• 대한임상검사과학회지
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• 제39권3호
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• pp.147-150
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• 2007

Most expressed HLA (human leukocyte antigen) loci exhibit a remarkable degree of allelic polymorphism, which derives from sequence differences predominantly localized to discrete hypervariable regions of the amino terminal domain of the molecule. In this study, the HLA-B genotypes were determined in twenty students unrelated koreans using the PCR-SSP (polymerase chain reaction-sequence specific primer) technique. Several specific primer pairs in assigning the HLA-B gene were used ($B^{\ast}4001/4007$, $B^{\ast}4901/5001/4501$, $B^{\ast}3701$, $B^{\ast}5801$). The results of PCR-SSP, the HLA-B3701 primer was detected one (5%), the $HLA-B^{\ast}5801$ were detected four (20%), the $HLA-B^{\ast}4001/4007$ were detected nineteen (95%) and the $HLA-B^{\ast}4901/5001/4501$ were detected twenty. This study shows that the PCR-SSP technique is relatively simple, fast and a practical tool for the determination of the HLA-B genotypes. Moreover, these results genotype frequency of the HLA-B gene could be useful for database study before being applied to individual identification and transplantation immunity.

• Journal of Microbiology
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• 제39권3호
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• pp.226-228
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• 2001

Human lymphotropic herpesvirus is known to be a major pathogen associated with various diseases in bone marrow transplantation (BMT) recipients. A multiplex nested-polymerase chain reaction (PCR) method was developed for the simultaneous detection of human lymphotropic herpesviruses, including Ebstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 variants A and B (HHV6-A, HHV6-B). To demonstrate the usefulness of multiplex PCR for the analysis of clinical samples, peripheral blood mononuclear cells and serum from BMT recipients were analysed. The results skewed that a clear detection could be made between EBV, HCMV and HHV-6. This multiplex PCR assay is an efficient and cost-effective approach to the analysis of large numbers of samples to determine the epidemiological importance of EBV HCMV and HHV-6.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.4989-4992
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• 2013

Aim: To compare p16INK4a immunocytochemistry with the HPV polymerase chain reaction in predicting high grade cervical squamous intraepithelial lesions. Materials and Methods: This diagnostic case-control study was conducted from January 2010 until December 2010. We obtained 30 samples, classified according to the degree of cervical intraepithelial neoplasia (CIN): 11 samples for CIN 1, 9 samples for CIN 2, and 10 samples for CIN 3. HPV PCR, p16INK4a immunocytochemistry, and histopathological examination were performed on all samples. Statistical analysis was conducted using SPSS 20.0. Results: In predicting CIN 2-3, we found p16INK4a to have similar specificity and positive predictive value as HPV PCR (95%, 97.2% vs 96.7%), but better sensitivity (87.5% vs 72.5%) and negative predictive value (82.1% vs 67.6%). The most prevalent types of high-risk HPV in our study were HPV 33, 35, 58, 52, and 16. Conclusions: p16INK4a has better diagnostic values than HPV PCR and may be incorporated in the triage of ASCUS and LSIL to replace HPV PCR. Genotype distribution of HPV differs in each region, providing a challenge to develop HPV vaccines based on the epidemiology of HPV in that particular region.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.801-806
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• 2002

A multiplex competitive polymerase chain reaction (PCR) method was developed for the rapid identification and quantification of Leuconostoc mesnteroides and Lactobacillus plantarum populations which are the key microorganisms in kimchi fermentation. The strain-specific primers were designed to selectively amplify the target genes encoding 165 rRNA of L. plantarum and dextransucrase of L. mesenteroides. There was a linear relationship between the band intensity of PCR products and the number of colony forming units of each model organism. The PCR quantification method was compared with a traditional plate-counting method f3r the enumeration of the two lactic acid bacteria in a mixed suspension culture and also applied to a real food system, namely, watery kimchi. The population dynamics of the two model organisms in the mixed culture were reliably predictable by the competitive PCR analysis.

• 한국동물위생학회지
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• 제24권1호
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• pp.43-50
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• 2001

Swine proliferative enteritis(SPE) caused by inかsoma intracellularis is a common enteric disaese of grower and finisher pig. Swine affected with SPE show variable clinical signs including diarrhea, weight loss, aberrant growth and death. The characteristic lesion of ileitis at necropsy is marked thickening of the last section of the small intestine. The inner lining of the thickened intestine proliferates almost like a cancer and curved rod bacteria(L intracellularis) are always seen inside the intestinal wall. Infected swine shed the organism in the feces. Isolation and growth of pure L intracellularis in vitro requires a suitable cell culture. This procedure is difficult and not a practical means of diagnosis, thus the polymerase chain reaction(PCR) test of feces can be used to determine whether a pig is shedding the infective organism. A sensitive assay based on amplification of a 319bp DffA fragment of the L intracellularis of Swine proliferative enteritis was attempted for the detection of the organism in the 62 feces of swine. L intracellularis was identified on three herds and detected in 6 fecal samples, representing a infection rate of 9.7%. The PCR was very sensitive and specific on the individual level. The PCR technique could be very useful for the diagnosis of this disease.

