• Title/Summary/Keyword: Rapid Diagnosis

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Development of Rapid Diagnostic Kit for Identification of Hanwoo (Korean Native Cattle) Brand Meat by Detecting BIO-TAG

  • Baek, Kyung Hoon;Park, Sung Kwon;Lee, Myung Hoon;Kim, Sung Il;Cho, Soo Hyun;Choi, Chang Bon
    • Food Science of Animal Resources
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    • v.34 no.3
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    • pp.339-345
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    • 2014
  • This study was performed to develop a rapid immuno-assay kit, by using a specific antigen to detect Hanwoo brand meat. We selected a synthetic antigen specific to our target antibody, named BIO-TAG (Tyr-D-Ala-Phe), by utilizing a computer-based analysis and literature review. BIO-TAG tagged with adjuvant was subcutaneously injected in sheep and Hanwoo. The serum and meat juice of the immunized or non-immunized animal were then analyzed, to measure the titer of antibody by ELISA and Western blot. The amount of antibodies against the BIO-TAG increased (p<0.05) in serum by vaccination. Furthermore, meat juice from the immunized Hanwoo showed greater (p<0.05) antibody titer, compared with those from non-immunized groups. To optimze the dilution factor, we performed dot-ELISA, with various combination levels of BIO-TAG. Results from dot-ELISA showed that 2 mg/mL BIO-TAG was sufficient to distinguish the immunized meat from non-immunized groups. These results support our hypothesis that simple immunization of Hanwoo generates a sufficient amount of antibodies to be detectable in the meat juice by means of the immune-assay. Therefore, specific Hanwoo brand meat can be more precisely identified by our rapid diagnostic kit. This technology can deter possible fraud of counterfeit meat brands in the Korean domestic market with ease and rapidity; and offers a new tool that guarantees consumers high quality Hanwoo brand beef.

Probe-based qPCR Assay for Rapid Detection of Predominant Candida glabrata Sequence Type in Korea

  • Bae, Jinyoung;Lee, Kyung Eun;Jin, Hyunwoo
    • Biomedical Science Letters
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    • v.25 no.4
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    • pp.407-416
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    • 2019
  • Recent years have seen an increase in the incidence of candidiasis caused by non-albicans Candida (NAC) species. In fact, C. glabrata is now second only to C. albicans as the most common cause of invasive candidiasis. Therefore, the rapid genotyping specifically for C. glabrata is required for early diagnosis and treatment of candidiasis. A number of genotyping assays have been developed to differentiate C. glabrata sequence types (STs), but they have several limitations. In the previous study, multi-locus sequence typing (MLST) has performed with a total of 101 C. glabrata clinical isolates to analyze the prevalent C. glabrata STs in Korea. A total of 11 different C. glabrata STs were identified and, among them, ST-138 was the most commonly classified. Thus, a novel probe-based quantitative PCR (qPCR) assay was developed and evaluated for rapid and accurate identification of the predominant C. glabrata ST-138 in Korea. Two primer pairs and hybridization probe sets were designed for the amplification of internal transcribed spacer 1 (ITS1) region and TRP1 gene. Analytical sensitivity of the probe-based qPCR assay was 100 ng to 10 pg and 100 ng to 100 pg (per 1 μL), which target ITS1 region and TRP1 gene, respectively. This assay did not react with any other Candida species and bacteria except C. glabrata. Of the 101 clinical isolates, 99 cases (98%) were concordant with MLST results. This novel probe-based qPCR assay proved to be rapid, sensitive, highly specific, reproducible, and cost-effective than other genotyping assay for C. glabrata ST-138 identification.

