• Title/Summary/Keyword: C&RT

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Specific Primer Sets for RT-PCR Detection of Major RNA Viruses of Tomato Plants in Korea (국내의 토마토 주요 바이러스 진단을 위한 역전사중합반응법용 프라이머 세트)

  • Shin, Jun-Sung;Han, Jung-Heon;Shin, Yu-Ju;Kwak, Hae-Ryun;Choi, Hong-Soo;Kim, Jeong-Soo
    • Research in Plant Disease
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    • v.23 no.2
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    • pp.193-201
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    • 2017
  • Major tomato viruses in Korea are Tomato chlorosis virus (ToCV), Tomato spotted wilt virus (TSWV), Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), and Tomato mosaic virus (ToMV). RT-PCR conditions for the viruses were examined, especially in primer set and RT-PCR mixture. Total 46 primer sets from the unique sequence of the viruses were tested for nonspecific background products in a RT-PCR mixture without template. Among them 16 primer sets were applied to healthy tomato RNA, resulting the compatibility between RT-PCR mixture and primer set influenced RT-PCR to reduce nonspecific background products. Based on the combinations among cDNA synthesis parameters and RT-PCR mixtures, two reaction mixtures were finally selected for ToCV detection. The condition allowed to determine more specific primer sets; C029 (ToCV), C072 (TSWV), C070 (CMV), C048 (PepMoV), and C065 (ToMV). These primer sets are expected to be of use to specific detection of the major viruses in tomato plants.

Comparison of the Pathogenicity of Infectious Hematopoietic Necrosis Virus Genotypes Isolated from Rainbow Trout in Gangwon Province (무지개송어에서 분리된 IHNV (감염성 조혈 괴사바이러스) 유전자형에 따른 병원성 비교)

  • Lee, Chang-Ju;Kim, Kwang-Il;Han, Yu-Seon;Jegal, Myeong-Eun;Kim, Yung-Jin
    • Journal of Life Science
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    • v.31 no.6
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    • pp.574-580
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    • 2021
  • This study investigated the pathogenicity of different genotypes of infectious hematopoietic necrosis virus (IHNV) strains isolated from infected rainbow trout in Gangwon Province. RtPc0314c and RtPc0314g strains belonging to the JRt-Shizuoka lineage and RtPc0816g strain belonging to the JRt-Nagano lineage showed 100% cumulative mortality when inoculated at a high titer. In addition, more rapid necrosis was observed in rainbow trout infected with RtPc0314c and RtPc0314g mutations. When inoculated at a low titer, 100% mortality was not observed until the end of the experiment, but the mortality was higher in rainbow trout infected with a mutant strain belonging to the JRt-Shizuoka linage than a mutant strain belonging to the JRt-Nagano lineage. A histopathological analysis showed a clear signature of infection in kidney and spleen tissues upon infection with RtPc0314c and RtPc0314g but no signature of infection associated with the Rt03186 strain. Based on the results in this study, it seems that strains belonging to the JRt-Shizuoka lineage in Gangwon Province IHNV are more pathogenic.

Bioavailability of Ranitidine Tablets in Rats (흰쥐에서 라니티딘제제의 생체이용률)

