• Journal of Veterinary Clinics
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• v.15 no.1
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• pp.146-150
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• 1998

소 림프구내의 Theileria sergenti감염을 확인하기 위하여 T. sergenti감염혈액에서 림프구를 분리한 후 중합효소연쇄반웅을 실시하였다. 또한,분리한 림프구내의 T. sergenti감 염을 증명하기 위하여 IFA test와 acridine orange stain을 실시하였다. 그 결과 다음과 같은 성적을 얻을 수 있었다. T. sergenti 감염헐액의 전혈과 림프구를 각각 생리식염수로 2배율 연 속회석하여 중합효소연쇄반응을 실시한 결과, 림프구내에서는 1,024배 회석배율까지 T. sergenti의 genomic DNA가 중폭되었으며, 전혈내에서는 256배 회석배율까지 증폭되었다. 그리 고 중합효소연쇄반응으로 T. sergenti 감염이 확인된 림프구를 이용하여 IFA test와 acridine orange 염색을 실시한 결과, 림프구내에 T. sergenti가 존재하는 것을 증명할 수있었다. 한편, 중합효소연쇄반응을 이용한 림프구내의 T. sergenti 감염의 진단 유용성을 확인하기 위하여 전 북지역에서 사육중인 소 16두를 대상으로 이들의 혈액으로 PCR 증폭을 실시하였다. 그 결과 전혈에서 genomic DNA를 취한 경우에는 3두(18.8%)만이, 그리고 림프구에서 genomic DNA를 취한 경우에는 11두(68.8%)의 소에서 T. sergenti DNA의 증폭을 관찰할 수 있었다.

• Tuberculosis and Respiratory Diseases
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• v.45 no.5
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• pp.935-941
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• 1998

Background: As living and dead Mycobacteria could be amplified by polymerase chain reaction(PCR), it was considered that PCR was inappropriate for the monitoring of pulmonary tuberculosis after treatment. But we found negative conversion of PCR after successful treatment. We would like to know about the negative conversion rate of PCR and its conversion time after antituberculous treatment. Methods: We collected 113 sputums from the 16 patients of pulmonary tuberculsosis visiting Catholic University Hospital of Taegu Hyosung. We consecutively tested AFB smear, AFB culture and PCR by 2 to 4 weeks after antituberculous therapy. The patients were classified according to the chest X ray findings. Results: We detected negative conversion of PCR from all 16 patients of the pulmonary tuberculosis within 30 weeks after treatment. The average negative conversion time was $16{\pm}8$ weeks. The conversion time according to the chest X -ray findings were as follows : For the 8 cases of minimum were $9{\pm}5$ weeks, 4 cases of modreate advanced were $20{\pm}8$ weeks, and 4 cases of far advanced were $23{\pm}2$ weeks. The product of PCR was gradually decreased according to the duration of treatment. Conclusions: From the results of our study, we could utilize M. tubercuosis PCR for the prediction of therapy response and monitoring of the patient with pulmonary tuberculosis after treatment.

• Biomedical Science Letters
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• v.2 no.2
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• pp.241-247
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• 1996

Poor yields of amplified DNAs could be resulted in polymerase chain reaction(PCR) processes with G+C-rich DNA primers because of their high $T_m$ values. To maximize the yields of amplification in PCR processes with G+C-rich primers, we compared the yields of amplified DNA fragments according to the concentrations of specific denaturants added to the reaction mixture of PCR system. With addition of the mixture of 2.5% glycerol and 1.25% formamide, or 2.5% dimethyl sulfoxide to the reaction cocktail, respectively, remarkable increases in the yields of amplified DNA fragments were not observed in the PCR systems with G+C-low primers of Lyl chromosomal gene from Borrelia burgdorferi but observed in the PCR system with G+C- ich primers of Is900 gene from Mycobacterium parahberculosis. Although we were not practically able to discriminate the yields of PCR DNAs according to the concentrations used in this study, addition of the mixture of 5% glycerol and 2.5% formamide, or 5% DMSO tended to increase the production of extra bands.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.2
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• pp.88-93
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• 2016

Dirofilaria immitis is a filarial nematode parasite that causes cardiopulmonary dirofilariasis in dogs. The purpose of this study was the development of real-time PCR assays for efficient detection of D. immitis. The D. immitis-specific primers confirmed in our previous study and a newly designed TaqMan probe were used for quantitative diagnostics. First, SYBR Green real-time PCR was performed using the specific primers and serially diluted genomic DNA or plasmid DNA, and melting curve analyses were performed after amplification. The melting curve showed one specific peak in each of the genomic and plasmid DNA reactions, suggesting that the primers specifically amplify the D. immitis cytochrome c oxidase subunit I gene. Comparison of SYBR Green and TaqMan real-time PCR using serially diluted plasmid DNA showed higher efficiency and specificity with TaqMan real-time PCR. The real-time PCR assays developed in this study will provide improved diagnostic methods to overcome the limitations of conventional diagnostic tools and facilitate more rapid and accurate diagnoses.

