• Title/Summary/Keyword: 중합효소 연쇄반응

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Cigarette Smoking and Polymorphism of the Paraoxonase 1 Gene as Risk factors for Lung Cancer (폐암발생의 위험인자로서 흡연과 Paraoxonase 1 유전자 다형성)

  • Lee, Chul-Ho;Lee, Kye Young;Hong, Yun-Chul;Choe, Kang-Hyeon;Kim, Yong-Dae;Kang, Jong-Won;Kim, Heon;Hong, Jang Soo
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.5
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    • pp.490-497
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    • 2005
  • Background : The paraoxonase enzyme plays a significant role in the detoxification of various organophosphorous compounds in mammals, and paraoxonase (PON) 1 is one of the endogenous free-radical scavenging systems in the human body. In this study, we investigated the interaction between cigarette smoking and the genetic polymorphism of PON1 with lung cancer in Korean males. Methods : Three hundred thirty five patients with lung cancer and an equal number of age-matched controls were enrolled in this study. Every subject was asked to complete a questionnaire concerning their smoking habits and alcohol drinking habits. A 5' exonuclease assay (TaqMan) was used to genotype the PON1 Q192R polymorphism. The effects of smoking habits and drinking habits, the PON1 Q192R polymorphism and their interactions were statistically analyzed. Results : Cigarette smoking and the Q/Q genotype of PON1 were significant risk factors for lung cancer. Individuals carrying the Q/Q genotype of PON1 were at a higher risk for lung cancer as compared with those individuals carrying the Q/R or R/R genotype (odds ratio, 2.84; 95% confidence interval, 1.69 - 4.79). When the groups were further stratified by the smoking status, the Q/Q PON1 was associated with lung cancer among the current or ex-smokers (odds ratio, 2.56; 95% confidence interval, 1.52 - 4.31). Current smokers or ex-smokers who had the Q/Q genotype showed an elevated risk for lung cancer (odds ratio: 15.50, 95% confidence interval: 6.76 - 35.54) as compared with the group of subjects who never smoked, and had the Q/R or R/R genotype. The odds ratios (95% confidence interval) of smokers with the PON1 Q/Q type compared to the nonsmokers with the PON1 Q/R or R/R type were 53.77 (6.55 - 441.14) for squamous cell carcinoma, 6.25 (1.38 - 28.32) for adenocarcinoma, and 59.94 (4.66 - 770.39) for small cell carcinoma, and these results were statistically significant. Conclusion : These results suggest that cigarette smoking and the PON1 Q/Q genotype are risk factors for lung cancer. The combination of cigarette smoking and the PON1 Q/Q genotype significantly increased the lung cancer risk irrespective of the histologic type of cancer.

Seasonal Distribution of T Serotyping and emm Genotyping of Group A Streptococci Obtained from Children with Streptococcal Infections in Masan, Korea, 2003~2004 (2003~2004년 경남 마산 지역에서 분리된 A군 연쇄구균의 T 혈청형과 emm 유전자형의 계절별 분포)

  • Jeon, Ho-Sang;Park, Hwa-Jin;Lee, Hee-Joo;Ma, Sang-Hyuk;Cha, Sung-Ho
    • Pediatric Infection and Vaccine
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    • v.12 no.1
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    • pp.52-60
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    • 2005
  • Purpose : The aim of this study is to know seasonal distribution of group A streptococci obtained from one center using emm genotyping and T serotyping in Masan from 2003 through 2004. Methods : Among children who visited the Changwon Fatima Hospital at Masan, Korea from June 2003 through February 2004, 100 patients who had clinical findings of acute pharyngitis, scarlet fever, and cellulitis were confirmed as GAS by culture, and were enrolled in our study. All obtained GAS were sent to the WHO Collaborative Center for Reference and Research on Streptococci, University of Minnesota, Minneapolis for T serotyping and emm genotyping. We classified these results again according to seasonal and disease's entities. Results : 19 different T serotypes was typed. T4(27.5%), T1(17.6%), T6(13.7%), and T12(13.7%) serotypes were relatively common in summer, while T4(28.3%), T12(15.2%), and T12/B3264(8.7%) were common in winter. T4 and T12 were persistent all year around. Distribution of T serotypes in 89 patients with pharyngotonsillitis were T4(26.7%), T12(14.0%), T1(12.8%), and T6(11.6%) in order of frequency. 15 different emm genotypes was typed. The number of emm 1, emm 6, emm 9, and emm 44 genotypes decreased or disappeared in winter, and the number of emm 3, emm 12, and emm 89 genotypes increased or reappeared in winter. Conclusion : Because T serotyping and emm genotyping are useful tools for evaluating epidemiology and pathogenesis of group A streptococci, we should monitor these strains every year, and should serotype and genotype GAS obtained from the invasive streptococcal infections.