  • 이미숙;구영순
    • YAKHAK HOEJI
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    • v.39 no.6
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    • pp.636-644
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    • 1995
  • Comparison of bioavailabflity (BA) of three brands of ranitidine (RT) tablets has been studied m rats. The purpose of this study was to characterize the pharniacolunetics of RT tablets in the rat and to coinpare phannacolunetic parameters of three brands of RT tablets. In addition, it was investigated whether plasma RT concentrations m humans can be predicted from pharmacokinetic parameters obtained in rats. RT was administered intravenously in dose of RT.HCI 10mg/kg and orally in dose of RT.HCI 50mg/kg as solution or crushed sample of thablets. Plasma RT concentrations were determned by HPLC. Plasma RT concentrations as a function of time were fitted to two compartment model. Plasma RT concentrations declined with a terminal half life ($t_{{1}/2{\betha}}$) of 40.9 min. The plasma RT concentration-time curve showed two peak plasma concentrations following an oral administration of solution or crushed sample in rats like humans. No significant difference among pharmacokinetic parameters was observed except $T_{max2}$ (p<0.05). The BA for crushed sample A, B and C were found to be 54.6 40.7 and 40.0%, respectively. Equivalence of $C_{max1}$ and $T_{max2}$ were guaranteed in this study. However, it was concluded that three brands of RT tablets are bioequivalent, taking the following characteristics of RT into consideration;(1) rapid onset of the effect is not required, (2) $C_{max1}$ and $T_{max2}$ do not seem to influence the effectiveness of the drug during a long-term treatment by the usual administration of twice a day. Results from this study were combined with plarmacokinetic data for RT in dogs and humans to develop a basis for interspecies scale-up of the disposition characteristics of the drug. there were similarities in the general disposition of the drug. Allometric relationships were sought between pharmacokinetic parameters nd species body weight. Significant interspecies correlations were found for total body clearance($Cl_{t}$) and steady state volume of distribution ($Bd_{ss}$). Thus, plasma RT concentrations in humans can be predicted from pharmacokinetic parameters obtained in rats.

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Efficient Diagnosis of Cucumber Green Mottle Mosaic Virus in Watermelon Using RT-PCR and Cloning of Coat Protein Gene (RT-PCR을 이용한 수박 Cucumber Green Mottle Mosaic Virus의 효율적인 진단 및 외피단백질 유전자의 클로닝)

  • 양덕춘;이진숙;김두욱;임용표;민병훈
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.519-524
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    • 1998
  • A simple and reliable method to diagnose cucumber green mottle mosaic virus of watermelon in Korea (CGMMV-WK) was determined by RT-PCR, and coat protein gene for CGMMV-WK was cloned. Comparing to a method reported by Lee et al. (1996), the method developed here showed a better RT-PCR reaction. RT-PCR was possible by one step in the PCR reaction mixture that contains 20 pmol of primer, reverse transcriptase (30 unit), RNasin (5 unit) using the crude RNA solution. RT-PCR condition for specifically diagnosing CGMMV-WK was that cDNA was synthesized at 42$^{\circ}C$ for 45 min followed by pre-denaturation at 95$^{\circ}C$ for 2 min, and then PCR reaction was carried out with a programmed condition that consisted of 36 sequential cycles at 96$^{\circ}C$ for 30 sec, 6$0^{\circ}C$ for 30 sec, and 72$^{\circ}C$ for 1 min. A gene encoding the coat protein of CGMMV-WK was cloned and characterized. Nucleotide sequence of coat protein gene of CGMMV-WK shared 98.77% and 99.38% of sequence identity with those of CGMMV-W and CGMMV-SH, respecitvely, however, all of amino acid sequences were same.

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The Results of Radiation Therapy Alone vs Radiation Plus Chemotherapy of Uterine Cervix Cancer (자궁경부암에서 방사선 단독요법 및 항암화학과 방사선 병용요법의 치료성적)