Rapid, Sensitive, and Specific Detection of Clostridium tetani by Loop-Mediated Isothermal Amplification Assay

  • Jiang, Dongneng;Pu, Xiaoyun;Wu, Jiehong;Li, Meng;Liu, Ping
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.1-6
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    • 2013
  • Tetanus is a specific infectious disease, which is often associated with catastrophic events such as earthquakes, traumas, and war wounds. The obligate anaerobe Clostridium tetani is the pathogen that causes tetanus. Once the infection of tetanus progresses to an advanced stage within the wounds of limbs, the rates of amputation and mortality increase manifold. Therefore, it is necessary to devise a rapid and sensitive point-of-care detection method for C. tetani so as to ensure an early diagnosis and clinical treatment of tetanus. In this study, we developed a detection method for C. tetani using loop-mediated isothermal amplification (LAMP) assay, wherein the C. tetani tetanus toxin gene was used as the target gene. The method was highly specific and sensitive, with a detection limit of 10 colony forming units (CFU)/ml, and allowed quantitative analysis. While detecting C. tetani in clinical samples, it was found that the LAMP results completely agreed with those of the traditional API 20A anaerobic bacteria identification test. As compared with the traditional API test and PCR assay, LAMP detection of C. tetani is simple and rapid, and the results can be identified through naked-eye observation. Therefore, it is an ideal and rapid point-of-care testing method for tetanus.

Loop-mediated Isothermal Amplification assay for Detection of Candidatus Liberibacter Asiaticus, a Causal Agent of Citrus Huanglongbing

  • Choi, Cheol Woo;Hyun, Jae Wook;Hwang, Rok Yeon;Powell, Charles A
    • The Plant Pathology Journal
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    • v.34 no.6
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    • pp.499-505
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    • 2018
  • Huanglongbing (HLB, Citrus greening disease) is one of the most devastating diseases that threaten citrus production worldwide. Although HLB presents systemically, low titer and uneven distribution of these bacteria within infected plants can make reliable detection difficult. It was known loop-mediated isothermal amplification (LAMP) method has the advantages of being highly specific, rapid, efficient, and laborsaving for detection of plant pathogens. We developed a new LAMP method targeting gene contained tandem repeat for more rapid and sensitive detection of Candidatus Liberibacter asiaticus (CLas), putative causal agent of the citrus huanglongbing. This new LAMP method was 10 folds more sensitive than conventional PCR in detecting the HLB pathogen and similar to that of real-time PCR in visual detection assay by adding SYBR Green I to mixture and 1% agarose gel electrophoresis. Positive reactions were achieved in reaction temperature 57, 60 and $62^{\circ}C$ but not $65^{\circ}C$. Although this LAMP method was not more sensitive than real-time PCR, it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Thus, we expect that this LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting the CLas in citrus and can be applied for rapid diagnosis is needed.

Development of a ladder-shape melting temperature isothermal amplification (LMTIA) assay for detection of African swine fever virus (ASFV)

  • Wang, Yongzhen;Wang, Borui;Xu, Dandan;Zhang, Meng;Zhang, Xiaohua;Wang, Deguo
    • Journal of Veterinary Science
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    • v.23 no.4
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    • pp.51.1-51.10
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    • 2022
  • Background: Due to the unavailability of an effective vaccine or antiviral drug against the African swine fever virus (ASFV), rapid diagnosis methods are needed to prevent highly contagious African swine fever. Objectives: The objective of this study was to establish the ladder-shape melting temperature isothermal amplification (LMTIA) assay for the detection of ASFV. Methods: LMTIA primers were designed with the p72 gene of ASFV as the target, and plasmid pUC57 was used to clone the gene. The LMTIA reaction system was optimized with the plasmid as the positive control, and the performance of the LMTIA assay was compared with that of the commercial real-time polymerase chain reaction (PCR) kit in terms of sensitivity and detection rate using 200 serum samples. Results: Our results showed that the LMTIA assay could detect the 104 dilution of DNA extracted from the positive reference serum sample, which was the same as that of the commercial real-time PCR kit. The coincidence rate between the two assays was 100%. Conclusions: The LMTIA assay had high sensitivity, good detection, and simple operation. Thus, it is suitable for facilitating preliminary and cost-effective surveillance for the prevention and control of ASFV.

