• 제목/요약/키워드: HaeIII

검색결과 309건 처리시간 0.029초

RFLP를 이용한 Gyrodactylus salaris의 internal transcribed spacer(ITS) PCR 위양성 판별 (Determination of false positives in PCR diagnostics based on the internal transcribed spacer (ITS) of Gyrodactylus salaris using RFLP)

  • 김민성;최희정;정지민;권문경;황성돈
    • 한국어병학회지
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    • 제37권1호
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    • pp.147-153
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    • 2024
  • The World Organization for Animal Health (WOAH) recommends two protocols (ITS and COI) for conventional PCR of G. salaris diagnosis. However, ITS PCR protocol may yield false-positive results, leading to unnecessary countermeasures. It's difficult to distinguish between G. salaris and false-positive by similar amplicon size of PCR, since the amplicon size of ITS PCR in G. salaris and false-positive was 1,300 and 1,187 bp, respectively. The nucleotide sequences of ITS false-positive in rainbow trout is 99.7% identical to previously reported host genome sequences of rainbow trout (Oncorhynchus mykiss) and 95.3 to 89.1% identical to those of other salmonid fish species. To reduce false-positive PCR band, PCR was performed by the different annealing temperature, but PCR bands were still detected. In RFLP analysis by HaeIII, the PCR product of G. salaris was digested into four bands of 512, 399, 234 and 154 bp, while the false-positive was digested into seven bands of 297, 263, 242, 144, 93, 80 and 68 bp. In the RFLP patterns digested by HindIII, G. salaris showed two bands of 659 and 640 bp, while false-positive had one fragment of 1,187 bp without any digestion. Therefore, the RFLP method of ITS PCR with HaeIII and HindIII can be used for differentiation between G. salaris and false-positive. These results might provide important information on the improvement of PCR diagnostic method of G. salaris.

측모 두부방사선 계측법에 의한 혼합 치열기 아동의 연조직에 관한 연구 (A ROENTGENOCEPHALOMETRIC STUDY ON THE SOFT TISSUE OF THE CHILDREN IN MIXED DENTITION)

  • 김선해;서정훈
    • 대한치과교정학회지
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    • 제15권2호
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    • pp.229-237
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    • 1985
  • The Purpose of this study was to investigate the differences in soft tissue characteristics according to the dental or skeletal dysplasia. For this purpose, lateral cephalogram of 153 children (Hellman dental age IIIB: control group 32, Angle CIII. div. 1 malocclusion group 55, Angle Cl III group 66) were traced and measured. For these measurements, following conclusions were made. 1. FH A, FH Sn, FH UL, AA' of the Class III group were thicker than those of the normal and Class II group, but FH B, FH LL, BB' of the Class III group were not significantly different from those of the normal group. 2. FH B, FH LL, BB' of the Class II group were thicker than those of the normal and Class III group, but FH A, FH Sn, FH UL, AA' of the Class II group were not significantly different from those of the noraml group. 3. Ans-Sn, FH P were not significantly different in three groups, while PP' of the Class III group was thicker than those of the other groups. 4. The lower lips of the Class II group were more anterioly everted with respect to the lower incisor inclination than those of the other groups. 5. The severity of skeletal dysplasia was partly camouflaged by the soft tissue.

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고래회충유충증 감별 진단을 위한 18S ribosomal DNA (rDNA) PCR-RFLP 법 적용 (Application of the 18S Ribosomal DNA (rDNA) PCR-RFLP Technique for the Differential Diagnosis of Anisakidosis)

  • 김선미;조민경;유학선;차희재;옥미선
    • 생명과학회지
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    • 제19권9호
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    • pp.1328-1332
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    • 2009
  • 고래회충유충증은 해산어류에 기생하는 고래회충과(family Anisakidae)에 속하는 선충류 유충에 의한 질병으로 유충의 직접적인 위장관내 침입으로 인한 병변과 더불어 유충의 분비 배설물에 의한 알레르기 질환도 유발될 수 있다. 고래회충유충증은 A. simplex를 비롯하여 Contracaecum, Pseudoterranova, Hysterothylacium 등의 유충에 의해 야기될 수 있으나 이들에 대한 형태학적 감별 진단은 유충의 형태적 유사성으로 인하여 매우 어려운 경우가 많다. 이러한 형태학적 진단의 어려움을 극복하고 분자생물학적 감별진단 방법을 확립하기 위하여 A. simplex, Contracaecum type A. type C' 및 Goezia 유충을 숭어, 도다리, 고등어, 아나고, 참돔 등 5종의 어류에서 분리하였다. 각각의 유충으로부터 분리한 18S rDNA를 PCR로 증폭한 후 Taq 1, Hinf I, Hha I, Alu 1, Dde I, Hae III, Sau 96I, Sau 3AI 등 8종의 제한효소를 사용하여 PCR-RFLP를 시행하였다. PCR product의 크기는 약 2.0 Kb였으며 Hinf l, Alu 1, Hha I, Dde 1 및 Hae III로 A. simplex와 Contracaecum type C'을 구분할 수 있었다. 그러나 Contracaecum type A의 경우에는 Taq I, Hinf I, Alu I 및 Dde I의 경우에는 2가지 패턴으로 나타났으며 이들 가운데 일부는 A. simplex, Contracaecum type C', 및 Goezia와 동일한 분석 패턴을 보이기도 하였다. Goezia는 사용한 8개의 제한 효소 모두에서 A. simplex 및 Contracaecum type A 및 type C'과 각기 다른 양상을 보였다. 이러한 결과로 18S rDNA PCR-RFLP 방법은 A. simplex와 Contracaecum type C'의 감별 진단에 유용한 것으로 밝혀졌으며, Contracaecum type A의 분류에는 제한적으로 사용되어야 함은 물론 형태학적 분류 기준에 대한 재검토가 뒤따라야 할 것으로 사료되었다.