• Biomedical Science Letters
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• v.5 no.1
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• pp.95-100
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• 1999

It is generally difficult, time-consuming, and expensive for the clinical laboratory to detect vancomycin resistant enterococci (VRE). The aim of this study was to develop and evaluate the multiplex polymerase chain reaction (PCR) assay system as a diagnostic tool for the rapid detection of VRE from clinical samples and/or for the identification of VRE from the bacterial strains isolated from clinical specimens. Specific primers, designed from the nucleotide sequences respectively encoding the vanA, vanB, vanC-1, vanC-2/3 genes in enterococci, were coupled in a multiplex PCR assay system. With this multiplex PCR assay system, we investigated the incidence rates and types of VRE isolated from clinical samples. A total of 75 strains of enterococci were isolated in 3 general hospitals in Korea. Of these isolates, 36 strains showed a pattern of high-level vancomycin resistance which associated with vanA gene, whereas 18 strains showed low-level vancomycin resistance associated with vanC-1 or vanC-2/3 gene. Thus, multiplex PCR assay method established in this study could be applied for the rapid detection of VRE.

• 연구논문집
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• s.33
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• pp.17-25
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• 2003

In this work, thermal design of a PCR chip for LOC is systematically conducted. From the numerical simulation of a PCR chip based on the finite volume method, how to control the average temperature of a PCR chip and the temperature difference between the denaturation zone and the annealing zone is presented. The average temperature is shown to be controlled by adjusting heat input and a cooler as well as a heater is shown to be necessary to obtain three individual temperature zones for polymerase chain reaction. To reduce the time required, a heat sink for the cooler is not included in the calculation domain for the PCR chip and heat sink design is conducted separately by using a compact modeling method, the porous medium approach.

• Journal of Genetic Medicine
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• v.5 no.1
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• pp.15-20
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• 2008

Purpose : Large exon deletions in the DMD gene are found in about 60% of DMD/BMD patients. Multiplex PCR has been employed to detect the deletion mutation, which frequently generates noise PCR products due to the presence of multiple primers in a single reaction as well as the stringency of PCR conditions. This often leads to a false-negative or false-positive result. To address this problematic issue, we introduced the dual primer oligonucleotide (DPO) system. DPO contains two separate priming regions joined by a polydeoxyinosine linker that results in high PCR specificity even under suboptimal PCR conditions. Methods : We tested 50 healthy male controls, 50 patients with deletion mutation as deletion-positive patient controls, and 20 patients with no deletions as deletion-negative patient controls using DPO-multiplex PCR. Both the presence and extent of deletion were verified by simplex PCR spanning the promoter region (PM) and 18 exons including exons 3, 4, 6, 8, 12, 13, 17, 19, 43-48, 50-52, and 60 in all 120 controls. Results : DPO-multiplex PCR showed 100% sensitivity and specificity for the detection a deletion. However, it showed 97.1% sensitivity and 100% specificity for determining the extent of deletions. Conclusion : The DPO-multiplex PCR method is a useful molecular test to detect large deletions of DMD for the diagnosis of patients with DMD/BMD because it is easy to perform, fast, and cost-effective and has excellent sensitivity and specificity.

• Proceedings of the KOR-BRONCHOESO Conference
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• 1993.05a
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• pp.87-87
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• 1993

Polymerase chain reaction is widely used as a powerful tool in modern molecular biology. As there is agreement that the HPV is an important factor in the head and neck cancers, the detection of HPV DNA sequence in the head and neck cancer tissue has been tried in several ways. We used the PCR to detect the E1 open reading frames of the HPV in paraffin-embedded tissue of the patients with the head neck cancers. Eleven of the fifty-four tested samples (30%) showed positive result. We have analysed the clinical courses and characteristics related with Human Papillomavirus in those patients.

• Journal of the Korean Chemical Society
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• v.49 no.3
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• pp.255-260
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• 2005

• Journal of the Korean Society for Precision Engineering
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• v.25 no.11
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• pp.37-44
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• 2008