• Parasites, Hosts and Diseases
• /
• v.37 no.4
• /
• pp.289-292
• /
• 1999

BALB/c mice infected with a high virulent strain of Trypanosoma brucei gambiense IL3707 were treated intraperitoneally (ip) with either Melarsoprol （Mel-B) or PSG(+) buffer as controls. The mice were subsequently monitored regularly for parasites by direct microscopic examination of their tail blood or buffy coat and by polymerase chain reaction (PCR). Mel-B was found to be an effective drug for treatment against T.b. gambiense because at the end of the first treatment schedule, all treated mice were negative for parasites even by PCR, while all the control animals were positive. Three of the five Mel-B treated mice, while parasitologically negative, were PCR positive between 53 and 80 days post infection (DPI), indicating that they still harbored an infection. All treated mice were subsequently negative for parasites even by PCR at 88 DPI. A combination of conventional microscopic examination and PCR offers a good prediction of cure following treatment of trypanosomosis.

• Analytical Science and Technology
• /
• v.12 no.3
• /
• pp.260-264
• /
• 1999

A simple and rapid procedure, called FoLT-PCR(Formamide Low Temperature-Polymerase Chain Reaction) was applied to amplifying DNA directly from various forensic biological evidences including human blood, saliva, hair root, or semen without any DNA preparative steps. We added washing step with non-ionic detergent, 1% Triton X-100, and used Taq DNA polymerase instead of Tth DNA polymerase to amplify 3 STR loci and gender allele simultaneouly. Optimal concentration of formamide and annealing temperature were determined empirically to 8%(v/v), and $48^{\circ}C$ respectively. We also compared this method with standard PCR.

• Korean Journal of Pharmacognosy
• /
• v.30 no.4
• /
• pp.355-362
• /
• 1999

New lectins have been isolated and purified from mung bean (Phaseolus radiatus) through physiological saline extraction, ammonium sulfate salt fractionation and column chromatographies. Ion exchanger were eluted by linear salt gradient and then further purified through gel filtration. Thus obtained lectin named as MBL. The gene expressions of 5 cytokines (IL-1, IL-2, IL-6, $TNF-{\aphpa}$ and $IFN-{\gamma}$) from human peripheral blood mononuclear cells (PBMC) stimulated with MBL were investigated by using reverse transcription polymerase chain reaction (RT-PCR). PBMC ($1{\times}106$ cells/ml) isolated from healthy volunteers were stimulated with lectins (4 mg/ml) for various time intervals (1 to 96 hrs). After each of the various stimulated times, total RNA was isolated and assessed for different cytokines mRNA by RT-PCR. The mRNA encoding IL-1, IL-2 were detected continuously from 1 to 20 hrs, and IL-6 was detected up to 24 hrs. But the mRNA encoding $IFN-{\gamma}$ and $TNF-{\alpha}$ were detected to 8 hours only and showed short time response compared with other cytokines. The significant expression of all cytokines mRNA were observed at 4 hrs. These results suggested that MBL, as inducer of cytokines could elicit detectable cytokine mRNA from PBMC.

• Korean Journal of Ecology and Environment
• /
• v.54 no.3
• /
• pp.199-208
• /
• 2021

