• Korean Journal of Food Science and Technology
• /
• v.36 no.5
• /
• pp.723-727
• /
• 2004

Qualitative and quantitative PCR methods were performed to examine detection and quantitation of epsps inserted into genetically modified soybean (GMS) in processed foods, soy milk, tofu, and biji (soybean curd residue). Using PCR amplification to produce two (121 and 330 bp) epsps in GMS, detection limits of GMS in soy milk, tofu, and biji containing 0.01% GMS were measured. For quantitative detection, test samples containing 1, 3, and 5% GMS were measured by real-time PCR method. Results show real-time PCR method is applicable to detect GMS quantitatively in processed foods.

• Journal of Veterinary Clinics
• /
• v.27 no.5
• /
• pp.508-513
• /
• 2010

Cytokines are important mediators of the immune response, and quantitating cytokine mRNA is a highly sensitive and attractive method for measuring cytokine production. The objective of the current study was to develop and validate a SYBR green quantitative real-time reverse transcriptase PCR (qRT-PCR) assay for measuring canine cytokine mRNA. The optimal annealing temperatures ($T_a$) of the designed primers were $62^{\circ}C$ for interleukin (IL)-$1{\beta}$, IL-6 and IL-10; $60^{\circ}C$ for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and tumor necrosis factor (TNF)-${\alpha}$; and $58^{\circ}C$ for high mobility group box 1 (HMGB1). Primer efficiencies of all primers calculated for standard curve samples were between 97.1% and 102.6%. No evidence of secondary structure or primer-dimer formation was seen via melt-curve analysis or gel electrophoresis. The developed qRT-PCR assays are highly specific and sensitive and can be used to quantify gene expression levels of canine cytokines.

• Proceedings of the Korea Air Pollution Research Association Conference
• /
• 2008.10a
• /
• pp.274-274
• /
• 2008

• Journal of Korean Society of Environmental Engineers
• /
• v.32 no.2
• /
• pp.121-130
• /
• 2010

Metals and sulfate can be considerably dissolved at low pH condition in the acid mine drainage(AMD) and it would make an environmental problems. There are only few of acid mine drainage treatment systems in Korea which are operating, but these still have an effect on the surrounding stream. In this study, quantification of indicator microorganisms was conducted to judge the environmental impact of AMD on microflora by quantitative real-time PCR in the drainage samples of four mines and the water samples of each surrounding stream. Two species of iron reducing bacteria(Rhodoferax ferrireducens T118 and Acidiphilium cryptum JF-5) were selected for indicator bacteria based on 16S rRNA cloning analysis, and sulfate reducing bacteria(Desulfosporosinus orientus), iron and sulfur oxidizing bacteria(Acidothiobacillus ferrooxidans) and iron oxidizing bacteria(Leptosprillum ferrooxidans) were included into indicator since these were found in the previous studies on the mining area. Thereafter, the comparative analysis of four mines were established by the microbiological variation index and it was determined that the biological environment effect of AMD is highest in Samtan mine which doesn t contain treatment system by the value.

• Journal of Food Hygiene and Safety
• /
• v.30 no.1
• /
• pp.103-108
• /
• 2015

Weissella spp. from traditional Korean foods of Kimchi were isolated and characterized against food-borne pathogenic Listeria monocytogens. The isolates were identified as W. cibaria 0D17 and W. confusa 0D23 from Kimchi by the biochemical characteristics and 16S DNA sequencing. The culture solutions of the isolates adjusted to pH 7.0 showed L. monocytogens inhibition. To analyze the quantitative detection of L. monocytogenes, real-time PCR was performed according to the SYBR Green I method. The isolates grew well and L. monocytogens did not grow during the co-culture with those strains at $37^{\circ}C$. Therefore, W. cibaria 0D17 and W. confusa 0D23 might be the candidates as the functional lactic acid bacteria for improving food safety.

