• Title/Summary/Keyword: IS-PCR

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A Current Source using the 12-Pulse Phase-Controlled Rectifier (12-펼스 위상제어 정류기를 사용한 전류원)

  • 송의호;권봉환
    • The Transactions of the Korean Institute of Electrical Engineers
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    • v.39 no.6
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    • pp.545-556
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    • 1990
  • A design method of a current source using 12-pulse phase-controlled rectifier (PCR) is presented. The critical inductance of the 12-pulse PCR is derived and it is shown that the critical inductance can be reduced using a current source. The control circuit of the 12-pulse PCR with an inner fast dynamic loop is proposed to give the frequency synchronism and to reduce the subharmonics due to the unbalance of the transformer of the power line. This circuit is analyzed and its dynamic loop is optimized. The optimal constant PIMF (proportional, integral and measurable variable feedback, and feedforware) controller is also designed using the time-weighted quadratic performance index. It is shown via experimental results that the proposed design method gives high dynamic and static performance of the current source using the 12-pulse PCR.

Comparison of PCR-Line Probe and PCR-SSCP Methods for the Detection of Rifampicin Resistant Mycobacterium Tuberculosis (Rifampicin 내성 결핵균의 검출에 있어서 PCR-line Probe법과 PCR-SSCP법의 비교)

  • Kim, Ho-Joong;Suh, Gee-Young;Chung, Man-Pyo;Kim, Jong-Won;Shim, Tae-Sun;Choi, Dong-Chull;Kwon, O-Jung;Rhee, Chong-H;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.714-722
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    • 1998
  • Background: Rifampicin (RFP) is a key component of the antituberculous short-course chemotherapy and the RFP resistance is a marker of multi-drug resistant (MDR) tuberculosis. RPoB gene encodes the $\beta$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. And the mutations of rpoB gene have been found in about 96% of rifampicin resistant clinical isolates of M. tuberculosis. So in order to find a rapid and clinically useful diagnostic method in identifying the RFP resistance, we compared the PCR -line probe method with PCR-SSCP for the detection of the rpoB gene mutation in cultured M. tuberculosis. Methods: 45 clinical isolates were collected from patients who visited Sung Kyun Kwan University Hospital. The RFP susceptibility test was referred to the referral laboratory of the Korean Tuberculosis Institute. 33 were rifampicin resistant and 12 were rifampicin susceptible. The susceptibility results were compared with the results of the PCR-BSCP and PCR-line probe method. Results: We could find rpoB mutations in 27/33(81.8%) RFP-resistant strains by PCR-line probe method, and in 23/33 (69.7%) by PCR-SSCP and there was no significant difference between two methods. There was no mutation in rifampicinn susceptible strains by both methods. Conclusion: PCR-line probe method would be a rapid, sensitive and specific method for the detection of rifampicin resistant Mycobacterium tuberculosis.

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Quantitative Polymerase Chain Reaction for Microbial Growth Kinetics of Mixed Culture System

  • Cotto, Ada;Looper, Jessica K.;Mota, Linda C.;Son, Ahjeong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1928-1935
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    • 2015
  • Microbial growth kinetics is often used to optimize environmental processes owing to its relation to the breakdown of substrate (contaminants). However, the quantification of bacterial populations in the environment is difficult owing to the challenges of monitoring a specific bacterial population within a diverse microbial community. Conventional methods are unable to detect and quantify the growth of individual strains separately in the mixed culture reactor. This work describes a novel quantitative PCR (qPCR)-based genomic approach to quantify each species in mixed culture and interpret its growth kinetics in the mixed system. Batch experiments were performed for both single and dual cultures of Pseudomonas putida and Escherichia coli K12 to obtain Monod kinetic parameters (μmax and Ks). The growth curves and kinetics obtained by conventional methods (i.e., dry weight measurement and absorbance reading) were compared with that obtained by qPCR assay. We anticipate that the adoption of this qPCR-based genomic assay can contribute significantly to traditional microbial kinetics, modeling practice, and the operation of bioreactors, where handling of complex mixed cultures is required.

Detection of Rice Stripe Virus using RT-PCR (RT-PCR에 의한 벼 줄무늬잎마름병 정밀진단)

  • Lee, Bong-Choon;Hong, Yeon-Kyu;Kwak, Do-Yeon;Oh, Byeong-Geun;Park, Sung-Tae;Kim, Soon-Chul
    • Research in Plant Disease
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    • v.10 no.1
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    • pp.30-33
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    • 2004
  • Until now, occurrence of rice stripe virus (RSV) is limitted in southern part of Korea. However, recently the occurrence of RSV is increased and spreaded in central part of Korea including Chungcheong and Kyonggi province. It is very difficult to distinguish RSV symptoms on virus symptom physiological damage of rice. We detected RSV viral RNA from infected rice and its insect vector Laodelphax striatellus using specific primer of RSV-polymerase and coat protein gene with reverse transcription (RT)-PCR. The result of RT-PCR, we observed specific band including RSV-polymerase (1,,023 bp) and CP (969 bp) in both host of rice and insect vector.

