• Title/Summary/Keyword: 분자 진단

Search Result 336, Processing Time 0.024 seconds

Molecular Characteristics of Phytophthora katsurae Using PCR-SSCP Analysis (PCR-SSCP 분석에 의한 Phytophthora katsurae의 분자생물학적 특성)

  • Lee, Sun-Keun;Jang, Ha-Na;Lee, Dong-Hyeon;Lee, Sang-Hyun;Lee, Sang-Yong;Lee, Jong-Kyu
    • Research in Plant Disease
    • /
    • v.17 no.2
    • /
    • pp.169-176
    • /
    • 2011
  • Phytophthora katsurae is the fungus responsible for chestnut ink disease. The objectives of this study were to determine if a single-strand conformation polymorphism (SSCP) analysis of rDNA-ITS region, elongation factor 1 alpha gene and ${\beta}$-tubulin gene could be used for rapid identification and genetic diversity of P. katsurae, and to assess the potential use of the SSCP technique as a diagnostic tool for P. katsurae. Each regions amplified by PCR using primers designed to overlap the genus Phytophthora were characterized for the Phytophthora species. PCR products were denatured and electrophoresed for SSCP analysis. P. katsurae isolates showed an unique pattern in SSCP analysis and were easily distinguished from other Phytophthora species used as the control. This indicates that SSCP analysis is an useful technique for distinguishing Phytophthora species from genetically close relatives, and show that the SSCP analysis of each region is an efficient detection tool for P. katsurae. But PCR-SSCP analysis of single-gene may have difficulty in distinguishing P. katsurae from other Phytophthora species. Therefore, PCR-SSCP analysis of multi-genes can be useful for rapid and effective identification of P. katsurae.

Characteristics of Cucumber mosaic virus-VCH Causing Vein Chlorosis on Red Pepper in Korea (고추에 엽맥퇴록병을 일으키는 오이 모자이크 바이러스(CMV-VCH)의 특징)

  • Cho, Jeom-Deog;Lee, Sin-Ho;Kim, Jeong-Soo;Choi, Gug-Seoun;Kim, Hyun-Ran;Chung, Bong-Nam;Ryu, Ki-Hyun
    • Research in Plant Disease
    • /
    • v.12 no.3
    • /
    • pp.226-230
    • /
    • 2006
  • Cucumber mosaic virus(CMV) was occurred on red pepper showing vein chlorosis or vein necrosis with the incidence rate of 52% from 62 specimens collected in natural fields. Among 32 samples infected with CMV, the specimens of 22 red pepper leaves showing vein chlorosis were infected singly with CMV-VCH. CMV-VCH induced vein chlorosis on the inoculated leaves, and vein banding and vein necrosis on the upper leaves of Nicotiana glutinosa, and then killed after showing bud necrosis. The typical symptoms of vein banding, malformation and blister were produced on the upper leaves of Nicotiana benthamiana and N. tabacum 'Ky-57' without symptoms on the inoculated leaves. The commercial cultivars of 'Bugang', 'Manitta' and 'Gwariput' were shown the typical symptom of vein chlorosis by the mechanical inoculation of CMV-VCH. CMV-VCH was detected specifically by RT-PCR. Virus particles of CMV-VCH were isometric shape having 30 nm diameter. Ultraviolet absorption of purified CMV-VCH was maximum at 260 nm and minimum at 242 nm. The ratio of A260/A280 was 1.71. CMV-VCH had the single nucleo-protein having the molecular weight of 24.5 kDa.

