• Journal of Veterinary Science
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• v.22 no.3
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• pp.40.1-40.12
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• 2021

Background: The microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen. Objectives: The present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees. Methods: A procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration. Results: UR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 10⁴ spores/bee, and the stable detection level was ≥ 2.40 × 10⁵ spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 10⁴ copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min. Conclusions: UR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.

• The Plant Pathology Journal
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• v.19 no.4
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• pp.205-209
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• 2003

Dispersal of citrus bacterial canker caused by Xanthomonas axonopodis pv. citri on Unshiu orange was investigated in naturally infested nursery plot at Seogwipo in Jeju island, Korea. Based on phage detection, over 2% of the bacterial pathogen over-wintered in canker lesions and started to multiply in late May. However, symptoms were first observed 1 month after the phage detection. The disease dispersed non-directionally to nearby plants possibly because of indirect dissemination of the bacterium by rain splashes. The disease increased from late June to late August and decreased thereafter. Population of phage increased constantly, however, disease occurrence somewhat fluctuated due to environmental factors. Disease incidence and severity were correlated with rainfall with wind that occurred 14-32 days earlier from late May to late August.

• The Plant Pathology Journal
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• v.36 no.5
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• pp.459-467
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• 2020

Areca palm yellow leaf (AYL) disease caused by the 16SrI group phytoplasma is a serious threat to the development of the Areca palm industry in China. The 16S rRNA gene sequence was utilized to establish a rapid and efficient detection system efficient for the 16SrI-B subgroup AYL phytoplasma in China by loop-mediated isothermal amplification (LAMP). The results showed that two sets of LAMP detection primers, 16SrDNA-2 and 16SrDNA-3, were efficient for 16SrI-B subgroup AYL phytoplasma in China, with positive results appearing under reaction conditions of 64℃ for 40 min. The lowest detection limit for the two LAMP detection assays was the same at 200 ag/μl, namely approximately 53 copies/μl of the target fragments. Phytoplasma was detected in all AYL disease samples from Baoting, Tunchang, and Wanning counties in Hainan province using the two sets of LAMP primers 16SrDNA-2 and 16SrDNA-3, whereas no phytoplasma was detected in the negative control. The LAMP method established in this study with comparatively high sensitivity and stability, provides reliable results that could be visually detected, making it suitable for application and research in rapid diagnosis of AYL disease, detection of seedlings with the pathogen and breeding of disease-resistant Areca palm varieties.

• Korean Journal of Veterinary Research
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• v.57 no.1
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• pp.31-36
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• 2017

Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT ($PRNT_{90}$). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and $PRNT_{90}$ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and $PRNT_{90}$ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.

• Korean Journal of Veterinary Research
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• v.63 no.2
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• pp.19.1-19.6
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• 2023

Rapid immunochromatography test (RICT) kits are commonly used for the diagnosis of canine parvovirus (CPV) because of their rapid turnaround time, simplicity, and ease of use. However, the potential for cross-reactivity and low sensitivity can yield false-positive or false-negative results. There are 4 genotypes of CPV. Therefore, evaluating the performance and reliability of RICT kits for CPV detection is essential to ensure accurate diagnosis for appropriate treatment. In this study, we evaluated the performance of commercial RICT kits in the diagnosis of all CPV genotypes. The cross-reactivity of 6 commercial RICT kits was evaluated using 8 dog-related viruses and 4 bacterial strains. The limit of detection (LOD) was measured for the 4 genotypes of CPV and feline panleukopenia virus. The tested kits showed no cross-reactivity with the 8 dog-related viruses or 4 bacteria. Most RICT kits showed strong positive results for CPV-2 variants (CPV-2a, CPV-2b, and CPV-2c). However, the 2 kits produced negative results for CPV-2 or CPV-2b at a titer of 10⁵ FAID₅₀/mL, which may result in inaccurate diagnoses. Therefore, some kits need to improve their LOD by increasing their binding efficiency to detect all CPV genotypes.