중환자 중증도 평가도구의 타당도 평가 - APACHE III, SAPS II, MPM II (Comparing the Performance of Three Severity Scoring Systems for ICU Patients: APACHE III, SAPS II, MPM II)

  • 권영대;황정해;김은경
    • Journal of Preventive Medicine and Public Health
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    • 제38권3호
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    • pp.276-282
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    • 2005
  • Objectives : To evaluate the predictive validity of three scoring systems; the acute physiology and chronic health evaluation(APACHE) III, simplified acute physiology score(SAPS) II, and mortality probability model(MPM) II systems in critically ill patients. Methods : A concurrent and retrospective study conducted by collecting data on consecutive patients admitted to the intensive care unit(ICU) including surgical, medical and coronary care unit between January 1, 2004, and March 31, 2004. Data were collected on 348 patients consecutively admitted to the ICU(aged 16 years or older, no transfer, ICU stay at least 8 hours). Three models were analyzed using logistic regression. Discrimination was assessed using receiver operating characteristic(ROC) curves, sensitivity, specificity, and correct classification rate. Calibration was assessed using the Lemeshow-Hosmer goodness of fit H-statistic. Results : For the APACHE III, SAPS II and MPM II systems, the area under the receiver operating characterist ic(ROC) curves were 0.981, 0.978, and 0.941 respectively. With a predicted risk of 0.5, the sensitivities for the APACHE III, SAPS II, and MPM II systems were 81.1, 79.2 and 71.7%, the specificities 98.3, 98.6, and 98.3%, and the correct classification rates 95.7, 95.7, and 94.3%, respectively. The SAPS II and APACHE III systems showed good calibrations(chi-squared H=2.5838 p=0.9577 for SAPS II, and chi-squared H=4.3761 p=0.8217 for APACHE III). Conclusions : The APACHE III and SAPS II systems have excellent powers of mortality prediction, and calibration, and can be useful tools for the quality assessment of intensive care units(ICUs).

A Simple PCR-RFLP for Idenficiation of Bursaphelenchus spp. Collected from Korea

  • Han, Hye-Rim;Han, Bo-Young;Chung, Yeong-Jin;Shin, Sang-Chul
    • The Plant Pathology Journal
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    • 제24권2호
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    • pp.159-163
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    • 2008
  • Accurate identification of pine wood nematode, Bursaphelenchus xylophilus is a prerequisite to diagnose the pine wilt disease. However, a fungivorous nematode, B. mucronatus is highly similar to B. xylophilus and it is difficult to differentiate these two species by morphological features. A molecular diagnosis method, ITSRFLP was applied for the identification of B. xylophilus and B. mucronatus from Korea. Genomic DNA was extracted from a single individual nematode and ITS DNA was amplified by PCR. The size of PCR product was approximately 900bp and the sequence data were obtained after cloning. Amplified ITS was digested by 5 different restriction enzymes (Rsa I, Hae III, Msp I, Hinf I, and Alu I) and provided a discriminatory profile for B. xylophilus and B. mucronatus. Besides, B. mucro- natus was determined to have 2 different genotypes, East Asian type and European type also clearly separated by Rsa I and Hae III digestion. European type of B. mucronatus is recently collected from Pinus koraiensis and has not been reported before. ITS sequnce data were analyzed by Restriction Mapper program and the result supported ITS-RFLP pattern. These data indicated that PCRRFLP method is an accurate and simple way for identification of Bursaphelenchus species.