The Exon 2 Deletion of the COMMD1 Causing Copper Toxicosis in Bedlington Terriers in Korea (한국 베들링턴 테리어에서 구리중독증을 유발하는 COMMD1 유전자의 exon 2 결손변이)

  • Kim, Yun-Gi;Kim, So-Yeon;Yun, Young-Min
    • Journal of Veterinary Clinics
    • /
    • v.32 no.1
    • /
    • pp.1-4
    • /
    • 2015
  • This study was performed to survey prevalence of Copper metabolism domain containing 1 (COMMD1) mutation using molecular diagnostic method in a population of Bedlington terriers in Korea. COMMD1 gene (formerly MURR1) functions as a regulator of sodium transport and copper metabolism. The deletion of exon 2 of the COMMD1 gene causes copper toxicosis in Bedlington terriers. Bedlington terriers with this autosomal recessive disorder were shown to have the elevated liver copper levels due to genetic derangement in the biliary copper excretion pathway. DNA samples were extracted from whole blood collected from 257 Bedlington terriers (109 males, 148 females) of pet dog clubs in Korea. A multiplex PCR was carried out to detect of exon 2 deletion of COMMD1 gene. In this study, it was possible to know the existence and prevalence of exon 2 deletion of COMMD1 in Bedlington terriers in Korea. Of the 257 samples, 131 (51%) were wild type homozygous for the normal COMMD1 gene, 108 (42%) were heterozygous, having both normal and mutated copy of the COMMD1 gene. The eighteen (7%) were mutant type homozygous. The results of genetic analysis could help establish proper management strategy and selective breeding program to prevent COMMD1 mutation in Bedlington terriers in Korea.

Comparative Diagnostic Studies on Serologic and Molecular Biological Tests Against Haemorrhagic Fever with Renal Syndrome (신증후출혈열 환자의 혈청학적 및 분자생물학적 진단 검사법 비교)

  • 우영대;문희주;배형준
    • Biomedical Science Letters
    • /
    • v.6 no.2
    • /
    • pp.141-149
    • /
    • 2000
  • The etiologic agents of haemorrhagic fever with renal syndrome (HFRS) in Korea are Hantaan and Seoul virus in the genus Hantavirus, family Bunyaviridae. Antibody titers of sera from HFRS patients against Hantaan virus were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA) and plaque reduction neutralization test (PRNI). PRNT and nested reverse transcriptase polymerase chain reaction (nested RT-PCR) was used for serotypic differentiation of Hantaviruses against Hantaan and Seoul virus. Eight doubtful HFRS patients showed higher fluorescent, IgG ELISA, agglutination and neutralizing antibody titer by IFAT, ELISA IgG, HDPA and PRNT, respectively Five out of them showed high IgM antibody titer by IgM capture ELISA against Hantaan virus, remarkably. Fifteen HFRS patients showed higher fluorescent antibody titer by IFAT. In PRNT, 12 out of them showed high neutralizing antibody titer against HTNV, 2 against SEOV and 1 against both viruses. In nested RT-PCR using serotype specific-primer, 3 out of them showed positive against HTNV and 1 against SEOV.

  • PDF

Production of the Recombinant Single Chain Anti-B Cell Lymphoma Antibody and Evaluation of Immunoreactivity (pET vector를 통한 유전자 재조합 단일사슬 항 B형 림프종 항체의 생산과 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kang-Sun;Chung, Wee-Sup;Kim, Soo-Gwan;Cheon, Gi-Jeong;Choi, Chang-Woon;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
    • /
    • v.40 no.4
    • /
    • pp.211-217
    • /
    • 2006
  • Purpose: Recombinant ScFv lym-1 was produced, using pET vector system for large scale production. Methods: ScFv lym-1 gene inserted pET-22b (+) vector, was expressed in E.coli BL-21 strain. ScFv lym-1 antibody extracted from periplasm, was purified with His-Taq column. To evaluated immunoreactivity with Raji cell, ScFv lym-1 was labeled with I-125 and I-125 ScFv lym-1 was purified with desalting column. Raji cell was injected into the C57BR/cdJ SCID mice. Gamma camera imaging were taken time point at 1, 8, 24, and 48 hr with 8 mm pinhole collimator. Results: An active scFv lym-1 could be produced in E. coli with soluble iron using PET vector system. Immuuoreaetivity and affinity constant of IgG lym-1 were 54% and $1.83{\times}10^9M^{-1}$, respectively, and those of scFv lym-1 were 53.7% and $1.46{\times}10^9M^{-1}$, respectively. Biodistribution of I-125 scFv lym-1 antibody showed faster clearance in blood, spleen, kidney and than I-125 IgG lym-1 antibody. Gamma camera image of I-125 scFv lym-1 antibody showed faster clearance and tumor targeting liver than I-125 IgG lym-1 antibody. Conclusions: In vitro properties of scFv lym-1 were similar to those of IgG lym-1. ScFv lym-1 showed faster blood clearance than IgG lym-1 There results suggest that scFv lym-1 antibody can be useful for tumor imaging agent.