Detection and Identification of $\beta$-lactamase, Enterotoxin and Other Exotoxins Genes of Staphylococcus aureus by PCR

  • Yoon, Y.H.;Kim, K.I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.425-429
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    • 2003
  • Staphylococcus aureus is a major pathogen for cattle, causing various forms of subclinical and clinical mastitis and could be a causative agent of food poisoning, it produces various superantigenic exotoxins which have a great public health significance. A total of 72 S. aureus clinical isolates from dairy farms located in Kyunggi Province Korea were examined for the species identification by biochemical method, and for the detection of $\beta$-lactamase, enterotoxin and other exotoxins genes by PCR. The results of species identification by biochemical method agreed with those of PCR done with species specific primer STA-AU. $\beta$-lactamase is an enzyme closely associated with the resistance to antibiotic penicillin, which is an important means of treatment of mastitis, all the isolates were positive for the presence of genes encoding $\beta$-lactamase, which were reproduced in penicillin susceptibility disc assay. Six types of toxin genes, Staphylococcal enterotoxin (SE)A, SEB, SEC, SEE, toxic shock syndrome toxin (TSST-1) and exfoliative toxin A (ET A) were detected in 72 isolates by PCR associated genotypic method in this study, none of the isolates carried the genes for enterotoxin D (SED) and exfoliative toxin B (ETB). The occurrence rate of exotoxin genes rated as 12.5%, and the precision of the PCR identification results has been confirmed using the reference strains.

Development of Reverse Transcription Semi-nested PCR Primer Pairs for the Specific and Highly Sensitive Detection of Human Aichivirus A1

  • Lee, Siwon;Cho, Kyu Bong
    • Biomedical Science Letters
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    • v.25 no.4
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    • pp.331-338
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    • 2019
  • Human Aichivirus A1 (HuAiV-A1) is a waterborne human pathogenic virus classified as Picornaviridae and Kobuvirus. In this study, we developed a method that can detect about 35 minutes faster with the same detection sensitivity level than the previously reported HuAiV-A1 diagnostic RT-PCR primer. The RT-PCR primer sets developed in this study are capable of detecting HuAiV-A1 at a level of about 100 ag and formed 563 bp amplification product. In addition, the RT-nested PCR method was able to amplify 410 bp using the RT-PCR product as a template. The detection sensitivity of our method was 10 times higher than the method with the highest detection sensitivity to date. Therefore, the detection method of HuAiV-A1 developed in this study is expected to be used in the water environment in which a small amount of virus exists. Also, this detection method is expected to be used as HuAiV-A1 diagnostic technology in both clinical and non-clinical field.

Comparison of Direct RT-PCR, Cell Culture RT-PCR and Cell IFA for Viability and Infectivity Assay of Cryptosporidium (크립토스포리디움 활성 및 감염성 판정을 위한 direct RT-PCR, cell culture RT-PCR 및 cell culture IFA의 비교)

  • Park, Sang-Jung;Yu, Jae-Ran;Kim, Jong-Min;Rim, Yeon-Taek;Jin, Ing-Nyol;Chung, Hyen-Mi
    • Journal of Life Science
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    • v.16 no.5
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    • pp.729-733
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    • 2006
  • Cryptosporidium is a waterborne pathogenic parasite which causes diarrhea. Immunomagnetic separation-immunofluorescent assay (IMS-IFA) has been a widely adopted for Cryptosporidium detection as standard method. However, this method does not provide information about viability or infectivity of Cryptosporidium. Therefore, many researchers have studied viability or infectivity analyses of Cryptosporidium with various methods such as vital staining, in vitro excystation, RT-PCR, cell culture, and mouse infection assay. In this study, two direct RT-PCR methods, cell culture RT-PCR and cell culture IFA were compared for sensitivity and other characteristics. The results showed that direct RT-PCR method with HSP70 genes had the highest sensitivity with detection up to 1 viable cell of Cryptosporidium. The infectious Cryptosporidium were detected up to 10 to 25 cells by cell culture methods in combination with RT-PCR and IFA. The infectious Cryptosporidium were apt to be quantified by cell culture IFA.