Environmental DNA (eDNA) metabarcoding is effective method with high detection sensitivity for evaluating fish biodiversity and detecting endangered fish from natural water samples. We compared the richness of operational taxonomic units(OTUs) and composition of freshwater fishes according to filters(cellulose nitrate filter vs. glass fiber filter), extraction kits(DNeasy2^® Blood & Tissue Kit vs. DNeasy2^® PowerWater Kit), primer sets (12S rDNA vs. 16S rDNA), and PCR methods (conventional PCR vs. touchdown PCR) to determine the optimal conditions for metabarcoding analysis of Korean freshwater fish. The glass fiber filter and DNeasy2^® Blood & Tissue Kit combination showed the highest number of freshwater fish OTUs in both 12S and 16S rDNA. Among the four types, the primer sets only showed statistically significant difference in the average number of OTUs in class Actinopterygii (non-parametric Wilcoxon signed ranks test, p=0.005). However, there was no difference in the average number of OTUs in freshwater fish. The species composition also showed significant difference according to primer sets (PERMANOVA, Pseudo-F=6.9489, p=0.006), but no differences were observed in the other three types. The non-metric multidimensional scaling (NMDS) results revealed that species composition clustered together according to primer sets based on similarity of 65%; 16S rDNA primer set was mainly attributed to endangered species such as Microphysogobio koreensis and Pseudogobio brevicorpus. In contrast, the 12S rDNA primer set was mainly attributed to common species such as Zacco platypus and Coreoperca herzi. This study provides essential information on species diversity analysis using metabarcoding for environmental water samples obtained from rivers in Korea.

• Biomedical Science Letters
• /
• v.24 no.2
• /
• pp.76-86
• /
• 2018

Breast cancer is one of the most common cancers affecting women worldwide. Although the survival rate of breast cancer has increased, breast cancer still results in a high mortality rate. Breast cancer deaths are caused by metastasis that occurs in organ dysfunction. Recently, there have been many studies on circulating tumor cells (CTCs), which are related to breast cancer metastasis in the blood. Recent studies have demonstrated that some CTCs do not express epithelial markers. Therefore, in this study, total RNA was extracted from blood without separating out the CTCs, and the characteristics of the CTCs were analyzed by RT-qPCR. Cyclin D1 and twist-related protein 1 (TWIST1) are well-known markers for predicting the prognosis of patients with breast cancer. However, few studies have demonstrated the use of CCND1 and TWIST1 in blood as diagnostic and prognostic markers of breast cancer. In this study, patients with late-stage breast cancer had overexpressed CCND1 and TWIST1 than patients with different stages of breast cancer (P < 0.001 and P < 0.01, respectively). The relative expression level of CCND1 in survivors was higher than in patients who died (P = 0.06). The relative expression level of TWIST1 in survivors was lower than in patients who died (P = 0.08). Overall CCND1 and TWIST1 were not useful as markers for the diagnosis of breast cancer through blood. However, we showed the possibility of using CCND1 and TWIST1 as prognostic markers, and a large-scale study is needed to confirm the usefulness of these prognostic markers.

• Journal of Biosystems Engineering
• /
• v.38 no.2
• /
• pp.121-128
• /
• 2013

Purpose: Porcine proliferative enteropathy (PPE), caused by the obligate intracellular bacterium Lawsonia intracellularis, is a widely distributed disease throughout the world causing substantial economic loss. In order to diagnose PPE rapidly, the rapid kit was developed and tested. Methods: In this study, a rapid kit was developed to screen the PPE rapidly at the pig farm. Also, occult blood test with fecal occult blood (FOB) kit was done for detecting the blood in pig feces which might be the evident of hemorrhagic PPE. For developing the kit, we tested fecal samples of PPE infected pigs diagnosed by polymerase chain reaction (PCR) method. Results: With the developed rapid kit, Lawsonia intracellularis was detected in high density emulsion of ileum. On the other hand, the test result of detecting Lawsonia in feces showed too high non-specific response. In addition, nevertheless the FOB test result showed that blood evident could be founded in pig feces, the diagnosing result was not fit to PCR test result, which shows blood in pig feces could be from not only hemorrhagic PPE but also many reasons. Conclusions: To deal with the PPE effectively, it will be better for farmers to screen the PPE in earlier stage with easy and rapid diagnosing tool on farm. This study found out that the rapid kit could detect the Lawsonia intracellularis and hemoglobin in pig feces. However, the non-specific response to negative samples of PPE was too high to use at a pig farm. Further research is needed for lowering the non-specific response with the rapid kit.

