• Title/Summary/Keyword: reverse-transcription polymerase chain reaction

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Nucleotide Sequence of Hop Stunt Viroid Kh Strain (HSVd-Kh) (호프 왜화바이로이드 Kh 계통 (HSVd-Kh)의 염기서열)

  • 이재열;김경숙;정동수
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.612-613
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    • 1998
  • The nucleotide sequence of hop stunt viroid HHSVd) Kh strain was sequenced by the reverse transcription and polymerase chain reaction. It consists of 296 nucleotides, and differs by one nucleotide deletion of cytosine at the position of 295 from the HSVd-K strain which consists of 297 nucletoides.

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Severe Fever with Thrombocytopenia Syndrome Patients with Hemophagocytic Lymphohistiocytosis Retrospectively Identified in Korea, 2008-2013

  • Kim, Kye-Hyung;Lee, Myung Jin;Ko, Mee Kyung;Lee, Eun Yup;Yi, Jongyoun
    • Journal of Korean Medical Science
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    • v.33 no.50
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    • pp.319.1-319.5
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    • 2018
  • The incidence of severe fever with thrombocytopenia syndrome (SFTS) has increased in Korea since a first report in 2013. We investigated whether SFTS existed before 2013 using real-time reverse transcription polymerase chain reaction and stored blood samples from febrile patients with thrombocytopenia. Four cases of SFTS were identified, with the earliest occurring in 2008.

Determination of Tyrosinase mRNA in Melanoma by Reverse Transcription-PCR and Optical Mirror Resonance Biosensor

  • Taeboo Choe;Park, Inchul;Seokil Hong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.4
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    • pp.212-215
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    • 2002
  • Tyrosinase transcript In the blood Is known as the marker of malignant melanoma and it has been often determined by using reverse transcription-polymerase chain reaction (RT-PCA) . However, after the PCR process, the quantification of amplified CDMA by the gel electrophoresis is not reliable and time-consuming. for this reason, we tried to quantify the PCR product using a cuvette-type biosensor, where the oligonucleotide probe was immobilized on the cuvette surface and the single strand CDMA, the denatured PCH product, was then hybridized onto the immobilized probe to give a response signal. The response was Immediate and takes 15 min to obtain a stable signal. The biosensor was much more sensitive comparing to the gel electrophoresis method. The quantification of PCR product using a cuvette-type biosensor was feasible and rapid.

Quantitation of Hepatitis C Viral RNA Using Direct CRT-PCR

  • Park, Young-Suk;Lee, Kyung-Ok;Oh, Moon-Ju;Chai, Young-Gyu
    • BMB Reports
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    • v.30 no.3
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    • pp.234-236
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    • 1997
  • Chronic hepatitis C virus (HCV) infection is associated with the rapid development of cirrhosis and hepatocellular carcinoma. It has been reported that the amount of HCV RNA may be correlated with the progression of hepatitis and may be a prognostic marker for treatment of HCV patients. The direct detection of HCV RNA by reverse transcription-polymerase chain reaction (RT-PCR) is widely used to determine the presence of circulating virions. The most relevant limit of this approach is the lack of quantitative information about the viral titer. In the present study, we developed the method for HCV quantitation using competitive reverse transcription (CRT)-PCR using the deleted HCV standard. The serially diluted standard was added in titrated amounts to the target HCV RNA. The mixture was then reverse transcribed and amplified in the same reaction tube. The methods were evaluated using over 110 HCV-PCR positive samples in Koreans. About 59% of the samples were judged to contain $10^{5}-10^{6}$ copies of HCV RNA in 1 ml of serum.

