• Journal of Plant Biotechnology
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• v.38 no.2
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• pp.143-150
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• 2011

Since the first commercial GM plant, the FlavrSavr tomato, authorized in 1994, more than 140 GM plants were authorized for marketing globally. For the authorization and labelling of GM plants, the detection methods for genes introduced and proteins expressed in GM plants were developed qualitatively and quantitatively. This review presented the detection methods, conventional PCR, multiplex PCR and real-time PCR, for soybean, maize, canola and cotton as the dominant GM plants. Also, microarray assay and nanotechnology as new approaches for detection methods for GM plants were investigated.

• Journal of Apiculture
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• v.32 no.2
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• pp.99-109
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• 2017

The multiple real-time PCRs against pathogens of Bombus species including DWV, IAPV, KBV, SBV, BQCV, kSBV, SBPV and Paenibacillus larvae, Mellisococcus plutonius, Lysinibacillus fusiformis, and Klebsiella oxytoca have been developed. One extracted nucleic acid from Beesample could be applied to 11 different PCRs in same time and condition. Specific PCR-products were amplified qualitative and quantitative manner inner 20 minutes successfully, when each 1000 molecules of pathogen-specific target DNA is existed as template, respectively. The multiple PCR detection that we propose would be expected to apply to quarantine test for international exchange of Bombus species.

• Journal of fish pathology
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• v.16 no.3
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• pp.161-164
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• 2003

An Infectious hematopoietic necrosis virus strain (IHNV-RTK) was isolated from cultured rainbow trout at Kumi and Jechun area in Korea during 2000 and 2001. In the RT-PCR amplification with the specific primer set designed from IHNV G protein region, a 540 bp PCR product was amplified from the RTK strain. The RTK strain showed higher sequence homology with the published IHNV G protein genes (RB-76, LR-73, Col-85, and Carson-89)

• Journal of the Korean Chemical Society
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• v.51 no.1
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• pp.36-42
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• 2007

Hepatitis B is a serious public health problem leading to chronic infection and liver cancer. Quantitation of circulating hepatitis B virus (HBV) is important for monitoring disease progression and for assessing the response to antiviral therapy. In this study, by using Real-Time PCR and novel Micro-PCR assay method, we measured HBV concentration in the clinical sample. A total of 120 serum samples from patients with HBV infection collected was in Dankook university hospital to compare the detection limit, sensitivity, specificity and reproducibility of the two assay methods. These findings of this study suggest that Micro-PCR and Real-Time PCR assay methods are comparable to each other in there detection limit, sensitivity, and reproducibility for HBV DNA quantitation. However, Micro-PCR assay is more efficient than Real-Time PCR method, because Real-Time PCR is not so time - consuming, technically easy and need to reagent of a small quantity. It will be useful for rapid and reliable clinical diagnosis of HBV in many countries.

• Journal of Korean society of Dental Hygiene
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• v.17 no.1
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• pp.13-25
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• 2017

Objectives: The purpose of this study was to compare colony forming unit (CFU) method and multiplex real-time polymerase chain reaction (MRT-PCR) method for accurate quantitative analysis of bacteria. Methods: We compared the CFU method and the MRT-PCR method, which are still used in Korea, for Prevotella intermedius (P. intermedius), a periodontal disease pathogen selected by MRT-PCR, and Streptococcus mutans (S. mutans), a dental caries causative organism. The subjects of this study were 30 patients who visited the C dental hospital. Results: Total microorganisms in MRT-PCR method were significantly higher in both types of bacteria (p<0.05), since DNA of dead bacteria was also analyzed. This was because the periodontal dise(-) anaerobes, and even dead bacteria contain large amounts of toxic substances called LPS in the extracellular membrane, and fimbriae and pili, which are motility structures, still remain as a strong toxic substance in periodontal tissue. Conclusions: Therefore, in terms of the total amount of bacteria found, the MRT-PCR method will be a useful technique for searching all the bacteria in the oral cavity including live bacteria, as well as sterilization.

