• 제목/요약/키워드: reverse transcriptase-polymerase chain reaction (RT-PCR)

검색결과 252건 처리시간 0.025초

C형 간염의 진단을 위한 역전사중합효소연쇄반응과 효소면역측정법의 타당성 평가 (Assessment of Validity of RT-PCR and EIA for The Detection of Hepatitis C Virus Infection)

  • 손병철;전진호;박영홍;신해림;조규일;김종한;정귀옥;이종태;이채언;백낙환
    • Journal of Preventive Medicine and Public Health
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    • 제28권2호
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    • pp.526-541
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    • 1995
  • This study was conducted to estimate the validity of reverse transcriptase-polymerase chain reaction(RT-PCR) compared to enzyme immunoassay(EIA) for the detection of hepatitis C virus (HCV) infection. EIA for antibody to HCV(anti-HCV) and RT-PCR for HCV was executed on the subjects from Pusan and Kyungnam area with questionnaire survey to collect some relating factors of HCV infection. As the result from 617 cases, the prevalence of HCV infection was 1.5% by EIA and 3.7% by RT-PCR(p<0.05), and the age standardized rate was 1.7% and 3.4% by EIA and RT-PCR, respectively. The prevalence of hepatitis B surface antigen(HBsAg) was 6.8% by enzyme linked immunosorbent assay(ELISA) and the age standardized rate was 7.7%. It was the higher in male group comparing to female group(p<0.01). Both of the prevalence of HCV and HBsAg were higher in elevated asparate aminotransferase(AST) and alanine aminotransferase (ALT) group than in normal AST and ALT group(p<0.01). There was no specific risk factor of HCV infection. Though the degree of agreement of EIA and RT-PCR by gamma statistics was 97.2%, it showed a significant difference between the two methods(p<0.01). For the detection of HCV infection, positive predictive value of EIA was 66.7% and negative predictive value of EIA was 97.2%. This study suggests that negative result to anti-HCV by EIA didn't mean the free state of HCV infection, therefore it would be helpful that further monitoring for HCV infection by RT-PCR in the case of elevated AST and ALT and/or clinically suspected.

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SURFACE MICROGROOVES OF THIRTY MICROMETERS IN WIDTH ON TITANIUM SUBSTRATA ENHANCE PROLIFERATION AND ALTER GENE EXPRESSION OF CULTURED HUMAN GINGIVAL FIBROBLASTS

  • Lee, Suk-Won;Kim, Su-Yeon;Lee, Keun-Woo
    • 대한치과보철학회지
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    • 제45권6호
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    • pp.787-794
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    • 2007
  • Statement of problem. Surface microgrooves on Ti substrata have been shown to alter the expression of genes responsible for various biological activities of cultured fibroblasts. However, their effect on enhancing cell proliferation is not yet clear. Purpose. The purpose of this study was to determine the dimension of surface microgrooves on Ti substrata that enhances proliferation and alters gene expression of cultured human gingival fibroblasts. Material and methods. Commercially pure Ti discs with surface microgrooves of monotonous $3.5{\mu}m$ in depth and respective 15 and $30{\mu}m$ in width were fabricated using photolithography and used as the culture substrata in the two experimental groups in this study (TiD15 and TiD30), whereas the smooth Ti was used as the control substrata (smooth Ti group). Human gingival fibroblasts were cultured on the three groups of titanium substrata and the proliferation, DNA synthesis, and gene expression of theses cells were analyzed and compared between all groups using XTT assay, BrdU assay, and reverse transcriptase-polymerase chain reaction (RT-PCR), respectively. Results. From the XTT assay at 48 h incubation, the proliferation of human gingival fibroblasts in TiD30 was significantly enhanced compared to that in smooth Ti and TiD15. The results from the BrdU assay showed that, at 24 h incubation, the DNA synthesis was significantly enhanced in TiD30 compared to that in smooth Ti. In RT-PCR, increase in the expression of PCR transcripts of fibronectin, CDK6, $p21^{cip1}$ genes was noted at 48h incubation. Conclusion. Surface microgrooves $30{\mu}m$ in width and $3.5{\mu}m$ in depth on Ti substrata enhance proliferation and alter gene expression of cultured human gingival fibroblasts.