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Prevalence of Periodontopathogens in Saliva and Plaque of Korean Children and Adolescents (한국 소아·청소년의 타액과 치태 내 치주 병원균 출현율)

  • Choi, Hyejin;Kim, Jaehwan;Lee, Daewoo;Yang, Yeonmi;Kim, Jaegon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.43 no.1
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    • pp.8-16
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    • 2016
  • Early colonization of periodontal pathogens has been related as a risk indicator for the subsequent development of periodontal disease. Such colonization can be easily detected with mediums like saliva and plaque. This study aimed to evaluate the prevalence of the bacteria associated with periodontal disease in saliva and plaque in healthy children and adolescents. The experiment was conducted using 90 samples from subjects consisting of thirty elementary school students, thirty high school students and thirty adults. PCR was used to detect the prevalence and distribution of five periodontal pathogens in the collected saliva and plaque. The detected periodontal pathogens are as follows: A. actinomycetemcomitans, P. gingivalis, T. forsythia, F. nucleatum and P. intermedia. Periodontal pathogens were prevailed in a higher number of adolescents than the number of children. A. actinomycetemcomitans and P. intermedia were detected the most in the adolescents group. T. forsythia and F. nucleatum were detected the most in the children group. The overall result showed that saliva is more a useful medium than supragingival plaque. The detection of high risk periodontal pathogens in children and adolescents without clinical signs of periodontal disease can emphasize the importance of the early diagnosis and preventive approach.

Helicobacter pylori vacA Mosaicism and New Primers for vacA Signal Sequence Indigenous to Korea (Helicobacter pylori vacA 대립유전자의 Mosaicism과 Signal Sequence의 한국고유 시발체)

  • Ahn, Yeon-Hwa;Kim, Heung-Ryel;Lee, Ji-Eun;Hwang, Tae-Sook;Choe, Yon-Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.4 no.2
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    • pp.155-160
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    • 2001
  • Purpose: Helicobacter pylori has been known to have diverse vacA allelic types. The purpose of the study was to identify vacA diversity in Korea and design new primers for signal sequence alleles indigenous to Korea. Methods: Fifty antral biopsy specimens, which had been proven to be H. pylori-positive, were examined for vacA status; signal sequence and mid-region. After PCR amplification and DNA sequencing, vacA alleles of Korean H. pylori strains were compared with those from other countries. Results: Among Korean H. pylori strains vacA alleles with all combinations of signal sequence and mid-region were found, with the exception of s1b or s2. vacA genotype s1c/m1 was predominant in Korea. We found that GGGAGCGTTR in s1a and GGGGYTATTG in s1c were the indigenous sequences to Korea and constructed the new Korean specific primers for the vacA signal sequence; VASK-F, VASK-R, S1AK-F, and S1CK-F. Conclusion: This study showed that s1c/m1 is the predominant type of vacA allele in Korea. We designed new primers for the vacA signal sequence.

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DNA barcoding of Raptor carcass collected in the Paju city, Korea (파주시에서 수집한 폐사체 맹금류의 DNA 바코드 연구)

  • Jin, Seon-Deok;Paik, In-Hwan;Lee, Soo-Young;Han, Gap-Soo;Yu, Jae-Pyoung;Paek, Woon-Kee
    • Korean Journal of Environment and Ecology
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    • v.28 no.5
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    • pp.523-530
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    • 2014
  • One juvenile raptor which was not able to be identified due to its head damage was discovered on a roadside in Janggok-ri, Jori-eup, Paju on 28th June, 2011. The species was identified by DNA barcoding. After polymerase chain reaction (PCR) of the mitochondrial cytochrome c oxidase subunit I gene (COI), we obtained 695 bp sequences. We analyzed the obtained COI sequence with similar sequences from the BOLD systems and BLAST of the NCBI Genbank, and discovered that its sequence showed 100 % similarity values with the one of the five gray-faced buzzards which were previously researched. In addition, it was confirmed to be a female through sex determination using DNA. Such results are important information as it confirms the breeding of the gray-faced buzzards for the first time in 43 years since its breeding was last recorded in 1968, in Paju. Wildlife rescue center needs to work with adjacent consigned registration and preservation institutions when carcass of wild animals is collected or DNA samples are obtained for more accurate both species and sex identification through a systematic management system in the future. Furthermore, the obtained DNA sample of the gray-faced buzzard and COI gene, DNA barcode, could be used as reference standards for similar researches in the future.