• Journal of fish pathology
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• v.24 no.2
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• pp.65-73
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• 2011

In analysis of DNA viruses from the contaminated shellfish using PCR, preparation method of template DNA is an important factor to get enough copy number of viruses. In this study, we evaluated the efficiency of PCR template of Megalocytivirus (sT50mg-D) DNA obtained from 50 mg digestive gland homogenate of oyster using commercial method, and compared with that obtained from 5 g of the same tissues (T5g-D) after PEG precipitation procedures of virus. Both templates DNA suspended in the same volume of distilled water showed positive results by primary PCR with 35 cycles, and the presence of Megalocytivirus was confirmed in oysters collected from cultured farms in Korea. Moreover, PCR with sT50mg-D allowed us to discriminate the contaminated oyster individually, that can not be done in PCR with T5g-D prepared from the mixture of three different individual oyster to get 5 g digestive gland homogenate. In quantitative analysis with real time PCR, Megalocytivirus concentrations in 50 ${\mu}l$ templates prepared using 0.5~50 mg of one positive sample were appeared in the range 6.14E+00~1.2E+02/${\mu}l$. We were not able to get positive result using template DNA contained less than 6.14E+00 copies. Consequently, 2-step PCR performed with DNA extracts from oyster homogenate of small amount (sT50mg-D) i) was enough to detect the contaminated Megalocytivirus in shellfish, ii) allowed us to do the analysis for individual shellfish rather than mixture of several shellfish and iii) showed the presence of Megalocytivirus in oyster from Korea.

• Journal of Aquaculture
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• v.17 no.1
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• pp.24-29
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• 2004

The high mortality of cultured turbot, Scophthalmus maximus was occurred from Gochang in June, 2003. Infect fish with iridovirus showed a lower feed intake and lethargic. These fish exhibited pale body color, extended abdominal and exophthalmus. Histopathological studies showed basophilic enlarged cells from kidney, spleen, gill, heart, muscle, stomach, intestine, liver and pancreas of these fish. In PCR amplification with red sea bream iridovirus (RSIV) specific primer set of a DNA fragment of 286 bp was obtained from infect turbot with the virus. The strain showed the high homologies with RSIV, LBIV (largemouth bass iridovirus), GSIV (giant seaperch iridovirus) and SBIV (sea bass iridovirus).

• KSBB Journal
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• v.21 no.1 s.96
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• pp.59-67
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• 2006

2D spectrofluorometer produces many spectral data during fermentation processes. The fluorescence spectra are analyzed using chemometric methods such as principal component analysis (PCA), principal component regression (PCR) and partial least square regression (PLS). Analysis of the spectral data by PCA results in scores and loadings that are visualized in score-loading plots and used to monitor a few fermentation processes by S. cerevisae and recombinant E. coli. Two chemometric models were established to analyze the correlation between fluorescence spectra and process variables using PCR and PLS, and PLS was found to show slightly better calibration and prediction performance than PCR.

• Journal of Life Science
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• v.20 no.12
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• pp.1896-1901
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• 2010

This study was conducted using quantitative real-time PCR using Lactobacilli as probiotics. Quantitative real-time PCR (RT PCR) was conducted via a method involving SYBR Green 1 and a probe. Plasmid DNA was cloned using the 16S-23S rRNA intergenic species region. Gene clones were diluted from $10^2$ to $10^{10}$. Standard curves were constructed via Ct values obtained from the results of Real-time PCR via the aforementioned SYBR Green 1 and probe method. Plasmid DNA was also cloned using the 16S-23S rRNA intergenic species region and the gene clones were diluted from $10^2$ to $10^{10}$ copy numbers via the probe method. Using RT PCR, a standard curve of plasmid DNA copy numbers was also determined. The slope value for the Y-axis intercept and $R^2$ value were measured as -3.346, 33.18, and 0.993, respectively, via the first method. For the second method, the slope value for the Y-axis intercept and $R^2$ were -3.321, 31.10 and 0.995, respectively. The PCR inhibitor could not express the detection curve at a copy number over $10^{10}$ via either method, owing to high DNA density. The DNA extract from probiotics was diluted without pre-culturing, and 16 products were amplified via both methods. The Ct value was 11.06~18.12 in the first method and 16.74~22.11 in the second method. Measured probiotics and log copy values were largely similar among the methods used. It was concluded that both methods are effective for analysis, but further research will be required to verify the optimal method.

• Korean Journal of Food Science and Technology
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• v.35 no.4
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• pp.591-597
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• 2003

To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also, to remove an excessive carbohydrate from the potato, ${\alpha}$- and ${\beta}$-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted from the raw potato with high purity for PCR. However, the silica-coated head method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNA was extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCR efficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2% agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.