• 센서학회지
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• 제14권5호
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• pp.337-342
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• 2005

A silicon-based micro-reactor to amplify small amount of deoxyribonucleic acid (DNA) has been fabricated using micro-electro-mechanical systems (MEMS) technology. Polymerase chain reaction (PCR) of DNA requires a precise and rapid temperature control. A Pt sensor is integrated directly in the chamber for real-time temperature measurement and an infrared lamp is used as external heating source for non-contact and rapid heating. In addition to the real-time temperature sensing, PCR needs a rapid thermocycling for effective PCR. For a fast thermal response, the thermal mass of the reactor chamber is minimized by removal of bulk silicon volume around the reactor using double-side KOH etching. The transparent optical property of silicon in the infrared wavelength range provides an efficient absorption of thermal energy into the reacting sample without being absorbed by silicon reactor chamber. It is confirmed that the fabricated micro-reactor could be heated up in less than 30 sec to the denaturation temperature by the external infrared lamp and cooled down in 30 sec to the annealing temperature by passive cooling.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1171-1177
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• 2008

This study was undertaken to classify Enterobacter sakazakii isolates from 13 powdered infant formula products, 25 powdered weaning diet products, and 33 weaning diet ingredients on polymerse chain reaction (PCR) methods. The numbers of the isolates from 1 powdered infant formula product, 7 powdered weaning diet products, and 6 weaning diet ingredients were 1, 14, and 8, respectively. The contaminated ingredients were 1 rice powder, 2 millet powders, 2 vegetable powders, and 1 fruit and vegetable premix. PCR with the primer of repetitive extragenic palindromic element (REP-PCR) and random amplification of polymorphic DNA(RAPD) were effective in discriminating among the isolates, but tRNA-PCR and PCR with the primer of l6S-23S internal transcribed spacer (ITS-PCR) were not. Some of E. sakazakii isolates from vegetable powders, fruit and vegetable premix, and millets powders were classified into the clonal groups based on the DNA patterns in the REP-PCR and RAPD analysis. A close genetic relationship among the isolates from some of the powdered weaning diet products and the rice powder was also detected in the cluster analysis based on the DNA patterns in RAPD.

• 한국식물병리학회지
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• 제14권4호
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• pp.308-313
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• 1998

Reverse transcription and polymerase chain reaction (RT-PCR) techiniques were used to identification and differentiation of cucumber mosaic virus isolated from Forsythia koreana (CMV-Fk). RT-PCT used by two set of 20-mer primers one was CMV-common primers and another was CMV subgroup I-specific primers designed in a conserved region of the 3' end of CMV RNA3, amplified about 490 bp and 200 bp DNA fragments from CMV-Fk, respectively. CMV could be detected by RT-PCR at a dilution as low as 10-4 in forsythia crude sap extracts. Restriction enzyme analysis of RT-PCR products using EcoRI and MspI showed that CMV-Fk belonged to CMV subgroup I. But, analysis of RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR) showed heterogeneity of RNA3 between CMV-Fk and CMV-Y as a member of subgroup I.

• Reproductive and Developmental Biology
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• 제31권1호
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• pp.55-60
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• 2007

Aging causes thymus involution, and genes in thymus play an important role in the development of the immune system. In this study, we compared genes expressed in thymus of neonatal and peripubertal rats using annealing control primers (ACPs)-based GeneFishing polymerase chain reaction (PCR) and semiquantitative reverse transcription (RT)-PCR. We identified 10 differentially expressed genes (DEGs) with 20 ACPs. Of 10 DEGs, bystin-like, collagen type V alpha 1 (COL5A1), and T-cell receptor beta-chain segment 2 (TCRB2) that are related to immune-function were detected in rat thymus. Bystin-like and TCRB2 were up-regulated, while COL5A1 was down-regulated in peripubertal thymus. Semiquantitative RT-PCR confirmed postnatal changes in expression of bystin-like, COL5A1, and TCRB2. These results suggest that bystin-like, COL5A1, and TCRB2 could regulate immune function controlled in thymus as age increases.

• 한국양식학회지
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• 제10권2호
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• pp.171-178
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• 1997

Infectious pancreatic necrosis virus (IPNV)는 치어에 감염되어 치명적인 질병을 유발하는, 양식산업에 있어 중요한 어류 병원체이다. 본 연구에서는 IPNV를 신속, 정확하게 진단하는 방법을 개발하고자 IPNV의 항원성 단백질인 VP2 유전자 부분에서 선택한 primers를 이용하여 역전사-중합효소연쇄반응법(Reverse Transcription-Polymerase Chain Reaction, RT-PCR)을 실시하였다. RT-PCR 증폭법으로 순수분리도니 IPNV dsRNA 40 ng 정도의 적은 양도 확인 할 수 있었으며, IHNV와 같은 다른 어류 병원체의 게놈을 RT-PCR templates로 사용하였을 경우는 어떠한 PCR 산물도 검출되지 않는 특이성을 보였다. 특히 유전자의 분리없이 조직 그 자체를 대상으로 RT-PCR을 행하는 in situ RT-PCR 방법으로 IPNV가 감염된 넙치 (Paralichthy olivaceus) 치어의 조직에서 IPNV 감염을 신속하게 확인할 수 있었다. 따라서 RT-PCR 및 in situ RT-PCR 방법은 IPNV를 신속, 정확하게 진단하는데 활용될 수 있을 것으로 생각된다.