• Title/Summary/Keyword: Assay sensitivity

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DEVELOPMENT OF LEAD SLOWING DOWN SPECTROMETER FOR ISOTOPIC FISSILE ASSAY

  • Lee, YongDeok;Park, Chang Je;Ahn, Sang Joon;Kim, Ho-Dong
    • Nuclear Engineering and Technology
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    • v.46 no.6
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    • pp.837-846
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    • 2014
  • A lead slowing down spectrometer (LSDS) is under development for analysis of isotopic fissile material contents in pyro-processed material, or spent fuel. Many current commercial fissile assay technologies have a limitation in accurate and direct assay of fissile content. However, LSDS is very sensitive in distinguishing fissile fission signals from each isotope. A neutron spectrum analysis was conducted in the spectrometer and the energy resolution was investigated from 0.1eV to 100keV. The spectrum was well shaped in the slowing down energy. The resolution was enough to obtain each fissile from 0.2eV to 1keV. The detector existence in the lead will disturb the source neutron spectrum. It causes a change in resolution and peak amplitude. The intense source neutron production was designed for ~E12 n's/sec to overcome spent fuel background. The detection sensitivity of U238 and Th232 fission chamber was investigated. The first and second layer detectors increase detection efficiency. Thorium also has a threshold property to detect the fast fission neutrons from fissile fission. However, the detection of Th232 is about 76% of that of U238. A linear detection model was set up over the slowing down neutron energy to obtain each fissile material content. The isotopic fissile assay using LSDS is applicable for the optimum design of spent fuel storage to maximize burnup credit and quality assurance of the recycled nuclear material for safety and economics. LSDS technology will contribute to the transparency and credibility of pyro-process using spent fuel, as internationally demanded.

Loop-mediated isothermal amplification assay for differentiation of Mycobacterium bovis and M. tuberculosis (Mycobacterium bovis와 M. tuberculosis 감별을 위한 등온증폭법)

  • Koh, Ba-Ra-Da;Kim, Jae-Myung;Sung, Chang-Min;Ji, Tae-Kyung;Na, Ho-Myung;Park, Seong-Do;Kim, Yong-Hwan;Kim, Eun-Sun
    • Korean Journal of Veterinary Service
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    • v.36 no.2
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    • pp.79-86
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    • 2013
  • Mycobacterium (M.) bovis, a member of the M. tuberculosis complex (MTC), is a re-emerging, zoonotic agent of bovine tuberculosis whose prevalence probably depends on variations in direct exposure to cattle and ingestion of raw milk. Accurate species differentiation of M. bovis and M. tuberculosis is needed to distinguish between human and zoonotic tuberculosis. This study successfully developed a loop-mediated isothermal amplification (LAMP) assay for rapid detection and differentiation of M. bovis and M. tuberculosis, however showed negative reactions in eight non-tuberculous mycobacteria (NTM) samples and ten other bacterial species. Sensitivity of this assay for detection of genomic M. bovis DNA was 10 $fg/{\mu}l$. And this assay successfully detected M. bovis in bovine clinical specimens. In conclusion, the LAMP assay is a simple and powerful tool for rapid detection of M. bovis in both pure bacterial culture and in clinical samples.

Detection of Rifampicin- and Isoniazid-Resistant Mycobacterium tuberculosis Using the Quantamatrix Multiplexed Assay Platform System

