for control group. Experimental groups were higher than control group(p<0.05) (Fig. 6, Table IV). The bone area ratios were 60.9% for experimental group 2, 46.4% for experimental group 1 and 36.0% for control group. There was no significantly statistical difference between experimental groups and control group(p<0.05) (Fig. 8, Table VII) 3. In comparision of the bone contact ratios at each measurement sites according to magnetic intensity, experimental group 2(5.6mT) was higher than control group at the 1st thread (p<0.05) and experimental group 1 (1.8mT) was higher than control group at the 3rd thread(p<0.05) (Fig. 7, Table V, VI). 4. In comparision of the bone area ratios at each measurement sites according to magnetic intensity, experimental group 2(5.6mT) was higher than control group and experimental group 1 (4.0mT) at the 1st thread(p<0.1) and experimental group 2(4.4mT) was higher than experimental group 1 (1.8mT) at the 3rd thread(p<0.1) (Fig. 9, Table IX, X). Experiment group 2 was largest, followed by experiment group l and control group at the 3rd thread of implant. There was a significant difference at the 1st thread of control group & experiment group 2, and at 1st thread & 3rd thread of experiment group 1 & 2, and not at control group experiment group 1.(p<0.1)

Effects of a Tetramethoxyhydroxyflavone on the Expression of Inflammatory Mediators in LPS-Treated Human Synovial Fibroblast and Macrophage Cells

  • Yoon, Do-Young;Cho, Min-Chul;Kim, Jung-Hee;Kim, Eun-Jin;Kang, Jeong-Woo;Seo, Eun-Hee;Shim, Jung-Hyun;Kim, Soo-Hyun;Lee, Hee-Gu;Oh, Goo-Taeg;Hong, Jin-Tae;Park, Joo-Won;Kim, Jong-Wan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.686-694
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    • 2008
  • The inhibitory effects of 5,6,3',5'-tetramethoxy 7,4'-hydroxyflavone (labeled as p7F) were elucidated on the productions of proinflammatory cytokines as well as inflammatory mediators in human synovial fibroblasts and macrophage cells. p7F inhibited IL-1${\beta}$ or TNF-${\alpha}$ induced expressions of inflammatory mediators (ICAM-1, COX-2, and iNOS). p7F also inhibited LPS-induced productions of nitric oxide and prostaglandin $E_2$ in RAW 264.7 cells. In order to investigate whether p7F would inhibit IL-1 signaling, p7F was added to the D10S Th2 cell line (which is responsive to only IL-1${\beta}$ and thus proliferates), revealing that p7F inhibited IL-1${\beta}$-induced proliferation of D10S Th2 cells in a dose-response manner. A flow cytometric analysis revealed that p7F reduced the intracellular level of free radical oxygen species in RAW 264.7 cells treated with hydrogen peroxide. p7F inhibited IkB degradation and NF-${\kappa}$B activation in macrophage cells treated with LPS, supporting that p7F could inhibit signaling mediated via toll-like receptor. Taken together, p7F has inhibitory effects on LPS-induced productions of inflammatory mediators on human synovial fibroblasts and macrophage cells and thus has the potential to be an anti-inflammatory agent for inhibiting inflammatory responses.

A Recombinant $Plasmodium$ $vivax$ Apical Membrane Antigen-1 to Detect Human Infection in Iran

  • Haghi, Afsaneh Motevalli;Khoramizade, Mohammad Reza;Nateghpour, Mehdi;Mohebali, Mehdi;Edrissian, Gholam Hossein;Eshraghian, Mohammad Reza;Sepehrizadeh, Zargham
    • Parasites, Hosts and Diseases
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    • v.50 no.1
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    • pp.15-21
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    • 2012
  • In Iran, $Plasmodium$ $vivax$ is responsible for more than 80% of the infected cases of malaria per year. Control interventions for vivax malaria in humans rely mainly on developed diagnostic methods. Recombinant $P.$ $vivax$ apical membrane antigen-1 (rPvAMA-1) has been reported to achieve designing rapid, sensitive, and specific molecular diagnosis. This study aimed to perform isolation and expression of a rPvAMA-1, derived from Iranian patients residing in an endemic area. Then, the diagnostic efficiency of the characterized Iranian PvAMA-1 was assessed using an indirect ELISA method. For this purpose, a partial region of AMA-1 gene was amplified, cloned, and expressed in pET32a plasmid. The recombinant $His-tagged$ protein was purified and used to coat the ELISA plate. Antibody detection was assessed by indirect ELISA using rPvAMA-1. The validity of the ELISA method for detection of anti-$P.$ $vivax$ antibodies in the field was compared to light microscopy on 84 confirmed $P.$ $vivax$ patients and compared to 84 non-$P.$ $vivax$ infected individuals. The ELISA cut-off value was calculated as the mean+2SD of OD values of the people living in malaria endemic areas from a south part of Iran. We found a cut-off point of OD=0.311 that showed the best correlation between the sera confirmed with $P.$ $vivax$ infection and healthy control sera. A sensitivity of 81.0% and specificity of 84.5% were found at this cut off titer. A good degree of statistical agreement was found between ELISA using rPvAMA-1 and light microscopy (0.827) by Kappa analysis.