남해안 자연산 어류에서 Marine birnavirus(MABV)의 검출 (Detection of Marine Birnavirus(MABV) from Marine Fish in the Southern Coast of Korea)

  • 윤현미;김석렬;이월라;정성주;오명주
    • 한국양식학회지
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    • 제21권1호
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    • pp.13-18
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    • 2008
  • Marine birnavirus(MABV)는 중요한 어류 병원체로서 일본 양식산 방어서 처음 분리되어 이후 다양한 해산어종에서 MABV 분리가 보고되었으며 환경적인 샘플인 저질, 해양의 동물성 플랑크톤과 해수에서도 MABV 유전자가 검출되고 있다. 본 연구는 Marine birnavirus disease 에 대한 예방학적 접근의 일환으로서 자연산 어류로부터의 MABV 검출 및 지리적 분포, 보균 어종에 대한 유전학적인 조사를 목적으로 2003년과 2005년도에 동중국해역 3지점, 서해안 5지점 그리고 남해안 5지점에서 어류를 채집하였다. RT-PCR(Reverse transcriptase-Polymerase Chain Reaction)을 이용한 연구결과 자연산 해산어류 160마리 중 13종 17마리에서 MABV가 거출되어 감염율10.6%를 보였다. 조사된 13종의 어류 중 특히 농어목에서 가장 높는 검출율을 보였다. 자연산 어류에서 분리한 MABV 분리균주는 염기서열 분석에서 94.7%-100%, 아미노산 분석에서 97.2-100% 유사성을 나타내었으며 IPNV strain과는 구분되며 MABV에 속하였다.

Cloning and expression of lin-28 homolog B gene in the onset of puberty in Duolang sheep

  • Xing, Feng;Zhang, Chaoyang;Kong, Zhengquan
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권1호
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    • pp.23-30
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    • 2019
  • Objective: Recent studies have demonstrated that lin-28 homolog B (LIN28B)/miRNA let-7 (let-7) plays a role in the regulation of pubertal onset in mammals. However, the role of LIN28B/let-7 in the onset of ovine puberty remains unknown. We cloned the Duolang sheep Lin28B cDNA sequence, detected the expression change of LIN28B, let-7a and let-7g in hypothalamus, pituitary and ovary tissues at three different pubertal stages. Methods: The reverse transcriptase polymerase chain reaction (RT-PCR) was used to clone the cDNA sequence of LIN28B gene from Duolang sheep and the bioinformatics methods were applied to analyze the amino acid sequence of LIN28B protein. The mRNA expression levels of the LIN28B gene at different pubertal stages were examined by real time RT-PCR. Results: LIN28B cDNA of Duolang sheep was cloned, and two transcripts were obtained. The amino acid sequence of transcript 1 shares 99.60%, 98.78%, and 94.80% identity with those of goat, wild yak and pig, respectively. Strong LIN28B mRNA expression was detected in the hypothalamus, pituitary, ovary, oviduct and uterus, while moderate expression was found in the liver, kidney, spleen and heart, weak expression was observed in the heart. No expression was found in the lungs. Quantitative real-time PCR (QPCR) and western-blot analysis revealed that the LIN28B was highly expressed in the hypothalamus and ovary at prepuberty stages, and this expression significantly decreased from the prepuberty to puberty stages (p<0.05). Markedly increased levels of mRNA expression were detected in the pituitary from prepuberty to puberty (p<0.05) and then significantly decreased from puberty to post-puberty (p<0.05). The expression levels of let-7a and let-7g showed no significant changes among different pubertal stages (p>0.05). Conclusion: These results provided a foundation for determining the functions of LIN28B/let-7 and their role in the onset of sheep puberty.