  • Wang, Hye-young;Uh, Young;Kim, Seoyong;Cho, Eunjin;Lee, Jong Seok;Lee, Hyeyoung
    • Annals of Laboratory Medicine
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    • v.38 no.6
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    • pp.569-577
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    • 2018
  • Background: The increasing prevalence of drug-resistant tuberculosis (TB) infection represents a global public health emergency. We evaluated the usefulness of a newly developed multiplexed, bead-based bioassay (Quantamatrix Multiplexed Assay Platform [QMAP], QuantaMatrix, Seoul, Korea) to rapidly identify the Mycobacterium tuberculosis complex (MTBC) and detect rifampicin (RIF) and isoniazid (INH) resistance-associated mutations. Methods: A total of 200 clinical isolates from respiratory samples were used. Phenotypic anti-TB drug susceptibility testing (DST) results were compared with those of the QMAP system, reverse blot hybridization (REBA) MTB-MDR assay, and gene sequencing analysis. Results: Compared with the phenotypic DST results, the sensitivity and specificity of the QMAP system were 96.4% (106/110; 95% confidence interval [CI] 0.9072-0.9888) and 80.0% (72/90; 95% CI 0.7052-0.8705), respectively, for RIF resistance and 75.0% (108/144; 95% CI 0.6731-0.8139) and 96.4% (54/56; 95% CI 0.8718-0.9972), respectively, for INH resistance. The agreement rates between the QMAP system and REBA MTB-MDR assay for RIF and INH resistance detection were 97.6% (121/124; 95% CI 0.9282-0.9949) and 99.1% (109/110; 95% CI 0.9453-1.0000), respectively. Comparison between the QMAP system and gene sequencing analysis showed an overall agreement of 100% for RIF resistance (110/110; 95% CI 0.9711-1.0000) and INH resistance (124/124; 95% CI 0.9743-1.0000). Conclusions: The QMAP system may serve as a useful screening method for identifying and accurately discriminating MTBC from non-tuberculous mycobacteria, as well as determining RIF- and INH-resistant MTB strains.

Preliminary Application of Molecular Monitoring of the Pacific Herring (Clupea pallasii) Based on Real-time PCR Assay Utilization on Environmental Water Samples

  • Kim, Keun-Yong;Heo, Jung Soo;Moon, Seong Yong;Kim, Keun-Sik;Choi, Jung-Hwa;Yoo, Joon-Taek
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.209-220
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    • 2021
  • Pacific herring, Clupea pallasii, a keystone species with significant ecological and commercial importance, is declining globally throughout much of its range. While traditional fishing equipment methods remain limited, new sensitive and rapid detection methods should be developed to monitor fisheries resources. To monitor the presence and quantity of C. pallasii from environmental DNA (eDNA) extracted from seawater samples, a pair of primers and a TaqMan® probe specific to this fish based on mitochondrial cytochrome b (COB) sequences were designed for the real-time PCR (qPCR) assay. The combination of our molecular markers showed high specificity in the qPCR assay, which affirmed the success of presenting a positive signal only in the C. pallasii specimens. The markers also showed a high sensitivity for detecting C. pallasii genomic DNA in the range of 1 pg~100 ng rxn-1 and its DNA plasmid containing COB amplicon in the range of 1~100,000copies rxn-1, which produced linear standard calibration curves (r2=0.99). We performed a qPCR assay for environmental water samples obtained from 29 sampling stations in the southeastern coastal regions of South Korea using molecular markers. The assay successfully detected the C. pallasii eDNA from 14 stations (48.2%), with the highest mean concentration in Jinhae Bay with a value of 76.09±18.39 pg L-1 (246.20±58.58 copies L-1). Our preliminary application of molecular monitoring of C. pallasii will provide essential information for efficient ecological control and management of this valuable fisheries resource.

Development of a Novel Multiple Cross-Linking Spiral Amplification for Rapid and Sensitive Detection of HPV16 DNA

  • Zhang, Donghong;Liu, Dongliang;Liu, Bing;Ma, Xiulan
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.610-620
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    • 2021
  • There has been increasing interest in the head and neck squamous cell carcinoma (HNSCC) that is caused by high-risk human papillomavirus (HR-HPV) and has posed a significant challenge to Otolaryngologists. A rapid, sensitive, and reliable method is required for the detection of HR-HPV in clinical specimens to prevent and treat HPV-induced diseases. In this study, a multiple cross-linking spiral amplification (MCLSA) assay was developed for the visual detection of HPV-16. In the MCLSA assay, samples were incubated under optimized conditions at 62℃ for 45 min, and after mixing with the SYBR Green I (SGI) dye, the positive amplicons showed bright green fluorescence while the negative amplicons exhibited no obvious change. The specificity test revealed that the developed MCLSA technique had high specificity and could effectively distinguish all five HPV-16 strains from other pathogenic microorganisms. In terms of analytical sensitivity, the limit of detection (LoD) of MCLSA assay was approximately 5.4 × 101 copies/tube, which was 10-fold more sensitive than loop-mediated isothermal amplification (LAMP) and RT-PCR. The detection results of laryngeal cancer specimens collected from 46 patients with suspected HPV infection in the Liaoning region demonstrated that the positive detection rates of MCLSA and hybridized capture 2 kit were 32.61% (15/46). The true positive rate of the MCLSA assay was higher than that of RT-PCR (100% vs. 93.33%) and LAMP (100% vs. 86.67%). Therefore, the MCLSA assay developed in the present study could be a potentially useful tool for the point-of-care (PoC) diagnosis of HR-HPV, especially in resource-limited countries.