Roles of Prostatic Acid Phosphatase in Prostate Cancer (Prostatic acid phosphatase의 전립선 암에서의 역할)

  • Kong, Hoon-Young;Lee, Hak-Jong;Byun, Jong-Hoe
    • Journal of Life Science
    • /
    • v.21 no.6
    • /
    • pp.893-900
    • /
    • 2011
  • Prostatic acid phosphatase (PAP) is one of the widely used biomarkers in the diagnosis of prostate cancer. It was initially identified in 1935 and is the most abundant phosphatase in the human prostate. PAP is a prostate-specific enzyme that is synthesized in prostate epithelial cells. It belongs to the acid phosphatase group that shows enzymatic activity in acidic conditions. PAP is abundant in prostatic fluid and is thought to have a role in fertilization and oligospermia. It also has a potential role in reducing chronic pain. But one of the most apparent functions of PAP is the dephosphorylation of macromolecules such as HER-2 and PI3P that are involved in the ERK1/2 and MAPK pathways, which in turn leads to inhibition of cell growth and tumorigenesis. Currently, clinical trials using PAP DNA vaccine are underway and FDA-approved immunotherapy using PAP is commercially available. Despite these clinically important aspects, molecular mechanisms underlying PAP regulation are not fully understood. The promoter region of PAP was reported to be regulated by NF-${\kappa}B$, TNF-${\alpha}$, IL-1, androgen and androgen receptors. Here, the features of PAP gene and protein structures together with the function, regulation and roles of PAP in prostate cancer are discussed.

Analysis of the Correlation between Expressions of HSP90α, HSP90β, and GRP94, and the Clinicopathologic Characteristics in Tissues of Non-Small Cell Lung Cancer Patients (비소세포 폐암 환자 조직에서 Hsp90α, Hsp90β, GRP94의 발현과 임상병리학적 특성과의 상관관계 분석)

  • Kim, Mi Kyeong
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.49 no.4
    • /
    • pp.460-469
    • /
    • 2017
  • Heat shock proteins (HSPs) are induced as a self-defense mechanism of cells when exposed to various external stresses, such as high fever, infection, free radicals, and heavy metals. They affect the prognosis in the process of tumor formation. HSP is classified into four families: HSP27, HSP60, HSP90, and HSP100, depending on molecular weight. Heat shock protein 90 (HSP90), a molecular chaperone, plays an important role in the cellular protection against various stressful stimuli and in the regulation of cell cycle progression and apoptosis. In the present study, we assessed the differential expression of HSP90 family proteins in non-small cell lung cancer (NSCLC), and the correlation of their expression levels with clinicopathologic factors and patient survival rates. The result of this study can be summarized as follows; $HSP90{\alpha}$ showed higher expression in patients with no lymphovascular invasion (p=0.014). $HSP90{\beta}$ showed a higher expression of squamous cell carcinoma (p=0.003), and an over expression of glucose-related protein (GRP94) was significantly associated with poor differentiation (p=0.048). However, none of the HSP90 proteins showed a significant association with the survival status in patients with NSCLC. This study also indicates that $HSP90{\alpha}$ might contribute more to the carcinogenesis of NSCLC than $HSP90{\beta}$, and GRP94 and isoform selectivity should be considered when HSP90 inhibitors are studied or utilized in the treatment of NSCLC.

