• Title/Summary/Keyword: Gene testing

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Molecular biological analysis of Bt-transgenic (Bt-9) rice and its effect on Daphnia magna feeding

  • Oh, Sung-Dug;Yun, Doh-Won;Chang, Ancheol;Lee, Yu-jin;Lim, Myung-Ho
    • Korean Journal of Agricultural Science
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    • v.46 no.1
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    • pp.113-124
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    • 2019
  • Insect-resistant transgenic (Bt-9) rice was generated by inserting mCry1Ac1, a modified gene from the soil bacterium Bacillus thuringiensis, into the genome of a conventional variety of rice (Ilmi). With regard to potential problems such as safety, an evaluation of non-target organisms is necessary as an essential element of an environmental risk assessment of genetically modified (GM) crops. We studied the effects of the Bt-9 rice on the survival of cantor Daphnia magna, a commonly used model organism in ecotoxicological studies. D. magna fed on the Bt-transgenic rice (Bt-9) and its near non-GM counterparts (Ilmi) grown in the same environment (a 100% ground rice suspension). The Bt-9 rice was confirmed to have the inserted T-DNA and protein expression evident by the PCR and ELISA analyses. The feeding study showed a similar cumulative immobility and abnormal response of the Daphnia magna between the Bt-9 rice and Ilmi. Additionally, the 48 h-EC50 values of the Bt-9 and Ilmi rice were 4,400 mg/L (95% confidence limits: 3861.01 - 5015.01 mg/L) and 5,564 mg/L (95% confidence limits: 4780.03 - 6476.93 mg/L), respectively. The rice NOEC (No observed effect concentration) value for D. magna was suggested to be 1,620 mg/L. We conclude that the tested Bt-9 and Ilmi have a similar cumulative immobility for D. magna, a widely used model organism, and the growth of Bt-9 did not affect non-target insects.

Genotype-phenotype correlations in pediatric patients with myotonic dystrophy type 1

  • Kim, Hyeong Jung;Na, Ji-Hoon;Lee, Young-Mock
    • Clinical and Experimental Pediatrics
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    • v.62 no.2
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    • pp.55-61
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    • 2019
  • Purpose: Myotonic dystrophy, also known as dystrophia myotonica (DM), is an autosomal dominant disorder with 2 genetically distinct forms. DM type 1 (DM1) is the more common form and is caused by abnormal expansion of cytosine/thymine/guanine (CTG) repeats in the DM protein kinase (DMPK ) gene. Our study aimed to determine whether the age of onset is correlated with CTG repeat length in a population of pediatric patients with DM1. Methods: We retrospectively identified 30 pediatric patients with DM1 that underwent DMPK testing, of which the clinical data of 17 was sufficient. The cohort was divided into 2 subgroups based on the clinical phenotype (congenital-onset vs. late-onset) and number of CTG repeats (<1,000 vs. ${\geq}1,000$). Results: We found no significant difference between the age of onset and CTG repeat length in our pediatric patient population. Based on clinical subgrouping, we found that the congenital-onset subgroup was statistically different with respect to several variables, including prematurity, rate of admission to neonatal intensive care unit, need for respiratory support at birth, hypotonia, dysphagia, ventilator dependence, and functional status on last visit, compared to the late-onset subgroup. Based on genetic subgrouping, we found a single variable (poor feeding in neonate) that was significantly different in the large CTG subgroup than that in the small CTG subgroup. Conclusion: Clinical variables exhibiting statistically significant differences between the subgroups should be focused on prognosis and designing tailored management approaches for the patients; our findings will contribute to achieve this important goal for treating patients with DM1.

Development and validation of a PCR method to discriminate between Branchiostegus japonicus and Branchiostegus albus (옥돔과 옥두어 판별을 위한 PCR 검사법 개발과 검증)

  • Kim, Na-Ye-Seul;Yang, Ji-Young;Kim, Jung-Beom
    • Korean Journal of Food Science and Technology
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    • v.51 no.3
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    • pp.295-299
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    • 2019
  • We developed and validated species-specific primers for Branchiostegus japonicus and Branchiostegus albus to prevent the sale of B. albus as B. japonicus. Primers for B. japonicus and B. albus were designed against the cytochrome b gene. Multiplex PCR showed a 288 bp amplicon for B. japonicus, a 159 bp amplicon for B. albus, and a 502 bp amplicon for the internal control. The PCR product bands for B. japonicus, B. albus, and the internal control were present at 1 ng each. The specificity and sensitivity of the primers developed in this study were validated by testing 38 B. japonicus strains and 13 B. albus strains. Using this monitoring method, fake fish did not appear due to the agreement between the experimental results and the species. Therefore, the developed multiplex PCR method was suitable for differentiating B. japonicus and B. albus.