Quantitative Analysis of Feline Calicivirus Inactivation using Real-time RT-PCR (Real-time RT-PCR을 이용한 Feline Calicivirus 불활성화의 정량적 분석)

  • Jeong, Hye Mi;Kim, Kwang Yup
    • Journal of Food Hygiene and Safety
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    • v.29 no.1
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    • pp.31-39
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    • 2014
  • Norovirus causes acute gastroenteritis in all age groups and its food poisoning outbreaks are rapidly increasing in Korea. Reverse transcription-polymerase chain reaction (RT-PCR) is most widely used for the rapid detection of foodborne viruses due to high sensitivity. However, the false positive results of RT-PCR obtained against already inactivated viruses could be a serious drawbacks in food safety area. In this study, we investigated a method to yield true positive RT-PCR results only with alive viruses. To decompose the RNA genes from dead viruses, the enzymatic treatments composed of proteinse K and Ribonuclease A were applied to the sanitized and inactivated virus particles. Another aim of this study was to quantify the efficiencies of several major sanitizing treatments using real-time RT-PCR. Feline calicivirus (FCV) that belongs to the same Caliciviridae family with norovirus was used as a surrogate model for norovirus. The initial level of virus in control suspension was approximately $10^4$ PFU/mL. Most of inactivated viruses treated with the enzymatic treatment for 30 min at $37^{\circ}C$ were not detected in RT-PCR, Quantification results to verify the inactivation efficiencies of sanitizing treatments using real-time RT-PCR showed no false positive in most cases. We could successfully develope a numerical quantification process for the inactivated viruses after major sanitizing treatments using real-time RT-PCR. The results obtained in this study could provide a novel basis of rapid virus quantification in food safety area.

Detection of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus using duplex real-time PCR assay with melting curve analysis on fresh lettuce

  • Lee, Na-Ri;Kwon, Kyung-Yoon;Choi, Sung-Wook;Koo, Min-Seon;Chun, Hyang-Sook
    • Journal of Food Hygiene and Safety
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    • v.26 no.2
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    • pp.114-119
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    • 2011
  • In this study, two duplex real-time PCR approach with melting curve analysis is presented for the detection of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus, which are important food-borne bacterial pathogens usually present in fresh and/or minimally processed vegetables. Reaction conditions were adjusted for the simultaneous amplification and detection of specific fragments in the ${\beta}$-glucuronidase (uidA, E. coli), thermonuclease (nuc, S. aureus), hemolycin (hly, L. monocytogenes) and tetrathionate reductase (ttr, Salmonella spp.) genes. Melting curve analysis using a SYBR Green I real-time PCR approach showed characteristic $T_m$ values demonstrating the specific and efficient amplification of the four pathogens; $80.6{\pm}0.9^{\circ}C$, $86.9{\pm}0.5^{\circ}C$, $80.4{\pm}0.6^{\circ}C$ and $88.1{\pm}0.11^{\circ}C$ for S. aureus, E. coli O157:H7, L. monocytogenes and Salmonella spp., respectively. For all the pathogens, the two duplex, real-time PCR was equally sensitive to uniplex real-time PCR, using same amounts of purified DNA, and allowed detection of 10 genome equivalents. When our established duplex real-time PCR assay was applied to artificially inoculated fresh lettuce, the detection limit was $10^3$ CFU/g for each of these pathogens without enrichment. The results from this study showed that the developed duplex real-time PCR with melting curve analysis is promising as a rapid and cost-effective test method for improving food safety.

Expression of Sodium-Iodide Symporter (NIS) in Thyroid Nodules: Comparison of RT-PCR and Immunohistochemical Staining Methods (갑상선 결절에서 Sodium Iodide Symporter (NIS)의 발현: RT-PCR방법과 면역조직화학염색법의 비교)

  • Bae, Sang-Kyun;Lee, Kang-Dae;Chang, Hee-Kyung
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.6
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    • pp.511-515
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    • 2004
  • Purpose: The sodium-iodide symporter (NIS) expression is an important factor in determining the sensitivity of radioiodine therapy in well-differentiated thyroid cancers. Several previous studies for the expression of NIS in thyroid tissues show diverse results. To investigate whether there is difference between methods in determining the expression of NIS in thyroid tissues of patients with thyroid nodules, we measured the expression ot NIS using two different methods (RT-PCR and immunoshistochemical staining) and compared the results. Materials & Methods: We measured the expression of NIS by reverse transcriptase-polymerase chain reaction (RT-PCR) and also by immunohistochemical staining using anti-NIS antibody in thyroid cancers and other benign thyroid diseases. We compared the results of each method. We included 19 papillary carcinomas, 1 follicular carcinoma, 7 medullary carcinoma, 4 adenomas and 7 nodular hyperplasias. Results: By RT-PCR analysis, 10 of 19 papillary carcinomas expressed NIS, but 1 follicular cancer didn't express NIS. By immunohistochemical staining, 15 of 19 papaillary carcinomas express NIS, but 1 follicular lancer didn't express NIS. There was a significant correlation between the semiquautitative results of RT-PCR and immunohistochemical staining of NIS expression. (p<0.01) Conclusion: Our data demonstrated that the expression of NIS in thyroid cancers and other benign diseases investigated by RT-PCR and immunohistochemical staining correlated well each other. However, by immunohistochemical staining, more NIS expression was found.