• Korean Journal of Plant Resources
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• v.23 no.3
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• pp.233-241
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• 2010

In vitro elimination of Sweet potato leaf curl virus (SPLCV) from infected sweet potato is difficult due to low number of virus-free plants obtained from meristem tip culture and long growth period required for the virus detection. In this study, efficient production of the SPLCV-free sweet potato by in vitro therapy coupled with a PCR assay for virus detection was investigated. Infected shoots cultured on Murashige and Skoog medium were treated at three different temperatures for 7 weeks followed by meristem tip culture on the medium with or without ribavirin at 50 mg/L. The regenerated plantlets were tested for virus infection by a PCR assay. The results showed that the both heat- and cold-treatments, and addition of the ribavirin did not have significant effect on efficiency of the virus elimination. The meristem size, however, greatly affected the survival rate. Meristems sized over 0.4 mm survived better than smaller ones (0.2-0.3 mm). The PCR assay was approved to be a rapid, sensitive and reliable for the SPLCV detection in regenerated plantlets. Therefore, combination of cultivating meristem tips sized 0.4-0.5 mm on the medium at $22^{\circ}C$ without ribavirin and detection of SPLCV in the regenerated plantlets by the PCR assay was an efficient system for the SPLCV elimination from infected sweet potato.

• Korean Journal Plant Pathology
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• v.11 no.2
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• pp.101-106
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• 1995

Deep blue fluorescence of resorcin blue-stained callose was observed only in the potato leafroll virus (PLRV)-infected potato plants, but not in other potato viruses investigated. The plant sections stained with aniline blue showed non-specific fluorescence regardless of PLRV infection, which means that aniline blue is not suitable for the staining of callose induces by PLRV infection. The fluorescence of resorcin blue-stained callose was more easily detectable than autofluorescence by a direct fluorescence detection method because of its deep blue color. The lateral branch of lower leaves was turned out to be the best material for fluorescence observation of all plant parts tested. In comparison of diagnostic efficacy of this technique to enzyme-linked immunosorbent assay (ELISA), PLRV infected potato plants showed corresponding increment of the fluorescence of resorcin blue stained callose as absorption values in ELISA increased. In the future, the criteria measuring the fluorescence objectively are thought to be determined for the practical application to the diagnosis of potato leafroll disease.

• Research in Plant Disease
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• v.17 no.2
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• pp.216-221
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• 2011

A representative isolate KU53 of the predominant Penicillium species was obtained from rice samples from rice processing complexes of National Agricultural Cooperative Federation in Korea. In this study, isolate KU53 was identified by its morphological and molecular characteristics. The macro- and microscopic characteristics of isolate KU53 were compared with the P. fellutanum reference isolate KCTC16913 on different media; isolate KU53 was generally identical to those of the reference isolate KCTC16913. In a molecular-based identification, the ${\beta}$-tubulin and translation elongation factor 1-alpha sequences of isolate KU53 was most closely related to those of P. fellutanum. Thus, isolate KU53 from stored rice could be identified as P. fellutanum, some isolates of which are known to produce mycotoxin-related metabolites. To our knowledge, this is the first detection of P. fellutanum from stored rice in Korea.

• The Plant Pathology Journal
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• v.36 no.3
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• pp.204-217
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• 2020

In nature, plants are always under the threat of pests and diseases. Pathogenic bacteria are one of the major pathogen types to cause diseases in diverse plants, resulting in negative effects on plant growth and crop yield. Chemical bactericides and antibiotics have been used as major approaches for controlling bacterial plant diseases in the field or greenhouse. However, the appearance of resistant bacteria to common antibiotics and bactericides as well as their potential negative effects on environment and human health demands bacteriologists to develop alternative control agents. Bacteriophages, the viruses that can infect and kill only target bacteria very specifically, have been demonstrated as potential agents, which may have no negative effects on environment and human health. Many bacteriophages have been isolated against diverse plant-pathogenic bacteria, and many studies have shown to efficiently manage the disease development in both controlled and open conditions such as greenhouse and field. Moreover, the specificity of bacteriophages to certain bacterial species has been applied to develop detection tools for the diagnosis of plant-pathogenic bacteria. In this paper, we summarize the promising results from greenhouse or field experiments with bacteriophages to manage diseases caused by plant-pathogenic bacteria. In addition, we summarize the usage of bacteriophages for the specific detection of plant-pathogenic bacteria.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.121.1-121
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• 2003

Dispersal of Xanthomonas axonopodis pv. citri, causing citrus bacterial canker disease on Unshiu orange was investigated at previously infested plots at Seogwipo in Jeju island of Korea. The bacterial pathogen overwintered in lesions started to multiply at tate May, and disease firstly observed one month after detection of phage from lesions. The disease gradually increased, however, it dispersed non-directionally to nearby plants from inoculum sources. Diseased plants were aggregated to form a cluster throughout the experiment. Population dynamics of phage on symtomless leaf surface and the disease severity were compared in the nursery, Increase of phage population on symptomless leaf surface preceded one month to that of the disease severity Population of phage increased constantly from late July to October, however, the disease severity decreased from late August to late October. It was assumed that the decrease of disease severity might be due to disease-induced defoliation.