Phylogenetic Analysis of Human Bocavirus in Hospitalized Children with Acute Respiratory Tract Infection in Korea (급성 호흡기 감염으로 입원한 소아에서 분리된 보카바이러스의 계통분석)

  • Ahn, Jong Gyun;Choi, Seong Yeol;Kim, Dong Soo;Kim, Ki Hwan
    • Pediatric Infection and Vaccine
    • /
    • v.19 no.2
    • /
    • pp.71-78
    • /
    • 2012
  • Purpose: Human bocavirus (hBoV), a recently discovered virus, has been detected in children with respiratory tract infections worldwide. The aim of this study was to analyze the frequency and molecular phylogeny of hBoV in the respiratory samples of children with acute respiratory tract infections in 2010. Methods: Nasopharyngeal samples were collected from 953 children with lower respiratory tract infections at Severance children's hospital in Korea from January 2010 to December 2010. We applied the multiplex PCR technique for the identification of 12 respiratory viruses from the samples. Among the total specimens, hBoV positive samples were subjected to phylogenetic analysis by sequencing a fragment of the VP1/VP2 gene junction. Results: hBoV was detected in 141 (14.8%) among 953 patients. The 61.7% of hBoV-positive samples were found to co-exist with other respiratory viruses. The results of phylogenetic analysis showed that all 141 hBoV-positive isolates were identified as hBoV 1, revealing a high similarity among the isolates (>98%). Conclusion: hBoV 1 with minimal sequence variations circulated in children with acute respiratory infections during 2010. More research is needed to determine the clinical severity and outcomes of the minimal sequence variations.

  • PDF

Immunoblot observation of antigenic protein fractions in Paragonimus tvestermani reacting with humall patients sera (폐흡충 항원단백질에 대한 폐흠충증 한자 혈청의 반응 양상)

  • Kim, Sung-Hwan;Kong, Yoon;Kim, Suk-Il;Kang, Shin-Yong;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
    • /
    • v.26 no.4
    • /
    • pp.239-244
    • /
    • 1988
  • In order to observe the antigenic fractions in saline extract of adult Paragonimus westermani, proteins in the crude extract were separated by sodium dodecyl sulfate-polyacylamide gel electrophoresis(SDS.PAGE) in reducing conditions. The separated protein fractions were transferred to nitrocellulose paper on which 20 sera from human paragonimiasis were reacted and immunoblottrd. Out of 15 stained protein bands in SDS-PAGE, 7 reacted with infected sera while 8 did not. Additionally, 7 unstained protein bands in SDS-PAGE reacted with the sera. Of 14 reacted bands, 30 kilodalton(kDa) band was the most frequently reacted (95%) and was a strong antigen. Protein bands of 23 and 46 kDa were also strong antigens. Bands of over 150 kDa, 120 kDa, 92 kDa, 86 kDa, 74 kDa, 62 kDa, 51 kDa, 32 kDa, 28 kDa, 16.5 kDa and 15.5 kDa were also reactive but their frequencies of the reaction were variable. Key words: Paragonimus westermani, human paragonimiasis, antigenic proteins, SDS-PAGE/ immunoblot

  • PDF

A Case of Carbamoyl Phosphate Synthetase 1 Deficiency with a Relatively Good Prognosis Presented in the Late Neonatal Period (신생아 후기에 증상을 발현하여 비교적 양호한 예후를 보이는 Carbamoyl Phosphate Synthetase 1 Deficiency 1례)

  • Park, Esther;Kim, Min-sun;Song, Ari;Im, Min Ji;Jang, Ja-Hyun;Kim, Ji Hye;Cho, Sung Yoon;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
    • /
    • v.18 no.1
    • /
    • pp.23-29
    • /
    • 2018
  • Carbamoyl phosphate synthetase 1 deficiency (CPS1D) is a rare autosomal recessive urea cycle disorder characterized by hyperammonemia. CPS1D is caused by mutations in the CPS1 gene on chromosome 2q35. Based on the age of onset, there are two phenotypes: the neonatal type and the delayed-onset type. The severity of clinical manifestation depends on the degree of CPS1 residual enzymatic activity, and can result in hyperammonemia and neurological dysfunction. We report a case of CPS1D in a neonate who developed vomiting, decreased consciousness and hyperammonemia at 25th day after birth. She showed excellent response to treatment including hydration, ammonia-lowering drugs and a low-protein diet without hemodialysis. Her growth, development and neurological outcomes were fair at the last follow-up at 17 months of age.

  • PDF