• Journal of Apiculture
• /
• v.32 no.2
• /
• pp.99-109
• /
• 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 Food Hygiene and Safety
• /
• v.33 no.1
• /
• pp.71-76
• /
• 2018

Norovirus is a leading cause of sporadic pathogenic non-bacterial gastroenteritis worldwide. For the detection of norovirus, reverse transcription real-time PCR (RT qPCR) has quickly become a major tool due to its sensitivity and specificity. However, accurate viral RNA extraction methods are essential for RT qPCR analysis. TRIzol reagents are used to extract RNA from biological materials and are therefore widely used for norovirus RNA extraction. In this study, the yield of viral RNA extraction using TRIzol from genogroup II (GII) among the human norovirus genogroup I (GI) and GII, and murine norovirus (GV) depended on the pH of the virus sample solution. The yield of RNA extraction was higher at the alkaline pH than in the acidic region compared with the Ct (threshold cycle) value of the real-time PCR. From the results of this study, it was found that the pH condition is very important for the quantitative analysis of norovirus by extracting GII RNA using TRIzol.

• Korean Journal of Microbiology
• /
• v.44 no.1
• /
• pp.49-55
• /
• 2008

White Spot Syndrome Virus (WSSV) is one of the most virulent viral agents in the penaeid shrimp culture industry. In this study, WSSV in a Fenneropenaeus chinensis shrimp farm and an adjacent seawater were concentrated using a membrane filtration and quantified using the quantitative real-time PCR (QRT-PCR) method with newly designed primers and Taqman probe. Sensitivity of primers and probe was proven by WSSV standard curve assay in QRT-PCR. In order to demonstrate the relationship between WSSV and environmental parameters, physicochemical and biological parameters of the farm and influent seawaters were monitored from June to September, 2007. The abundance of WSSV ranged 3,814-121,546 copies per 1 liter of seawater, which was correlated with fecal enterococci ($r^2=0.9$, p=0.02), chlorophyll ${\alpha}$ ($r^2=0.8$, p=0.03) and $BOD_5$ ($r^2=0.8$, p=0.07). Subsequently, it is concluded that the QRT-PCR method using Taqman probe established in this study was efficient to clarify the quantification of WSSV in seawaters. Statistical analyses of environmental parameters obtained in this study also showed that the abundance of WSSV was correlated with several biological parameters rather than physicochemical parameters.

• Journal of Veterinary Clinics
• /
• v.22 no.4
• /
• pp.348-352
• /
• 2005

We described a rapid, sensitive and reproducible real-time PCR assay for detection and quantitation of canine parvovirus type 2 in the feces of dogs with parvovirus infection. The method was demonstrated to be highly specific and sensitive, allowing a precise canine parvovirus type-2 quantitation over range of eight orders of magnitude from $10^2\;to\;10^9$ copies of standard DNA. Then, fecal samples from parvovirus infected dogs were analyzed by conventional PCR and real-time PCR. Real-time PCR is more sensitive than conventional PCR, allowing to detect low viral titers of CPV-2 in infected dogs. By real-time PCR, a wide range of parvovirus particles was found in the samples from $1.45\times10^6\;to\;9.45\times10^8$ copies/0.01g of feces. However, when dogs are in infection of parvovirus, it is difficult to prove that the numbers of peripheral blood leukocytes are correlated with those of fecal shedding virus.

• 보존과학연구
• /
• s.32
• /
• pp.171-183
• /
• 2011

In this study molecular genetic analysis was carried out on 4 human skeletal remains from Osuri, Buyeo. We showed that real-time PCR is the method of the choice to assess the initial number of genuine ancient DNA molecules. Human mitochondrial DNA quantification was accomplished by the real-time PCR for the cytochrome b gene of the mitochondria. Histological results proved to be a good potentiality for biochemical analysis using biomolecule. The level of specimen's preservation state was proved that level of quantitative result was BO-04, BO-01, BO-03, BO-02. Continually, we showed that biochemical and biomolecule results for the level of preservation state were similar. This study will be useful to important material for predicting biochemistry and biology analysis of the ancient bone.