A Study on the DNA Polymorphisms at $\beta$fibrinogen Loci and Plasma Fibrinogen Concentration

  • Lee, Mi-Hwa;Ohrr, Hee-Choul;Song, Kyung-Soon
    • 대한의생명과학회지
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    • 제7권3호
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    • pp.151-159
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    • 2001
  • Many prospective studies have confirmed the predictive value of plasma fibrinogen levels for vascular diseases, including ischemic heart disease. Several polymorphisms of the $\beta$fibrinogen gene have been investigated in relation to plasma fibrinogen levels. The blood samples for DNA were collected from 109 healthy Koreans who have no relationship by blood (67 males and 42 females) in due consideration of some other factors such as gender, age, and smoking status. Four polymorphisms of the $\beta$fibrinogen gene that consist of HaeIII, AluI, MaII and BcII restriction fragment length polymorphisms (RFLPs) were investigated to examine the associations between RFLPs and plasma fibrinogen levels. In conclusion, the significant associations between HaeIII, AluI, MnII RFLPs(H$_1$H$_2$, M$_1$M$_2$, $A_1$A$_2$) and the concentration of plasma fibrinogen were shown by the smokers as well as by the old people more than 50, whereas the association between BcII and plasma fibrinogen were shown no connection with the status of age and smoking. The concentration of plasma fibrinogen was significantly shown higher by the old people ($\geqq$50) by the younger people ($\leqq$49) in male and also higher by the smokers than by the nonsmokers.

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Phycocyanin locus내의 DNA Polymorphism에 의한 한국산 Cyanobacteria의 유전적 다양성 (Genetic Diversity of Korean Cyanobacteria determined by DNA polymorphisms within the Phycocyanin Locus)

  • 박진숙;권주리;유순애
    • 미생물학회지
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    • 제36권4호
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    • pp.249-253
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    • 2000
  • Cyanobacteria의 광합성 보조색소인 phycocyanin의 PC operon(cpc gene)을 PCR로 증폭하고, 제한효소로 처리하여 RFLP pattern을 비교하였다. Intergenic spacer sequence를 포함한 cpc gene은 실험에 사용한 cyanobacteria 균주 모두에게 증폭되었으며, 산물의 size는 약 700 bp였다. PCR산물을 5종의 제한효소로 처리한 결과 AluI, MspI, HaeIII는 같은 속내으ㅐ 균주간에 동일한 pattern을 나타내어 속 구분이 가능하였으며 CfoI은 Anabeana와 Synechocystis속의 균주간에 구별되는 양상을 나타내어 속내 균주 구별에 유용하였다. Restriction enzyme profile에 의한 phenogram에서 Anabeana, Chlorogloea, Synechyhocystis는 각각 하나의 cluster를 형성하여 cyanobacteria의 분류에 PC-IGS의 RFLP pattern이 유용함을 알 수 있었다.

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종 특이 PCR과 PCR-RFLP를 이용한 웅담과 기타 담류의 감별 방법 (Identification of Fel ursi and Cattle and Pig Bile Juices by speciesspecific PCR and PCR-RFLP)

  • 권기록;백승일;최석호
    • 대한약침학회지
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    • 제12권1호
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    • pp.13-20
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    • 2009
  • Objective : This study developed species-specific PCR and PCR-RFLP to detect the adulteration of Fel ursi products with cattle and pig bile juices. Methods : All the primers for PCR and PCR-RFLP in this study were designed based on nucleotide sequences of cytochrome b genes in the mitochondria. Results : The species-specific PCR amplified a DNA fragment of 214, 214, 295, and 167 bp from Fel ursi product, bear fur, cattle bile juice, and pig bile juice, respectively. The survey using the speciesspecific PCR indicated that some of commercial Fel ursi products were adulterated with cattle and pig bile juices. PCR-RFLP using the restriction endonucleases, HaeIII and HinfI enabled differentiation among Fel ursi product, cattle bile juice, and pig bile juice. Bear furs from two animals showed variations in PCR-RFLP patterns with HaeIII. Discussion : The detection methods of the species-specific PCR and PCR-RFLP could be useful in eliminating adulterated Fel ursi products from the market.

ELID를 이용한 초미립 숫돌의 원통내면연삭 (Internal Cylindrical Grinding with Super Abrasive Wheel and Electrolytic In- process Dressing)

  • Jun Qian;Gyung Nyun Kim;Hitoshi Ohmori;Hae Do Jeong
    • 한국정밀공학회지
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    • 제17권4호
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    • pp.155-162
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    • 2000
  • 전해 인프로세스 드레싱(ELID)의 응용기술로써 간헐적 드레싱(ELID II) 및 무전극 드레싱(ELID III)이 원통내면 마무리 연삭에 이용되고 있다. 주철본드(CIB-D) 및 메탈레진본드 다이아몬드 숫돌(HRB-D)이 이 방식들에 사용되고 있다. 경면 가공에 있어서 이 두방식은 미립의 숫돌이용으로 일반연삭기에 정밀부속 장치의 보완없이 이용될 수 있다. ELID II 연삭에서 CIB-D숫돌은 파이프 형상의 전극에 의하여 간헐적으로 드레싱되고, 반면에 MRB-D 숫돌은 인프로세스 드레싱 되며 전극은 필요로 하지 않는다. 본 연구에서는 ELID II 및 ELID III 방식에 있어서, 연삭조건 및 연삭입자크기에 대한 연삭특성을 비교검토 하였다. 그 결과, ELID II, III방식 공히 대단히 작은 표면거칠기를 갖는 경면이 얻어짐을 확인하였다.

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