Diagnostic Accuracy of the Quidel Sofia Rapid Influenza Fluorescent Immunoassay in Patients with Influenza-like Illness: A Systematic Review and Meta-analysis

  • Lee, Jonghoo;Song, Jae-Uk;Kim, Yee Hyung
    • Tuberculosis and Respiratory Diseases
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    • v.84 no.3
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    • pp.226-236
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    • 2021
  • Background: Although the Quidel Sofia rapid influenza fluorescent immunoassay (FIA) is widely used to identify influenza A and B, the diagnostic accuracy of this test remains unclear. Thus, the objective of this study was to determine the diagnostic performance of this test compared to reverse transcriptase-polymerase chain reaction. Methods: A systematic literature search was performed using MEDLINE, EMBASE, and the Cochrane Central Register. Pooled sensitivity, specificity, diagnostic odds ratio (DOR), and a hierarchical summary receiver-operating characteristic curve (HSROC) of this test for identifying influenza A and B were determined using meta-analysis. A sensitivity subgroup analysis was performed to identify potential sources of heterogeneity within selected studies. Results: We identified 17 studies involving 8,334 patients. Pooled sensitivity, specificity, and DOR of the Quidel Sofia rapid influenza FIA for identifying influenza A were 0.78 (95% confidence interval [CI], 0.71-0.83), 0.99 (95% CI, 0.98-0.99), and 251.26 (95% CI, 139.39-452.89), respectively. Pooled sensitivity, specificity, and DOR of this test for identifying influenza B were 0.72 (95% CI, 0.60-0.82), 0.98 (95% CI, 0.96-0.99), and 140.20 (95% CI, 55.92-351.54), respectively. The area under the HSROC for this test for identifying influenza A was similar to that for identifying influenza B. Age was considered a probable source of heterogeneity. Conclusion: Pooled sensitivities of the Quidel Sofia rapid influenza FIA for identifying influenza A and B did not quite meet the target level (≥80%). Thus, caution is needed when interpreting data of this study due to substantial betweenstudy heterogeneity.

Development of Recombinase Polymerase Amplification Combined with Lateral Flow Strips for Rapid Detection of Cowpea Mild Mottle Virus

  • Xinyang Wu;Shuting Chen;Zixin Zhang;Yihan Zhang;Pingmei Li;Xinyi Chen;Miaomiao Liu;Qian Lu;Zhongyi Li;Zhongyan Wei;Pei Xu
    • The Plant Pathology Journal
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    • v.39 no.5
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    • pp.486-493
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    • 2023
  • Cowpea mild mottle virus (CPMMV) is a global plant virus that poses a threat to the production and quality of legume crops. Early and accurate diagnosis is essential for effective managing CPMMV outbreaks. With the advancement in isothermal recombinase polymerase amplification and lateral flow strips technologies, more rapid and sensitive methods have become available for detecting this pathogen. In this study, we have developed a reverse transcription recombinase polymerase amplification combined with lateral flow strips (RT-RPA-LFS) method for the detection of CPMMV, specifically targeting the CPMMV coat protein (CP) gene. The RT-RPA-LFS assay only requires 20 min at 40℃ and demonstrates high specificity. Its detection limit was 10 copies/µl, which is approximately up to 100 times more sensitive than RT-PCR on agarose gel electrophoresis. The developed RT-RPA-LFS method offers a rapid, convenient, and sensitive approach for field detection of CPMMV, which contribute to controlling the spread of the virus.