Validity of the New Caries Activity Test using Real-Time Polymerase Chain Reaction (실시간 중합효소연쇄반응 방법을 이용한 새로운 치아우식 활성 검사법의 유효성)

  • Kwon, Doyoun;Kim, Heejin;Nam, Okhyung;Kim, Misun;Choi, Sungchul;Kim, Kwangchul;Lee, Hyoseol
    • Journal of the korean academy of Pediatric Dentistry
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    • v.45 no.3
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    • pp.354-362
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    • 2018
  • Periogen is a new caries activity test using real-time polymerase chain reaction. The aim of this study was to assess the validity of Periogen by evaluating the correlation with dmft, dmfts indices and comparing with Cariview and caries risk assessment tool (CAT). 83 children under 6 participated in this study. Dmft, dmfts indices and CAT were collected through an examination of oral health status. Plaque samples for Periogen and Cariview were collected and manipulated according to the manufactures' instructions. The correlation coefficient of Periogen, Cariview and CAT with the dmfts index were 0.38, 0.56 and 0.66 in each (p < 0.01). The sensitivity of Periogen, Cariview and CAT were 43%, 76% and 95% and specificity were 80%, 72% and 74% respectively. Area under curve under the receiver operating characteristic curves in each method indicated 0.69, 0.81 and 0.85. CAT and Cariview were more effective in evaluation the risk of dental caries than Periogen so far. To be used Periogen clinically, more improvements for higher validity were needed.

The Prevalence of Plasmid-Mediated Quinolone Resistance Genes among CTX-M-14 Producing Escherichia coli Strains Isolated from a University Hospital in the Chungcheong Province (충청지역에 위치한 일개의 대학병원에서 분리된 CTX-M-14형 ESBL 생성 대장균을 대상으로 PMQR 유전자 빈도조사)

  • Sung, Ji Youn
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.210-216
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    • 2016
  • Recently, there has been a considerable increase in the prevalence of CTX-M type extended-spectrum ${\beta}$-lactamase (ESBL)-producing E. coli isolates worldwide, including Korea. To investigate the ESBL genes in the E. coli strains isolated from a university hospital in the Chungcheong area, a study was conducted using PCR amplification and nucleotide sequence analysis of the amplified products to detect the plasmid mediated quinolone resistance (PMQR) genes in ESBL producing E. coli isolates. The number of CTX-M-14 producing isolates was 25 (16.0%) isolates, and of them, 9 (5.8%) isolates also produced CTX-M-15. All CTX-M type ESBL producing E. coli isolates showed resistance to cefotaxime. Twelve (48%) CTX-M type ESBL producing E. coli isolates contained the PMQR genes, 8 contained qnrS1, and 8 contained aac(6')-Ib-cr. Four isolates harbored both qnrS1 and aac(6')-Ib-cr genes. In our study, we confirmed that the plasmid mediated antimicrobial resistant determinants-the ESBL and PMQR genes-were distributed in the E. coli isolates. To prevent further spreading of the resistant genes among the E. coli isolates, consistent effort is required to investigate and monitor the resistant genes.

A STUDY ON THE IDENTIFICATION OF Porphyromonas endodontalis BY PCR USING SPECIES SPECIFIC PRIMERS FOR THE 16S rDNA (16S rDNA sequence에 대한 종특이성 primer를 이용한 중합효소연쇄반응증폭에 의한 Porphyromonas endodontalis의 동정에 관한 연구)

  • Eom, Seung-Hee;Lim, Sung-Sam;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.13-25
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    • 1999
  • P. endodontalis which was known to be associated with the infected root canals and periapical lesions is very difficult to detect by culture methods or traditional methods. Detection of bacteria using polymerase chain reaction(PCR) for 16S ribosomal DNA(rDNA) is fast, simple, and accurate with relatively small amount of target cells. 16S rDNA consist of conserved regions those are same to all species, and variable regions which represent species specificity. The 16S rDNA sequences of P. endodontalis and P. gingivalis were aligned and two highly variable regions were selected as a pair of species specific oligonucleotide primers for P. endodontalis. And then the pair of primers for PCR amplification was synthesized to identify P. endodontalis. The sequences of the species specific primers for the 16S rDNA of P. endodontalis were as follows ; sense primer[endo1]: 5'-CTATATTCTTCTTTCTCCGCATGGAGGAGG-3' antisense primer[endo2]: 5'-GCATACCTTCGGTCTCCTCTAGCATAT-3' In this study, for the identification of P. endodontalis without culture from the mixed clinical samples, PCR was done with species specific primers for the 16S rDNA sequences of P. endodontalis. The results were as follows : 1. The species specificity of the primers for the 16S rDNA of P. endodntalis was determined by the PCR methods. About 490bp amplicon which was specific only for P. endodntalis was produced with P. endodontalis. No amplicon was produced by PCR with other strains similar to P. endodontalis. 2. The synthesized species specific primers reacted with conventionally identified P. endodontalis which we have in conservative dentistry laboratory. 3. The identification of P. endodontalis using PCR technique with samples collected from infected root canals or periapical lesions was more sensitive than that of culture methods. 4. Seven samples revealed including P. endodontalis by PCR technique. Five of them were related with pains, two of them with sinus tract, three of them with foul odor, and three of them with purulent drainage. P. endodontalis was shown to have great relation with pains.

