• 대한기계학회논문집A
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• 제32권4호
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• pp.371-377
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• 2008

Our precedent study has reported glass-PDMS (polydimethylsiloxane) based biochip for the gene PCR (polymerase chain reaction). To prevent the contamination of bio sample, the once used biochip must not be used repeatedly. However, the fabrication cost of microheater and microsensor of the biochip was not cheap to use it as a disposable chip. This paper proposes new PCR-chip where the glass substrate integrated with the microheater and microsensor is detachable from the reaction chamber where the sample is injected. That makes it possible to reuse the glass substrate repeatedly. The performance of the proposed detachable PCR-chip was compared with that of the precedent monolithic PCR-chip. The results showed that the SRY (sex determining Y chromosome) gene PCR was successfully performed in the detachable chip compared with the monolithic chip. However, the more efforts to improve the efficiency of surface treatment of PDMS chip are needed to increase the possibility of applying the detachable chip to the detecting of male infertility.

• Food Science and Biotechnology
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• 제17권3호
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• pp.569-572
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• 2008

A multiplex polymerase chain reaction (PCR) method was developed to simultaneously detect 3 events of genetically modified (GM) maize. The event-specific primers were used to discriminate the following 3 events of GM maize (DAS-59122-7, TC6275, and MIR604) using multiplex PCR method. The zein gene was used as an endogenous maize reference gene in the multiplex PCR detection. The primer pair Zein-FIR producing a 99 bp amplicon was used to amplify the zein gene. The primer JI-Das-F1/R1 for DAS-59122-7, JI-TC6275-F3/R3 for TC6275, and JI-MIR F1/R1 for MIR604 yielded an amplicon of 130, 162, and 197 bp, respectively. The detection limit of multiplex PCR was 1% for DAS-59122-7, TC6275, and MIR604 for one reaction.

• 대한수의학회지
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• 제38권3호
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• pp.565-573
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• 1998

To study the specific tools for the diagnosis of Newcastle disease virus (NDV) in chicken, polymerase chain reaction (PCR) and its presumable conditions were evaluated for the detection of hemagglutinin-neuraminidase (HN) gene of NDV RNA. For these purposes, Kyojeongwon strain of the NDV was propagated in allantoic cavity of SPF embryonating chicken eggs, and viral RNA was extracted from fractionated virus after the allantoic fluids were ultracentrifuged with sucrose gradient. The first-strand cDNA was then made for the HN gene of NDV RNA by reverse transcription at $42^{\circ}C$ for 1 hour using specific primer complementary to the HN gene. The single-stranded cDNA was used as template in the PCR of the HN-DNA, and various conditions of the PCR were evaluated to set up method for the specific detection of the HN-DNA. The PCR conditions promising for the detection of HN gene consist of preheating at $94^{\circ}C$, 5 min, 30 cycles of denaturation at $94^{\circ}C$, 1 min, annealing at $55^{\circ}C$, 1 min and polymerization at $72^{\circ}C$, 2 min, and a cycle of extension at $72^{\circ}C$, 5 min. when NDVs of allantoic fluids without fractionation were applied to the above PCR condition, the HN genes were detected effectively not only from Kyojeongwon but from other velogenic strains such as Herts and a field isolate.

• 미생물학회지
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• 제38권4호
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• pp.306-311
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• 2002

Methicillin-resistant staphylococci를 신속하게 검출하기 위한 방법으로 mecA polymerase chain reaction을 이용하고 결과를 항생제 감수성 검사와 비교하였다. 국내 병원에 입원중인 환자와 종사자로부터 총 43개의 staphylococci를 분리하여 항생제 감수성 시험을 실시한 결과, 43균주 중 methicillin 내성 균주(MRS)는 39 균주, 감수성 균주(MSS)는 4균주이었다. PCR시험결과, methicillin 내성 균주에서는 MRSC (methicillin-resistant Staphylococcus cohnii), 1 균주(HRC2-4)를 제외하고 mecA 유전자에 해당하는 533bp의 절편이 중폭되었다. 한편, methicillin 감수성으로 분류된 MSSA (methicillin-susceptable Staphylococcus aureus), 1 균주(HSA1-10)에서도 mecA 유전자가 중폭되어 항생제 감수성과 상반된 결과를 보였다. 모든 MSSA는 mecA 양성을나타내었으나, 전체적으로 mecA 유전자의 유무는 항생제 감수성 검사와 95.6%의 일치를 나타냈다.