Current status of food safety detection methods for Smart-HACCP system (스마트-해섭(Smart-HACCP) 적용을 위한 식품안전 검시기술 동향)

  • Lim, Min-Cheol;Woo, Min-Ah;Choi, Sung-Wook
    • Food Science and Industry
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    • v.54 no.4
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    • pp.293-300
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    • 2021
  • Food safety accidents have been increasing by 2% over 5,000 cases every year since 2009. Most people know that the best method to prevent food safety accidents is a quick inspection, but there is a lack of inspection technology that can be used at the non-analytic level to food production and distribution sites. Among the recent on-site diagnostic technologies, the methods for testing gene-based food poisoning bacteria were introduced with the STA technology, which can range from sample to detection. If food safety information can be generated without forgery by directly inspecting food hazard factors by remote, unmanned, not human, pollution sources can be managed by predicting risks more accurately from current big-data and artificial intelligence technology. Since this information processing can be used on smartphones using the current cloud technology, it is judged that it can be used for food safety to small food businesses or catering services.

Genetic Variation of Flower Production in Breeding Seedling Seed Orchards of Quercus acuta and Q. glauca

  • Jeon, Koeun;Ro, Hee Seung;Kim, Ye-Ji;Gu, Da-Eun;Park, Ji-Min;Ryu, Sungryul;Kang, Kyu-Suk
    • Journal of Forest and Environmental Science
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    • v.38 no.2
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    • pp.102-109
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    • 2022
  • This study was conducted to test the significant difference of fertility variation among families and to select superior families for acorn production in the breeding seedling seed orchards (BSSOs) of Quercus acuta and Quercus glauca. The seed orchards were located in Jeju island and established by seedlings raised from selected parents for genetic testing in 2006. In the spring of 2021, the numbers of female and male flower were counted from 5 to 10 individuals per family in the BSSOs. To test statistical significance of which parameter is not satisfied through the normality test, we used a nonparametric analysis. Correlation analysis was performed to quantify the association between female and male flower production. As the results, the significant difference of flower production among families was found in both seed orchards. The averages of female flower production were 65.3 and 181.9 in Q. acuta and Q. glauca. The positive Spearman's rank correlation was existed between male and female flower production. Broad-sense heritability on female and male flower production were 0.191 and 0.147 in Q. acuta, and 0.285 and 0.068 in Q. glauca, respectively. Sexual asymmetry (e.g., maleness index) between female and male, and contribution variation among families (e.g., parental balance) were analyzed to find reasonable alternatives in the management of seed orchards. Effective population size of seed crops was predicted as a concept of status number. Loss of gene diversity (accumulation of group coancestry) would not be alarming in the BSSOs. Our results would be helpful to select breeding materials for establishing new seed orchards and to supply genetically improved seeds of evergreen oaks, which is one of the backbones of the strategy of carbon sink in the 2050 Carbon Neutrality of Korea Forest Service.

Designing a novel mRNA vaccine against Vibrio harveyi infection in fish: an immunoinformatics approach

  • Islam, Sk Injamamul;Mou, Moslema Jahan;Sanjida, Saloa;Tariq, Muhammad;Nasir, Saad;Mahfuj, Sarower
    • Genomics & Informatics
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    • v.20 no.1
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    • pp.11.1-11.20
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    • 2022
  • Vibrio harveyi belongs to the Vibrio genus that causes vibriosis in marine and aquatic fish species through double-stranded DNA virus replication. In humans, around 12 Vibrio species can cause gastroenteritis (gastrointestinal illness). A large amount of virus particles can be found in the cytoplasm of infected cells, which may cause death. Despite these devastating complications, there is still no cure or vaccine for the virus. As a result, we used an immunoinformatics approach to develop a multi-epitope vaccine against most pathogenic hemolysin gene of V. harveyi. The immunodominant T- and B-cell epitopes were identified using the hemolysin protein. We developed a vaccine employing three possible epitopes: cytotoxic T-lymphocytes, helper T-lymphocytes, and linear B-lymphocyte epitopes, after thorough testing. The vaccine was developed to be antigenic, immunogenic, and non-allergenic, as well as having a better solubility. Molecular dynamics simulation revealed significant structural stiffness and binding stability. In addition, the immunological simulation generated by computer revealed that the vaccination might elicit immune reactions in the actual life after injection. Finally, using Escherichia coli K12 as a model, codon optimization yielded ideal GC content and a higher codon adaptation index value, which was then included in the cloning vector pET2+ (a). Altogether, our experiment implies that the proposed peptide vaccine might be a good option for vibriosis prophylaxis.

Comparative Performance of Line Probe Assay (Version 2) and Xpert MTB/RIF Assay for Early Diagnosis of Rifampicin-Resistant Pulmonary Tuberculosis