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Application of Buoyant Density Centrifugation Method for the Rapid Detection of Feline Calicivirus in Oyster and Lettuce as Norovirus Surrogate

  • Cho, Yun-Sik;Lee, Kang-Whie;Jang, Keum-Il;Ahn, Jun-Bae;Kim, Kwang-Yup
    • Food Science and Biotechnology
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    • v.17 no.5
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    • pp.925-930
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    • 2008
  • Norovirus has become the most common cause of human gastroenteritis in developed countries. Detection procedures of foodborne viruses from foods require several steps. The concentration step using polyethylene glycol (PEG) is time-consuming and the detection efficiency of reverse transcription-polymerase chain reaction (RT-PCR) is affected by inhibitors from food components. In this study, a rapid detection method based on buoyant density centrifugation was developed to replace the time-consuming chloroform-polyethylene glycol-Tris Tween method. Feline calicivirus that belongs to the family Caliciviridae was used as a surrogate model for norovirus. After artificial inoculation of feline calcivirus (FCV) to oyster and lettuce, 830 ${\mu}L$ of homogenized sample suspension was layered on the top of 670 ${\mu}L$ 20% percoll and centrifuged. Then RNA extraction step was proceeded with the supernatant. By varying several physical conditions, the detection limits were lowered to $2.4{\times}10^2$ PFU per 1 g in oyster and $2.4{\times}10^0$ PFU per 1 g in lettuce. The protocol obtained in this study could be used to develop new detection method for norovirus in foods.

Recent viral pathogen in acute gastroenteritis: a retrospective study at a tertiary hospital for 1 year

  • Jin, Hye Il;Lee, Yoo Mi;Choi, You Jin;Jeong, Su Jin
    • Clinical and Experimental Pediatrics
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    • v.59 no.3
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    • pp.120-125
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    • 2016
  • Purpose: Viral gastroenteritis among children is mainly caused by rotavirus, norovirus, astrovirus, or adenovirus strains. However, changing socioeconomic conditions and a rotavirus vaccination program may be affecting the prevalence of these viral infections. Therefore, we aimed to elucidate the season-specific trends in viral infections for facilitating prophylaxis and surveillance in our region. Methods: We evaluated 345 pediatric patients (203 males, 142 females; age, 1 month to 16 years) who visited the CHA Bundang Medical Center because of gastroenteric symptoms between June 2014 and May 2015. The specimens were simultaneously tested for norovirus, rotavirus, astrovirus, and adenovirus via multiplex reverse transcription polymerase chain reaction. Clinical characteristics of patients were analyzed retrospectively. Results: The most common virus was norovirus, followed by rotavirus, adenovirus, and astrovirus. Of all viral infections, 45.2% occurred mainly between 6 and 24 months of age; in particular, norovirus infection mostly occurred in all age groups except those below 6 months of age, when rotavirus was most prevalent. In addition, seasonal variation was observed, such as norovirus infection from December to February, rotavirus infection from February to April, and adenovirus infection from July to October. Conclusion: Our results showed that the most common cause of acute pediatric viral gastroenteritis had changed from rotavirus to norovirus in our patients, because of effective rotaviral vaccination. We recommend the management of food and personal hygiene in accordance with age or seasons as well as active vaccination for preventing viral gastroenteritis.

Bone Healing Capacity of Demineralized Dentin Matrix Materials in a Mini-pig Cranium Defect

  • Kim, Jong-Yub;Kim, Kyung-Wook;Um, In-Woong;Kim, Young-Kyun;Lee, Jeong-Keun
    • Journal of Korean Dental Science
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    • v.5 no.1
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    • pp.21-28
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    • 2012
  • Purpose: In this study the bone healing ability of autogenous tooth bone graft material as a substitute material was evaluated in a mini-pig cranial defect model through histologic examinations and osteonectin reverse transcription polymerase chain reaction (RT-PCR) quantitative analysis. Materials and Methods: A defect was generated in the cranium of mini-pigs and those without a defect were used as controls. In the experimental group, teeth extracted from the mini-pig were manufactured into autogenous tooth bone graft material and grafted to the defect. The mini-pigs were sacrificed at 4, 8, and 12 weeks to histologically evaluate bone healing ability and observe the osteonectin gene expression pattern with RT-PCR. Result: At 4 weeks, the inside of the bur hole showed fibrosis and there was no sign of bone formation in the control group. On the other hand, bone formation surrounding the tooth powder granule was observed at 4 weeks in the experimental group where the bur hole was filled with tooth powder. Osteonectin gene expression; there was nearly no osteonectin expression in the control group while active osteonectin expression was observed from 4 to 12 weeks in the experimental group. Conclusion: We believe this material will show better results when applied in a clinical setting.