• 한국임상수의학회지
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• 제27권1호
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• pp.23-28
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• 2010

Mycobacterium avium ssp paratuberculosis, intracellular bacteria that can cause chronic granulomatous enteritis in cattle, continues to pose significant economic losses and health problem with high prevalence. The purpose of this study is the polymerase chain reaction (PCR)-base strategy for early detection of M. avium ssp paratuberculosis in whole blood. Blood samples were collected from korean cattles in Jeonbuk, Korea. The 16 out of 88 serum samples were detected M. partuberculosis by ELISA. Then samples of infected 8 Korean cattles were amplified by PCR. The PCR amplified targets are 16s rDNA and heat shock protein 65kDa (hsp 65). The 16s rDNA provided a highly sensitive and specific tool for the direct detection of mycobacteria. In addition M. avium was confirmed characteristically by the hsp65. Finally there were sure to M. avium ssp paratuberculosis by IS900 PCR. The restriction fragment length polymorphism was identified by PCR amplifications and subsequence restriction enzyme digestions with Pst I of a hsp65. These results indicate that confirm M. avium with 16s rDNA, hsp65 and a restriction fragment length polymorphism in the hsp65 gene can be seem the other pattern. Therefore, these results can be used for clinical direct detections of M. avium ssp paratuberculosis in whole blood of Korean cattle and also to be used epidemiological researches.

• Preventive Nutrition and Food Science
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• 제9권3호
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• pp.276-282
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• 2004

Marginal Zn deficiency is prevalent through the world and yet human zinc status has not been properly assessed due to the lack of a reliable diagnostic indicator. One potential possibility for zinc status assessment using Zn-binding protein, metallothionein (MT)-mRNA, has been proposed. The purpose of the present study was aimed to show whether measurement of mononuclear cell (MNC) MT mRNA, using a competitive-reverse transcriptase-polymerase chain reaction (competitive-RT-PCR) assay, could indicate zinc status in human subjects. In this study, MNC MT-mRNA expression was measured using a competitive-RT-PCR to compare before and after 14 days of zinc supplementation (50 mg Zn/das zinc gluconate). RT-PCR oligonucleotide primers which were designed to amplify both a 278 bp segment of the human MT-2A cDNA and a 198 bp mutant competitor cDNA template from MNCs, were prepared. MT-2A mRNA was normalized by reference to the housekeeping gene, $\beta$-actin, mRNA for which was also measured by competitive-RT-PCR. There was considerable inter-individual variation in MT-mRNA concentration and yet, the mean MT-2A mRNA level increased 4.7-fold after Zn supplementation, as compared to before Zn supplementation. This MT-2A mRNA level was shown as the same pattern and, even more sensitive assay, compared to the conventional plasma and red blood cells (RBCs) Zn assessment in which plasma and RBCs zinc levels increased 2.3- and 1.2-fold, respectively (p<0.05). We suggest that MT competitive-RT-PCR can be a useful assessment tool for evaluating human zinc status.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.592-595
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• 2000

In human chromosome, a short sequence of DNA has been repeated a number of times. These repeats are called variable number of tandem repeat(VNTR) or short tandem repeat(STR) which has short repeat core. VNTR and STR are used in the field of forensic science, evolution, and anthropology. In this work, we examined allele frequencies of 3 VNTR(YNZ22, NeuR, D21S11) and one STR(Humth01) in a Korean population sample by polymerase chain reaction(PCR) followed by high-resolution polyacrylamide gelelectrophoresis(PAGE) with silver staining. Subsequently, the polymorphism information content(PIC) was calculated : the highest PIC was observed for the NeuR locus(0.95680) and lowest for the Humth01 locus(0.75809).

• Journal of Microbiology and Biotechnology
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• 제4권1호
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• pp.20-23
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• 1994

The polymerase chain reaction was used to study the genes inducible under stress from the heavy metal cadmium. Schizosaccharomyces pombe, grown in the presence or absence of sublethal concentration of cadmium, was isolated to purify the total RNAs. The Induced RNA Random Fishing (IRRF) method in which random oligonucleotides were used as primers was applied to the identification of cadmium-induced gene expressions. A PCR-DNA product of 400-bp was cloned and sequenced. Computer analysis showed that this DNA has no homology with any known DNA sequences in GenBank or EMBL databases. The induction of this gene was confirmed by Northern blot analysis of total RNAs isolated from both cadmium-treated and untreated yeast cells.

• Parasites, Hosts and Diseases
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• 제41권2호
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• pp.125-127
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• 2003

Muscle larvae of Trichinella isolates from two outbreaks in Korea were analyzed by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and multiple-PCR. All of the muscle larvae showed a band similar to that of T. spiralis larvae of the reference strain. The two Korean Trichinella isolates (isolate code ISS623 and ISS1078) might be classifiable to Trichinella spiralis.

• 한국어병학회지
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• 제13권1호
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• pp.15-20
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• 2000

자연산 모하의 White Spot Baculovirus(WSBV)보균을 Polymerase Chain Reaction(PCR)에 의한 검출에서 Penaeus chinensis 모하가 52%, Penaeus japonicus 모하는 20%나타났다. 새우 양식장의 서식생물의 본 바이러스의 보균을 조사한 결과 Upogebia major 37%, Calliamassa japonica 29%를 보였다. 본 바이러스의 해수 생존시험에서는 $4^{\circ}C$에서는 장기간 생존하였고 $25^{\circ}C$에서는 시간이 지남에 따라 점차 활성을 잃었다.