• Journal of Apiculture
• /
• v.34 no.1
• /
• pp.39-46
• /
• 2019

BQCV multi-point PCR was developed as a rapid multiplex detection method for BQCV, one of the viral pathogens of honeybees. It could detect BQCV specific genes qualitative as well as quantitative detection based on ultra-rapid PCR. Three primer pairs (RNA dependent RNA polymerase, capsid protein, 3C like protease) were specifically designed for accurate the detection and were optimized for minimizing the detection time and increasing the sensitivity. Our advanced diagnostic system have the accuracy by lowering the concern about the variation in the BQCV detection site. In addition, it should be an opportunity to identify mutations that are mixed with other viruses.

• Journal of Veterinary Science
• /
• v.24 no.3
• /
• pp.40.1-40.6
• /
• 2023

Analysis of the VP1 gene sequence of the foot and mouth disease virus (FMDV) is critical to understanding viral evolution and disease epidemiology. A standard set of primers have been used for the detection and sequence analysis of the VP1 gene of FMDV directly from suspected clinical samples with limited success. The study validated VP1-specific degenerate primer-based reverse transcription polymerase chain reaction (RT-PCR) for the qualitative detection and sequencing of serotype O FMDV lineages circulating in India. The novel degenerate primer-based RT-PCR amplifying the VP1 gene can circumvent the genetic heterogeneity observed in viruses after cell culture adaptation and facilitate precise viral gene sequence analysis from clinical samples.

• Journal of Veterinary Science
• /
• v.25 no.2
• /
• pp.28.1-28.11
• /
• 2024

Background: Porcine circovirus type 2 (PCV2) infection is ubiquitous around the world. Diagnosis of the porcine circovirus-associated disease requires clinic-pathological elements together with the quantification of viral loads. Furthermore, given pig farms in regions lacking access to sufficient laboratory equipment, developing diagnostic devices with high accuracy, accessibility, and affordability is a necessity. Objectives: This study aims to investigate two newly developed diagnostic tools that may satisfy these criteria. Methods: We collected 250 specimens, including 170 PCV2-positive and 80 PCV2-negative samples. The standard diagnosis and cycle threshold (Ct) values were determined by quantitative polymerase chain reaction (qPCR). Then, two point-of-care (POC) diagnostic platforms, convective polymerase chain reaction (cPCR, qualitative assay: positive or negative results are shown) and EZtargex (quantitative assay: Ct values are shown), were examined and analyzed. Results: The sensitivity and specificity of cPCR were 88.23% and 100%, respectively; the sensitivity and specificity of EZtargex were 87.65% and 100%, respectively. These assays also showed excellent concordance compared with the qPCR assay (κ = 0.828 for cPCR and κ = 0.820 for EZtargex). The statistical analysis showed a great diagnostic power of the EZtargex assay to discriminate between samples with different levels of positivity. Conclusions: The two point-of-care diagnostic platforms are accurate, rapid, convenient and require little training for PCV2 diagnosis. These POC platforms can discriminate viral loads to predict the clinical status of the animals. The current study provided evidence that these diagnostics were applicable with high sensitivity and specificity in the diagnosis of PCV2 infection in the field.

• Korean Journal of Food Science and Technology
• /
• v.52 no.1
• /
• pp.40-45
• /
• 2020

Over 500 genetically modified organisms (GMOs) have been developed since 1996, of which nearly 44% have glufosinate herbicide-tolerant traits. Identification of specific markers that can be used to identify herbicide-tolerant traits is challenging as the DNA sequences of the gene(s) of a trait are highly variable depending on the origin of the gene(s), plant species, and developers. To develop specific PCR marker(s) for the detection of the glufosinate-tolerance trait, DNA sequences of several pat or bar genes were compared and a diverse combination of PCR primer sets were examined using certified reference materials or transgenic plants. Based on both the qualitative and quantitative PCR tests, a primer set specific for pat and non-specific for bar was developed. Additionally, a set of markers that can detect both pat and bar was developed, and the quantitative PCR data indicated that the primer pairs were sensitive enough to detect 0.1% of the mixed seed content rate.

• Toxicological Research
• /
• v.20 no.2
• /
• pp.101-108
• /
• 2004

The different social reflections about the benefits and the potential risks of genetically modified (GM) crops have evolved with .different reactions in different countries. Many countries including Korea are working toward setting down new guidelines. Korea requires companies to label all food that contains more than 3% GM ingredients. One of the rapid and convenient detection methods of GM ingredients is amplification of the introduced DNAs by polymerase chain reaction (PCR). Many PCR kits for this purpose are commercially available. The objective of this study was to evaluate performance of commercialized GM crop detection kits. The results showed that 6 out of 15 kits tested did not meet the requirements even purposed by the manufacturers themselves in terms of stability, reproducibility, and detection limits, suggesting a potential quality control problem in their design stage or production line. The evaluation also suggests that, although the duplex and triplex detection kits allowed unambiguous detection in a single PCR reaction, the monoplex detection kits were the most sensitive to the detection of GM ingredients. The detection limits also differ between soybean and corn. Results from this study will be useful in the development of sound qualitative tracking systems of GM ingredients for monitoring throughout the cultivation of GM crops, their trans-boundary movement, and food production using GM grains as well as for complying with government guidelines associated with GM crops.