• Journal of Life Science
• /
• v.16 no.2 s.75
• /
• pp.274-281
• /
• 2006

A fibrinolytic enzyme gene was isolated from Bacillus subtilis BB-1 by PCR method. Primers for PCR cloning were designed according to pre-identified gene for fibrinolytic enzymes from B. subtilis. The primer sequences were 5'-CGG ATC CGT GAG AGG CAA AAA GGT G-3' and 5'-TGA ATT CTT AAT GTG CTG CTG CTT GTC C-3' as concensus sequences of the fibrinolytic genes of Bacillus species. The PCR product was 1,145 bp and the sequence homology was 99% with nattokinase gene isolated from Japanese natto. The cloned fibrinolytic gene was reconstructed in Bacillus-E. coli shuttle vector, pEB for bulk-production. The fibrinolytic enzyme was purified by FPLC from the cloned B. subtilis 168. The optimum pH and temperature of the enzyme were 7.0 and $35^{\circ}C$, respectively. The fibrinolytic enzyme did not show any activity toward to skim milk, gelatin, casein and blood agar plate. The enzyme specific polyclonal antibody was prepared in rabbit for further assays such as detection of the gene expression in plant cells. This means that the enzyme may be used for health-care such as thrombosis without any hamful effects in the blood vessel.

• Korean Journal of Environmental Biology
• /
• v.35 no.4
• /
• pp.468-475
• /
• 2017

This study is for the consideration of the existence tendency of Kudoa septempunctata in olive flounder. In general, muscle has shown a strong PCR positive reaction in spores containing tissues rather than non-containing tissues. However, blood PCR results showed opposed tendency. In various organs of the tested fish containing spores in muscle tissue, heart had shown positive reaction along with muscle at PCR analysis. Muscle fiber necrosis was observed at the histological observation, and this degeneration was common in both samples. The one sample was the PCR positive muscle containing spore and the other was the PCR positive muscle non-containing spore. Both of muscle tissues indicated a positive reaction at ISH (in-situ hybridization) against K. septempunctata.

• Parasites, Hosts and Diseases
• /
• v.52 no.4
• /
• pp.443-447
• /
• 2014

Babesiosis is an emerging tick-borne disease in humans worldwide; however, little is known about the frequency of infection or prevalence of this disease in other parts of the world, excluding North America. In this study, we aimed to investigate Babesia microti infection frequency in a human population in Mongolia. One hundred blood samples were collected from stock farmers living in Khutul city of Selenge province, Mongolia. The sera and DNA from blood samples were evaluated for the presence of B. microti infection by using indirect fluorescent antibody (IFA) tests and PCR. The positive detection rates obtained using the IFA tests and PCR assays were 7% and 3%, respectively. This study is the first to detect of B. microti infections based on antibody seroprevalence or PCR assays for the presence of B. microti DNA in a Mongolian population.

• Korean Journal of Agricultural Science
• /
• v.38 no.1
• /
• pp.45-50
• /
• 2011

This study was conducted to investigate the farm situation about bovine leukemia virus(BLV) infection that greatly influence productivity in dairy cattle and compare the accuracy of diagnosis for BLV infection between PCR and ELISA techniques. Blood samples of 193 heads from 5 herds in Chungnam and Chungbuk area were used to analyze BLV gene and serum, and the results were obtained as follows. The amplified BLV gene in dairy cattle by PCR technique resulted in 226 bp, 596 bp and 434 bp, respectively, for gag, pol and env, which were well amplified. The infection rates of BLV virus diagnosed by PCR and ELISA techniques ranged from 80.55 to 100% and from 22.22 to 86.95%, respectively, and the infection rates among 5 herds were significantly different in both methods (P<0.05). Further, the average infection rates of 5 herds were 87.05 and 63.21%, respectively, for PCR and ELISA techniques. Kappa statistics for examining consistency of diagnosis by PCR and ELISA techniques showed 0.246, which represents low consistency. Consequently, PCR based BLV technique was considered as a corrective measure for diagnosis of BLV infection in Holstein dairy cattle.