  • Lee, Myung-Za;Choi, Seog-Young;Chun, Ha-Chung
    • Radiation Oncology Journal
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    • v.13 no.2
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    • pp.181-189
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    • 1995
  • Pupose: Radiation therapy(RT) is conventionally standard treatment for locally advanced stage for uterine cervix cancer. Recently to improve treatment results, combined chemotherapy and radiation therapy was tried We retrospectively analysed our experience of 122 patients. Comparision of the results in 45 patients treated with RT alone and 77 patients treated with RT plus chemotherapy was made Materials and Mathods: From January 1985 to December 1991 122 patients with cervix cancer were treated with whole pelvic external RT and ICR(34 1 ICR, 77 2 ICR, 11 high dose rate ICR) in our department. Forty five patients were treated with RT alone, and 77 patients were treated with combined RT plus chemotherapy Mean age was 58 years(range:29-81). Histologic types were 111 squamous cell carcinoma, 5 large cell carcinoma, 3 adenocarcinoma, and 2 adenosquamous cell carcinoma. According to the FIGO stage 6 had stage $IA(4.9\%),$ 11 had $IIA(9.0\%),$ 37 had $IIB(30.3\%),$ 3 had $IIIA(2.5\%),$ 63 had $IIIB(51.6\%).$ and 2 had stage $IV(1.6\%).$ In 77 patients with RT Plus chemotherapy, 36 patients were treated with VBP(vinblastin, bleomycin, cisplatinum) , 39 patients with cisplatinum plus 5-FU and 2 patients with 5-FU. Results: Complete response after external RT (3960cGy-5500cGy) was achieved in 61 patients$(50\%).$ The actuarial 5 year and 9 rear survival rate was $57.8\%\;and\;53.9\%,$ respectively. Five rear actuarial survival rate was $63.1\%$with RT alone(n=45) and $55.9\%$ with RT plus chemotherapy(n=77). The 5 rear survival rate was $35.5\%$ for 1 course of ICR and $67\%$ for 2 courses of ICR. There was statistically significant advantage of survival with RT alone group who were treated with 2 courses of ICR and dose to the A Point)=8000cGy (4/25 died). In RT plus chemotherapy group, dose response was not seen and there was no difference in 5 year survival between 1 course and 2 course of ICR $(50\%\;vs\;56.8\%),$ and dose to point A less than 8000 cGy and more than 8000 $cGy(55.6\%\;vs\;55.7\%).$ There was no significant difference in survival between RT alone and RT plus chemotherapy for patients with tumor size greater than 3cm in size. Five year survival rate for early stage (Stage IB and IIA) with RT alone group and with RT Plus chemotherapy group was $60\%\;and\;77.0\%,$ respectively In advanced stage (stage IIB, IIIA, IIIB, IVA) the 5 year actuarial survival rate were $62.6\%,$ for RT alone group vs $53.6\%$ for RT plus chemotherapy group. Conclusion: Present study demonstrates that there is no survival advantage with adding chemotherapy in advanced stage of uterine cervix cancer. RT alone is considered as treatment of choice for patients with locally advanced cervix cancer. There was increased survival in RT alone group treated with RT dose above 8000 cGy to point A and 2 course of ICR. but 2 course of ICR and RT dose above 8000 cGy to point A did not affect survival advantage in RT plus chemotherapy group.

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RT-PCR for detection of group A, B and C porcine rotaviruses (RT-PCR 기법을 이용한 돼지 로타바이러스 주요 혈청법 감별진단)

  • Park, Choi-kyu;Lyoo, Young-soo;Paul, Prem S.
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.85-89
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    • 1998
  • 본 연구에서는 돼지 로타바이러스의 혈청형 A, B 및 C를 동시에 진단할 수 있는 RT-PCR 기법을 개발하였다. 각각의 혈청형에 특이적인 primer를 이용하여 RT-PCR 기법으로 분변시료에서 직접 바이러스 동정을 실시한 결과, 23건의 로타바이러스 감염 분변 시료에서 13건이 혈청형 A, 3건이 혈청형 B, 2건이 혈청형 C로 나타나 국내에서도 A, B 및 C형의 돼지 로타바이러스가 공히 발생하고 있음을 확인하였으며, 발생분포는 외국의 예와 유사하였다. 이 RT-PCR기법은 돼지 로타바이러스의 주요 혈청형인 A, B 및 C형의 동시감별진단법으로 이용될 수 있을 것으로 판단된다.