  • Yadav, Raj Narayan;Singh, Binit Kumar;Sharma, Rohini;Chaubey, Jigyasa;Sinha, Sanjeev;Jorwal, Pankaj
    • Tuberculosis and Respiratory Diseases
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    • v.84 no.3
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    • pp.237-244
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    • 2021
  • Background: The emergence of drug-resistant tuberculosis (TB), is a major menace to cast off TB worldwide. Line probe assay (LPA; GenoType MTBDRplus ver. 2) and Xpert MTB/RIF assays are two rapid molecular TB detection/diagnostic tests. To compare the performance of LPA and Xpert MTB/RIF assay for early diagnosis of rifampicin-resistant (RR) TB in acid-fast bacillus (AFB) smear-positive and negative sputum samples. Methods: A total 576 presumptive AFB patients were selected and subjected to AFB microscopy, Xpert MTB/RIF assay and recent version of LPA (GenoType MTBDRplus assay version 2) tests directly on sputum samples. Results were compared with phenotypic culture and drug susceptibility testing (DST). DNA sequencing was performed with rpoB gene for samples with discordant rifampicin susceptibility results. Results: Among culture-positive samples, Xpert MTB/RIF assay detected Mycobacterium tuberculosis (Mtb) in 97.3% (364/374) of AFB smear-positive samples and 76.5% (13/17) among smear-negative samples, and the corresponding values for LPA test (valid results with Mtb control band) were 97.9% (366/374) and 58.8% (10/17), respectively. For detection of RR among Mtb positive molecular results, the sensitivity of Xpert MTB/RIF assay and LPA (after resolving discordant phenotypic DST results with DNA sequencing) were found to be 96% and 99%, respectively. Whereas, specificity of both test for detecting RR were found to be 99%. Conclusion: We conclude that although Xpert MTB/RIF assay is comparatively superior to LPA in detecting Mtb among AFB smear-negative pulmonary TB. However, both tests are equally efficient in early diagnosis of AFB smear-positive presumptive RR-TB patients.

Development of reverse transcription loop-mediated isothermal amplification assays for point-of-care testing of avian influenza virus subtype H5 and H9

  • Zhang, Songzi;Shin, Juyoun;Shin, Sun;Chung, Yeun-Jun
    • Genomics & Informatics
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    • v.18 no.4
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    • pp.40.1-40.8
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    • 2020
  • Avian influenza (AIV) outbreaks can induce fatal human pulmonary infections in addition to economic losses to the poultry industry. In this study, we aimed to develop a rapid and sensitive point-of-care AIV test using loop-mediated isothermal amplification (LAMP) technology. We designed three sets of reverse transcription LAMP (RT-LAMP) primers targeting the matrix (M) and hemagglutinin (HA) genes of the H5 and H9 subtypes. RT-LAMP targeting the universal M gene was designed to screen for the presence of AIV and RT-LAMP assays targeting H5-HA and H9-HA were designed to discriminate between the H5 and H9 subtypes. All three RT-LAMP assays showed specific amplification results without nonspecific reactions. In terms of sensitivity, the detection limits of our RT-LAMP assays were 100 to 1,000 RNA copies per reaction, which were 10 times more sensitive than the detection limits of the reference reverse-transcription polymerase chain reaction (RT-PCR) (1,000 to 10,000 RNA copies per reaction). The reaction time of our RT-LAMP assays was less than 30 min, which was approximately four times quicker than that of conventional RT-PCR. Altogether, these assays successfully detected the existence of AIV and discriminated between the H5 or H9 subtypes with higher sensitivity and less time than the conventional RT-PCR assay.

Studies on Alkali Digestibility of Soybean Seeds and Its Inheritance (콩에 있어서 알칼리 붕괴도와 그의 유전에 관한 연구)

  • Lee, H.S.;Park, E.H.;Lim, B.K.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.29 no.2
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    • pp.179-185
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    • 1984
  • These studies were carried out to evaluate the feasibility of using Alkali Digestibility Value (ADV) of soybean seeds as a selection criterion for cooking quality of soybean grain with rice, and to find out an optimal testing condition, varietal differences in ADV and the effect of cultural environment on the ADV, and its mode of inheritance. The ADV test was achieved succesfully by soaking the soybean grain for 24 hours in 25 to 30% KOH solution at 25 to $30^{\circ}C$. Varietal differences in the ADV were conspicuous, but cultural conditions affected the ADV little. All collected lines for cooking with rice showed high ADV and all the small grain cultivars introduced from abroad had low ADV, while recommended cultivars showed great variation in ADV. The characteristic of low ADV in soybean grains was controlled by single dominant gene.

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Evaluation of Loop Mediated Isothermal Amplification Based Methods for the Detection of African Swine Fever Virus from Food Waste

  • Siwon Lee;Junhwa Kwon;Su Hyang Kim;Jin-Ho Kim;Jaewon Jung;Kyung-Jin Lee;Ji-Yeon Park;Taek-Kyun Choi;Jun-Gu Kang;Tae Uk Han
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.334-339
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    • 2022
  • African swine fever virus (ASFV) is a highly contagious and lethal pathogen that poses a threat to the global pork industry. The World Organization for Animal Health (WOAH) has placed strict surveillance measures for ASFV. The possibility of long-term survival of ASFV in raw meat or undercooked pork has been reported. Accordingly, the problem of secondary infection in food waste from households or waste disposal facilities has emerged, raising the need for ASFV monitoring of food waste. However, most of the previously reported ASFV gene detection methods are focused on clinical monitoring of pigs. There are very few cases in which their application in waste has been verified. Since ASFV diagnosis requires rapid monitoring and immediate action, loop-mediated isothermal amplification (LAMP) may be suitable, but this requires conformity assessment for LAMP to be used as a diagnostic technique. In this study, six LAMP methods were evaluated, and two methods (kit and manual) were recommended for use in diagnosing ASFV in food waste.