• Korean Journal of Environmental Agriculture
• /
• v.26 no.4
• /
• pp.319-324
• /
• 2007

This study was conducted to evaluate the persistence of DNA in the transgenic chili pepper resistant to cucumber mosaic virus (CMV) in the field condition. We analyzed the persistence of genes in the leaf samples obtained from two field conditions, below and above soil. Transgenic and non-transgenic leaf tissues were buried in the soil at a depth of 10 cm or placed on the soil surface. Qualitative and quantitative PCR analysis showed that the amount of transferred genes from the transgenic peppers below and above soil was dropped to 28.3-42.7% one month after buried and it was rapidly reduced to 0.9-3.3% after two months. The transgenes were not detected three to four month after buried. In addition, DNA of the leaves placed below soil decomposed about 8%more than those on soil surface. The gene transfer from decomposing leaves of the transgenic pepper to soil was investigated by PCR analysis with the soil attached to the samples. The PCR result indicated that the gene transfer from the transgenic pepper to soil was not occurred.

• Journal of Dental Rehabilitation and Applied Science
• /
• v.34 no.3
• /
• pp.186-195
• /
• 2018

Purpose: We performed quantitative and qualitative analysis of typical periodontal bacteria using real time PCR method to investigate the microbiological difference according to the severity of peri-implant disease in Koreans. Materials and Methods: Total of 60 implants were divided into three groups (healthy group, peri-implant mucositis group, peri-implantitis group) through periapical radiographs and clinical indices. The evaluated clinical parameters were pocket depth, plaque index, suppuration and bleeding on probing. Using a sterilized curette instrument, microbial samples were collected from the subgingival plaque and real-time PCR was performed on five periodontal bacteria. The relative expression levels of microorganisms were compared by comparative delta-CT method. Results: The relative expression levels of E. corrodens and T. denticola were significantly higher in the peri-implantitis group (P < 0.017). On the other hand, the relative expression level of F. nucleatum and P. gingivalis was relatively high in the healthy implant group regardless of the severity of disease. P. intermedia was significantly lower in the healthy implant group (P < 0.017). Conclusion: Periodontal bacteria were detected in Koreans with peri-implant diseases, but there was no microbiological distribution similar to periodontitis.

• Journal of Apiculture
• /
• v.32 no.2
• /
• pp.119-131
• /
• 2017

For the Rapid detection of small hive beetle (SHB; Aethina tumida) and for the mass-survey against SHB invasion, SHB-specific ultra-rapid PCR system was developed. Three different pairs of Aethina tumida-specific primers were deduced from cytochrome oxidase subunit I (COI) gene in mitochondrial DNA of SHB. Using optimized SHB-specific ultra-rapid PCR, $2.1{\times}10^1$ molecules of COI gene belonged to SHB could be detected specifically and quantitatively within 18 minutes 40 seconds. For the purpose of the application in apiary field, a DNA extraction method from bee debris was separatedly developed. When $10^5$ SHB-specific COI molecules (1/1000 body of SHB larvae) are existed in 1g of bee debris, it could be verified inner 10 minutes as qualitative and quantitative manner. SHB-specific ultra-rapid PCR we proposed would be expected to apply widely, either in apiary field or laboratory, for the rapid detections and the control against SHB-invasion.

• Journal of Korean Academy of Dental Administration
• /
• v.6 no.1
• /
• pp.11-18
• /
• 2018

The aim of this study was to evaluate the competency of the Easyperio test, a genetic test method based on real time PCR for the detection of bacteria that cause dental caries and periodontal disease. To verify the validity of this text, various dental health evaluations were administered to 33 boys between the ages of 12 to 14, as this age group commonly experiences dental caries. These evaluations included a dental caries experience survey, a first molar health evaluation, the Dentocult Streptococcus mutans (SM) strip mutans, the Dentocult Lactobacillus spp (LB) test, and the Easyperio test. The correlation coefficients between the level of the Dentocult SM strip mutans and the dental caries experience were DT (R=0.570, p=0.001), DMFT (R=0.376, p=0.031), and first molar health (R=-0.395, p=0.023). The correlation coefficients between the amount of SM in the Easyperio test and dental caries experience were DT (R=0.528, p=0.002), DMFT (R=0.369, p=0.035), and first molar health (R=-0.426, p=0.013). The correlation coefficients between the level of the dentocult SM strip mutans and the SM amounts of the Easyperio test were S.mi (R=0.564 p=0.001) and S.mu (R=0.621, p=0.002). The correlation coefficients between the level of the Dentocult LB test and the SM amount of Easyperio test was S.mi (R=0.495, p=0.003). In conclusion, Easyperio test may be an easy and effective method for the differentiation and diagnosis of dental caries through quantitative and qualitative analysis of oral bacteria.

• Journal of Korean Society of Environmental Engineers
• /
• v.31 no.10
• /
• pp.919-926
• /
• 2009

Enrichment of anaerobic ammonium oxidation (ANAMMOX) bacteria is the essential step for operating full-scale ANAMMOX bioreactor because adding a significant amount of seeding sludge is required to stabilize the ANAMMOX reactor. In this study, the enrichment of ANAMMOX bacteria from an activated sludge using sequencing batch reactor was conducted and verified by analyzing changes in the microbial community structure. ANAMMOX bacteria were successfully enriched for 70 days and the substrate removal efficiencies showed 98.5% and 90.7% for $NH_4\;^+$ and $NO_2\;^-$ in the activity test, respectively. The phylogenetic trees of Planctomycetes phylum showed that the diverse microbial community structure of an activated sludge was remarkably simplified after the enrichment. All 36 clones, obtained after the enrichment, were affiliated with ANAMMOX bacteria of Candidatus Brocadia (36%) and Candidatus Anammoxoglobus (64%) genera. The quantification using real-time quantitative PCR (RTQ-PCR) revea ed that the 16S rDNA concentration of ANAMMOX bacteria was 74.8% compared to the granular ANAMMOX sludge obtained from an upflow ANAMMOX sludge bed reactor which had been operated for more than one year. The results of molecular analysis supported that the enriched sludge could be used as a seeding sludge for a full-scale ANAMMOX bioreactor.