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A Simple Detection of Sweetpotato Feathery Mottle Virus by Reverse Transcription Polymerase Chain Reaction

  • Jeong Jae-Hun;Chakrabarty Debasis;Kim Young-Seon;Eun Jong-Seon;Choi Yong-Eui;Paek Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.5 no.2
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    • pp.83-86
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    • 2003
  • A reverse transcription polymerase chain reaction (RT-PCR) protocol was developed using two specific 22-mer primers located in coat protein gene of SPFMV. A 411 bp PCR-product was detected in virus infected plants as well as tissue culture raised sweet potato but not in healthy plants. For optimization of RT-PCR protocol, the optimum crude nucleic acid concentration, annealing temperature, primer concentration and numbers of PCR-cycle for maximum sensitivity and specificity were determined. The optimum condition for RT-PCR was as follows: RT-PCR reaction mixture was one-step mixture, containing 50 pmol of primer, 30 units of reverse transcriptase, 5 units of RNasin, and the crude nucleic acid extracts (200 ng). In RT-PCR, cDNA was synthesized at $42^{\circ}C$ for 45 min before a quick incubation on ice after pre-denaturation at $95^{\circ}C$ for 5 min. The PCR reaction was carried out for 40 cycles at $96^{\circ}C$ for 30 see, $63^{\circ}C$ for 30 sec, $72^{\circ}C$ for 1 min, and finally at $72^{\circ}C$ for 10 min. The viral origin of the amplified product was confirmed by sequencing, with the sequence obtained having $95-98\%$ homology with published sequence data for SPFMV. The benefits of this RT-PCR based detection of SPFMV would be simple, rapid and specific.

Rapid and Sensitive Detection of Hepatitis C Virus in Clinical Blood Samples Using Reverse Transcriptase Polymerase Spiral Reaction

  • Sun, Wenying;Du, Ying;Li, Xingku;Du, Bo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.459-468
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    • 2020
  • This study established a new polymerase spiral reaction (PSR) that combines with reverse transcription reactions for HCV detection targeting 5'UTR gene. To avoid cross-contamination of aerosols, an isothermal amplification tube (IAT), as a separate containment control, was used to judge the result. After optimizing the RT-PSR reaction system, its effectiveness and specificity were tested against 15 different virus strains which included 8 that were HCV positive and 7 as non-HCV controls. The results showed that the RT-PSR assay effectively detected all 8 HCV strains, and no false positives were found among the 7 non-HCV strains. The detection limit of our RT-PSR assay is comparable to the real-time RT-PCR, but is more sensitive than the RT-LAMP. The established RT-PSR assay was further evaluated for detection of HCV in clinical blood samples, and the resulting 80.25% detection rate demonstrated better or similar effectiveness compared to the RT-LAMP (79.63%) and real-time RT-PCR (80.25%). Overall, the results showed that the RT-PSR assay offers high specificity and sensitivity for HCV detection with great potential for screening HCV in clinical blood samples.

RT- PCR Analysis of Vitellogenin Gene Expression in Bombina orientalis (무당개구리 비텔로제닌 유전자의 발현의 RT- PCR 검출법)

  • 계명찬;이명식;강희정;정경아;안혜선
    • Korean Journal of Environmental Biology
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    • v.22 no.2
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    • pp.329-335
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    • 2004
  • To develop a biomarker for the monitoring of the contamination of estrogenic endocrine disrupters in the aquatic environment, reverse transcription -polymerase chain reaction (RT-PCR) analysis of vitellogenin (Vg) mRNA expression was optimized in Bombina orientalis, a Korean red bellied toad species. Based on partial cDNA sequences of both Vg and beta actin genes of B. orientalis, specific primers for RT-PCR of Vg and beta actin mRNAs were developed. Semiquantitative RT-PCR of the Vg mRNA in liver was optimized using a beta actin mRNA as an internal control in both sexes. In female RT-PCR using $1\;\mu{g}$ of the liver cDNA resulted in a linear increment in the PCR product of Vg from 18 to 34 cycles of amplification. In male, on the contrary, the RT- PCR product was first detected at 30 cycles of amplification and a linear increment was observed from 30 to 40 cycles of amplification, suggesting that male B. orientalis expresses minute amount of Vg mRNA which is a $2^{-12}$ equivalent of female. In conclusion, the optimized protocol for semiquantitative RT-PCR analysis of Vg mRNA level in B. orientalis male liver will be useful for the environmental monitoring the xenoestrogen contamination in the freshwater environment in Korea.