Clinical Diagnosis and Emotional Behavioral Characteristics Study of Children in a Special Education Class in Korean Elementary School (초등학교 특수학급아동의 임상적 진단 및 감정 행동특성 연구)

  • Lim Myung-Ho;Kang Jin-Kyung;Lee Joo-Hyun;Kim Hyun-Woo
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.17 no.2
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    • pp.114-123
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    • 2006
  • Objectives : The special class has been made, bringing rapid increase quantitatively. The authors carried out the child psychiatric interview and evaluation for 9 special-classed children in Asan city to find out clinical diagnosis and emotional/behavioral characteristics. Methods : The child psychiatrists evaluated special class children by DSM-IV and K-SADS-PL. Tools for the evaluation were Child Behavior Checklist- Korean version, Korean Personality Inventory for Children, Children's Depression Inventory, Abbreviated Conners Parent-Teacher Rating Scale-Revised, State-Trait Anxiety Inventory for Children, Vineland Social Maturity Scale, Wechsler Intelligence Scale for Children-III, and Childhood Autism Rating Scale. Results : Ultimately 53 children, consisting of 35 boys(67.9%) and 18 girls(32.1%), participated, and the average age was $10.5{\pm}1.3$ years old. Their measure of Vineland Social Maturity Scale was $78.7{\pm}20.0$, Childhood Autism Rating Scales was $25.4{\pm}9.0$, Child Depression Inventory was $22.2{\pm}5.2$, State-Trait Anxiety Inventory for Children was $35.2{\pm}8.2/36.5{\pm}6.2$, and Abbreviated Conners Parent-Teacher Rating Scale was $11.0{\pm}4.6$. In the clinical diagnosis evaluation, the prevalence rate of learning disorder was decreased compared to early research, ADHD had been newly appeared and depression disorder and anxiety disorder had been increased. Conclusion : This result suggests that a lot of children in a special class have complex emotional and behavioral problems in addition to educational problems.

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Design of Fuzzy Inference-based Deterioration Diagnosis System through Different Image (차 영상을 통한 퍼지 추론 기반 열화 진단 시스템 설계)

  • Kim, Jong-Bum;Choi, Woo-Yong;Oh, Sung-Kwun;Kim, Young-Il
    • Journal of the Korean Institute of Intelligent Systems
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    • v.25 no.1
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    • pp.57-62
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    • 2015
  • In this paper, we design fuzzy inference-based deterioration diagnosis system through different image for rapid as well as efficient diagnosis of electrical equipments. When the deterioration diagnosis of the electrical equipment starts, abnormal state of assigned area is detected by comparing with the temperature of the first normal state of the area. Deterioration state of detected area is diagnosed by using fuzzy inference algorithm. In the fuzzy inference algorithm, fuzzy rules are defined by If-then form and are described as look-up table. Both temperature and its ensuing variation are used as input variables. While triangular membership function is used for the fuzzy input variables of fuzzy rules, singleton membership function is used for the output variable of fuzzy rules. The final output is calculated by using the center of gravity of fuzzy inference method. Experimental data acquired from individual electrical equipments is used in order to evaluate the output performance of the proposed system.

Frozen Section -Application in the Surgical Pathology- (동결절편법(Frozen Section) -외과병리 영역에서의 적용에 대하여-)

  • Chai, Won-Hee;Lee, Tae-Sook;Hong, Suk-Jae
    • Journal of Yeungnam Medical Science
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    • v.3 no.1
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    • pp.179-183
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    • 1986
  • The frozen section technique is a means of intraoperative pathological diagnosis, and a procedure of great value to the surgeon. This method should be accurate, rapid and reliable. This method serves useful purposes, such as determining the presence of tumor, its type(especially whether it is benign or malignant), the adequacy of a biopsy of a suspected lesion, and the conditions of the surgical margins. But, it bears many disadvantages, the most of which is the danger of incorrect diagnosis. We studied the indications, the limitations, and the accuracy of the frozen section method and the materials studies was total of frozen section during recent 3 years. The overall accuracy of the frozen section diagnosis of 809 cases was 98.1% with 0.5% of false negative, 0% of false positive, 0.5% of incorrect histological diagnosis or grading errors, and 0.9% of deferred cases. The tissues submitted were lymph node, gastrointestinal tract, skin subcutaneous tissues in decreasing oder of frequency. The false positive case is not present, while the false negative cases were 4.

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