Detection of Poliovirus in Water by Cell Culture and PCR Methods (세포배양법과 PCR 방법에 의한 물에서의 폴리오 바이러스 검출)

  • 조연희;이찬희
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.198-204
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    • 2002
  • Poliovirus is a member of enterovirus which causes paralytic poliomyelitis, encephalitis and aseptic meningitis. Since poliovirus is spread by the fecal-oral route and poliovirus-contaminated water could be a potential threat for public health, detection of poliovirus in drinking water resource is important. Infectious poliovirus and poliovirus inactivated by heat or UV were used to test three detection methods such as cell culture method, reverse transcription-polymerase chain reaction (RT-PCR) and integrated cell culture (ICC)-PCR. Infectious poliovirus was detected by all three methods and ICC-PCR was the most sensitive and fast in detecting poliovirus. Inactivated polioviruses could not be detected by cell culture or ICC-PCR methods. On the other hand, heat- inactivated viruses could be detected by RT-PCR. Thus it is suggested that ICC-PCR method is the most sensitive and effective in detecting infectious polioviruses in water sample.

Development and Assessment of Specific and High Sensitivity Reverse Transcription Nested Polymerase Chain Reaction Method for the Detection of Aichivirus A Monitoring in Groundwater (지하수 중 Aichivirus A 모니터링을 위한 특이적 및 고감도 이중 역전사 중합효소연쇄반응 검출법 개발 및 평가)

  • Bae, Kyung Seon;Kim, Jin-Ho;Lee, Siwon;Lee, Jin-Young;You, Kyung-A
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.190-198
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    • 2021
  • Human Aichivirus (Aichivirus A; AiV-A) is a positive-sense single-strand RNA non-enveloped virus that has been detected worldwide in various water environments including sewage, river, surface, and ground over the past decade. To develop a method with excellent sensitivity and specificity for AiV-A diagnosis from water environments such as groundwater, a combination capable of reverse transcription (RT)-nested polymerase chain reaction (PCR) was developed based on existing reported and newly designed primers. A selective method was applied to evaluate domestic drinking groundwater samples. Thus, a procedure was devised to select and subsequently identify RT-nested PCR primer sets that can successfully detect and identify AiV-A from groundwater samples. The findings will contribute to developing a better monitoring system to detect AiV-A contamination in water environments such as groundwater.

A survey of viruses and viroids in astringent persimmon (Diospyros kaki Thunb.) and the development of a one-step multiplex reverse transcription-polymerase chain reaction assay for the identification of pathogens

  • Kwon, Boram;Lee, Hong-Kyu;Yang, Hee-Ji;Kim, So-Yeon;Lee, Da-Som;An, ChanHoon;Kim, Tae-Dong;Park, Chung Youl;Lee, Su-Heon
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.193-206
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    • 2022
  • Astringent persimmon (Diospyros kaki Thunb.) is an important fruit crop in Korea; it possesses significant medicinal potential. However, knowledge regarding the pathogens affecting this crop, particularly, viruses and viroids, is limited. In the present study, reverse transcription-polymerase chain reaction (RT-PCR) and high-throughput transcriptome sequencing (HTS) were used to investigate the viruses and viroids infecting astringent persimmons cultivated in Korea. A one-step multiplex RT-PCR (mRT-PCR) method for the simultaneous detection of the pathogens was developed by designing species-specific primers and selecting the primer pairs via combination and detection limit testing. Seven of the sixteen cultivars tested were found to be infection-free. The RT-PCR and HTS analyses identified two viruses and one viroid in the infected samples (n = 51/100 samples collected from 16 cultivars). The incidence of single infections (n = 39/51) was higher than that of mixed infections (n = 12/51); the infection rate of the Persimmon cryptic virus was the highest (n = 31/39). Comparison of the monoplex and mRT-PCR results using randomly selected samples confirmed the efficiency of mRT-PCR for the identification of pathogens. Collectively, the present study provides useful resources for developing disease-free seedlings; further, the developed mRT-PCR method can be extended to investigate pathogens in other woody plants.