대기행렬을 이용한 위음성률이 있는 코로나 취합검사 시스템의 분석 (The Analysis of COVID-19 Pooled-Testing Systems with False Negatives Using a Queueing Model)

  • 김길환
    • 산업경영시스템학회지
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    • 제44권4호
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    • pp.154-168
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    • 2021
  • COVID-19 has been spreading all around the world, and threatening global health. In this situation, identifying and isolating infected individuals rapidly has been one of the most important measures to contain the epidemic. However, the standard diagnosis procedure with RT-PCR (Reverse Transcriptase Polymerase Chain Reaction) is costly and time-consuming. For this reason, pooled testing for COVID-19 has been proposed from the early stage of the COVID-19 pandemic to reduce the cost and time of identifying the COVID-19 infection. For pooled testing, how many samples are tested in group is the most significant factor to the performance of the test system. When the arrivals of test requirements and the test time are stochastic, batch-service queueing models have been utilized for the analysis of pooled-testing systems. However, most of them do not consider the false-negative test results of pooled testing in their performance analysis. For the COVID-19 RT-PCR test, there is a small but certain possibility of false-negative test results, and the group-test size affects not only the time and cost of pooled testing, but also the false-negative rate of pooled testing, which is a significant concern to public health authorities. In this study, we analyze the performance of COVID-19 pooled-testing systems with false-negative test results. To do this, we first formulate the COVID-19 pooled-testing systems with false negatives as a batch-service queuing model, and then obtain the performance measures such as the expected number of test requirements in the system, the expected number of RP-PCR tests for a test sample, the false-negative group-test rate, and the total cost per unit time, using the queueing analysis. We also present a numerical example to demonstrate the applicability of our analysis, and draw a couple of implications for COVID-19 pooled testing.

인천지역에서 발생한 돼지콜레라의 역학적 특성 (Characteristics of outbreak for the classical swine fever (CSF) at Incheon metropolitan area in 2002)

  • 권효정;변재원;이정구;김경호;박은정;이성모;황현순
    • 한국동물위생학회지
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    • 제26권1호
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    • pp.1-9
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    • 2003
  • The aim of this survey was to investigate the characteristics of outbreak farm determined as the classical swine fever(CSF) at Gangwha-gun and Seo-gu, Incheon metropolitan area from October 7 to November 25 in 2002. Sixty pigs in six different farms were confirmed to the CSF and a total of 9,106 pigs containing 3,194 related epidemiologically was slaughtered to stop spreading of the disease. Clinical signs of pigs diagnosed with the CSF were high fever, anorexia, depression, paralysis of hindlimbs, cyanosis, etc and gross lesions were typically represented with hemorrhage of submandibular and superficial lymph node, infarction of spleen, and petechial (ecchymotic) hemorrhage of kidney and skin. But some outbreak farms had not shown remarkable symptoms, so they were confused with other bacterial diseases. White blood cell (WBC) counts, the classical swine fever virus(CSFV) antigen and antibody enzyme linked immunosorbent assay(ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR) results about six farms indicated that total 60 pigs were infected with the CSFV. Although the origin and infection route of the CSFV were not clear, but the transmissions between farms were mainly through indirect contact such as the movement of farm personal and vehicles from outbreak farm.

Analysis of gene expression during odontogenic differentiation of cultured human dental pulp cells

  • Seo, Min-Seock;Hwang, Kyung-Gyun;Kim, Hyong-Bum;Baek, Seung-Ho
    • Restorative Dentistry and Endodontics
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    • 제37권3호
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    • pp.142-148
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    • 2012
  • Objectives: We analyzed gene-expression profiles after 14 day odontogenic induction of human dental pulp cells (DPCs) using a DNA microarray and sought candidate genes possibly associated with mineralization. Materials and Methods: Induced human dental pulp cells were obtained by culturing DPCs in odontogenic induction medium (OM) for 14 day. Cells exposed to normal culture medium were used as controls. Total RNA was extracted from cells and analyzed by microarray analysis and the key results were confirmed selectively by reverse-transcriptase polymerase chain reaction (RT-PCR). We also performed a gene set enrichment analysis (GSEA) of the microarray data. Results: Six hundred and five genes among the 47,320 probes on the BeadChip differed by a factor of more than two-fold in the induced cells. Of these, 217 genes were upregulated, and 388 were down-regulated. GSEA revealed that in the induced cells, genes implicated in Apoptosis and Signaling by wingless MMTV integration (Wnt) were significantly upregulated. Conclusions: Genes implicated in Apoptosis and Signaling by Wnt are highly connected to the differentiation of dental pulp cells into odontoblast.