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IDENTIFICATION OF FUSOBACTERIUM NUCLEATUM AND FUSOBACTERIUM NECROPHORUM USING POLYMERASE CHAIN REACTION(PCR) (중합효소연쇄반응(Polymerase Chain Reaction)을 이용한 Fusobacterium nucleatum 및 Fusobacterium necrophorum의 동점에 관한 연구)

  • Kang, Chang-Woo;Park, Dong-Sung;Yoon, Soo-Han
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.40-48
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    • 1999
  • This study was designed to examine the specificities of the designed primers for F. nucleatum and F. necrophorum and to compare the PCR results using clinical samples with those of the anaerobic culture method. F. nucleatum and F. necrophorum spp. are frequently isolated in infected root canals, and they are related to periapical diseases. F. nucleatum(VPI 10197) and F. necrophorum(ATCC 25286) were used as references for PCR reaction, and thirty five teeth with one canal and periapical lesion were used. The samples were cultured anaerobically and identified using Rapid ID 32A(BioMerieux Vitek, Inc., France) as biochemical battery. In the GenBank database, species-specific PCR primers(nuc1/nuc2 primers for F. nucleatum and nec1/nec2 primers for F. necrophorum) were designed from the 16S ribosomal DNA(rDNA) sequences of F. nucleatum(accession number M58683) and F. necrophorum(accession number AF044948). PCR procedures of F. nucleatum(VPI 10197) and F. necrophorum (ATCC 25286) were simulated on a computer software. Amplify(v.1.2${\beta}$ for Macintosh). 820 bps and 817 bps of nucleotides were expected, respectively. Using extracted DNAs with QiaAmp tissue kit(Qiagen co., Germany), PCR was done. The results were as follows : 1. The nuc1/nuc2 primers produced an amplicon of 820 bps and the nec1/nec2 primers produced an amplicon of 817 bps. 2. The nuc1/nuc2 primers and the nec1/nec2 primers were specific and did not react with species other than the designated ones(i.e. nuc1/nuc2 primers did not produce amplicons for F. necrophorum, and vice versa.). And the PCR products of Porphyromonas endodontalis(ATCC 35406), Porphyromonas gingivalis(ATCC 33277), Prevotella intermedia(ATCC 25611), and Prevotella nigrescens(ATCC 33563), frequently isolated in infected root canals and periapical lesions, were not amplified by the primers specific for Fusobacterium nucleatum and Fusobacterium necrophorum. 3. This method utilizing PCR could detect F. nucleatum and F. necrophorum in clinical samples, while anaerobic culture method could detect neither.

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A Single Nucleotide Deletion resulting in Frameshift in Two Korean Neonates with Thyroxine-Binding Globulin Deficiency (단일 뉴클레오타이드 결손으로 인한 Frameshift 돌연변이로 규명된 티록신결합글로불린 결핍증 1례)

  • Park, Sang-Joon;Suh, Jin-Soon;Jung, Min-Ho;Lee, Hee-Jin;Suh, Byung-Kyu;Lee, Won-Bae;Lee, Byung-Churl
    • Clinical and Experimental Pediatrics
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    • v.48 no.11
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    • pp.1252-1255
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    • 2005
  • Abnormalities in the levels of thyroxine-binding globulin (TBG) are not associated with clinical disease and they do not require treatment. Congenital TBG deficiency is inherited in an X-linked manner. To date, some complete and partial TBG variants and one polymorphism have been identified by analysis of the TBG gene. Two male neonates were referred to us because of their low $T_4$ levels that were noted on the neonatal screening test. They showed normal levels of free $T_4$ and TSH. Their serum TBG was not detectable and those values of their parents were within the normal ranges. The genomic DNA was extracted from their white blood cells and the four coding exons of the TBG gene were amplified by using polymerase chain reaction. Sequencing of the four coding regions and all the intron/exon junctions revealed a single nucleotide deletion of the first base of the codon 352 of the mature protein in both of the neonates. This mutation resulted in a frameshift and a premature stop codon (TGA) 374. Their mothers were shown to be heterozygotes. We detected a single nucleotide deletion resulting in a frameshift in two male Korean neonates who had complete TBG deficiency.