Beauvericin, a cyclic peptide, inhibits inflammatory responses in macrophages by inhibiting the NF-κB pathway

  • Yoo, Sulgi;Kim, Mi-Yeon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • 제21권4호
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    • pp.449-456
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    • 2017
  • Beauvericin (BEA), a cyclic hexadepsipeptide produced by the fungus Beauveria bassiana, is known to have anti-cancer, anti-inflammatory, and anti-microbial actions. However, how BEA suppresses macrophage-induced inflammatory responses has not been fully elucidated. In this study, we explored the anti-inflammatory properties of BEA and the underlying molecular mechanisms using lipopolysaccharide (LPS)-treated macrophage-like RAW264.7 cells. Levels of nitric oxide (NO), mRNA levels of transcription factors and the inflammatory genes inducible NO synthase (iNOS) and interleukin (IL)-1, and protein levels of activated intracellular signaling molecules were determined by Griess assay, semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), luciferase reporter gene assay, and immunoblotting analysis. BEA dose-dependently blocked the production of NO in LPS-treated RAW264.7 cells without inducing cell cytotoxicity. BEA also prevented LPS-triggered morphological changes. This compound significantly inhibited nuclear translocation of the $NF-{\kappa}B$ subunits p65 and p50. Luciferase reporter gene assays demonstrated that BEA suppresses MyD88-dependent NF-${\kappa}B$ activation. By analyzing upstream signaling events for $NF-{\kappa}B$ activation and overexpressing Src and Syk, these two enzymes were revealed to be targets of BEA. Together, these results suggest that BEA suppresses $NF-{\kappa}B$-dependent inflammatory responses by suppressing both Src and Syk.

Expression of DSPP mRNA During Differentiation of Human Dental Pulp-derived Cells (HDPC) and Transplantation of HDPC Using Alginate Scaffold

  • Aikawa, Fumiko;Nakatsuka, Michiko;Kumabe, Shunji;Jue, Seong-Suk;Hayashi, Hiroyuki;Shin, Je-Won;Iwai, Yasutomo
    • International Journal of Oral Biology
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    • 제31권3호
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    • pp.73-79
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    • 2006
  • Tissue stem cells are used for the regenerative medicine. In previous study we observed hard tissue formation of human dental pulp-derived cells using alginate scaffold. In this study, we explore the ability to differentiate of the 13th passage cells with glycerol 2-phosphate disodium salt hydrate (${\beta}-GP$) which accelerate calcification. Reverse transcriptase Polymerase Chain Reaction (RT-PCR), transplants using alginate scaffold and histological examination were performed. We observed the expression of DSPP mRNA on day 10 cultured cells with ${\beta}-GP$. In conclusion, the 13th passage cells still have an ability to differentiate into odontoblast-like cells and alginate supports the differentiation of cultured cells in the transplants.

계지복령환이 Cholesterol 식이 생쥐의 SOD 유전자발현능에 미치는 영향 (Effects of geagibokrounghwan on superoxide dismutase gene expression in experimental mouse fed cholesterol)

  • 홍윤표;노승현
    • 대한한의학방제학회지
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    • 제6권1호
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    • pp.215-226
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    • 1998
  • Geagibokrounghwan (桂技茯笭湯) has long been used to cure human diseases such as vascular and blood disorders. However, it is still unkown on its action mechanism, physiolosical and biochemical meaning. Thus, many attempts were tried to show the scientific background covering the above mentioned mechanism. The effect of Geagibokrounghwan, which was known to date, as follow; effective circulation of body blood system, proliferation of leucocytes and antioxidative action. In this study, we have applied the Geagibokrounghwan administration and feed to mouse, to see effects on the expression of superoxide dismutase(SOD) mRNA as antioxidative agent and oxygen radical scavenger. Total RNAs includingmRNA have been isolated from liver and white blood cells after mice were fed with cholesterol in high dose. Also, in a separate group, the cholesterol-administrated mice were fed with Geagibokrounghwan to see the effects on SOD transcription. and then reverse transcriptase-polymerase chain reaction (RT-PCR) usion each primer set (SOD-F;GATGAAAGCGGTGT-3'; SOD-R; 5'-CCTGTGGAGTGATT-3') were performed to trace theamounts of mRNA. SOD mRNA was specifically expressed in Geagibokrounghwan-fed mice at 2 weeks after treatment, however, gradually reduced after 4 weeks. These results indicate that Geagibokrounghwan is highly applicable in treatment of the above mentioned human diseases.

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