Development of oligonucleotide microarray system for differential diagnosis of enteric viruses in diarrheic fecal samples in pigs

  • Park, Nam-Yong;Kim, Yong-Hwan;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.30 no.4
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    • pp.489-496
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    • 2007
  • An oligonucleotide microarray system was developed for the simultaneous detection of porcine epidemic diarrhea virus, transmissible gastroenteritis virus, porcine enteric calicivirus, porcine group A and C rotavirus. RNAs of the reference viruses and porcine diarrhea samples were extracted and amplified using one-step multiplex RT-PCR in the presence of cyanine 5-dCTP and hybridized on the microarray chip that spotted the virus-specific oligonucleotides. This system were approximately 10-to 100-fold higher in sensitivity than conventional RT-PCR, and the assay time was less than 3 hours. The relative sensitivity and specificity were 92% and 72.2%, respectively, based on 102 porcine diarrhea samples using RT-PCR as gold standard. These results suggested that the oligonucleotide microarray system in this study be probably more reliable and reproducible means for detecting porcine enteric viruses and that it could be of substantial use in routine diagnostic laboratories.

Enzyme Immunoassay for Rapid Detection of the Fungicide Iprovalicarb Residues (살균제 Iprovalicarb 잔류물의 신속한 검출을 위한 효소면역분석법)

  • Cho, H.K.;Kyung, K.S.;Lee, E.Y.
    • Journal of Biosystems Engineering
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    • v.31 no.6 s.119
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    • pp.535-540
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    • 2006
  • For a biosensor development, an enzyme-linked immunosorbent assay (ELISA) of the fungicide iprovalicarb was developed by minimizing the processing time. The time for whole incubation process was reduced from 135 minutes to 15 minutes. The concentration of antibody was varied to improve sensitivity. The total processing time was reduced from 2.5 hours to 20 minutes, the final sensitivity ($IC_{50}$ value) of 7.93 ng/mL and the lowest detection limit of 0.045 ng/mL were obtained. This ELISA was applied to potatoes and onions, and the recoveries were in the range of 98.85 $\sim$ 101.20% and 87.97 $\sim$ 102.70%, respectively. Accordingly, this method can be used as basis for a biosensor for rapid monitoring of iprovalicarb residues in crops.

A comparison of RPLA and PCR for detection of enterotoxins in methicillin-resistant Staphylococcus aureus(MRSA) strains isolated in dogs

  • Park, Son-il;Han, Hong-ryul
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.806-810
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    • 1999
  • A multiplex-polymerase chain reaction (PCR) assay was used to detect staphylococcal enterotoxin production by 12 strains of Staphylococcus aureus isolated from clinical specimens. To evaluate the efficacy and/or sensitivity of this method, the results were compared to those obtained with the reversed passive latex agglutination kit (SET-RPLA, Denka Seiken, Japan). Of 10 strains positive by PCR were positive by RPLA but two strains, representing high sensitivity of the former method. Enterotoxin B was the most prevalent by the two methods. The kappa index between the two methods was 0.826, indicating a higher agreement and fully reliable for use. These results would suggest that sensitive, inexpensive, and relatively rapid multiplex-PCR technique may be an effective means for the detection of staphylococcal enterotoxin genes as an alternative to traditional methods such as kits or immunological methods, which depend upon the amount of enterotoxin produced.

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Performance of the xTAG$^{(R)}$ Gastrointestinal Pathogen Panel, a Multiplex Molecular Assay for Simultaneous Detection of Bacterial, Viral, and Parasitic Causes of Infectious Gastroenteritis

  • Claas, Eric C.;Burnham, Carey-Ann D.;Mazzulli, Tony;Templeton, Kate;Topin, Francois
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.1041-1045
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    • 2013
  • The xTAG$^{(R)}$ Gastrointestinal Pathogen Panel (GPP) is a multiplexed molecular test for 15 gastrointestinal pathogens. The sensitivity and specificity of this test were assessed in 901 stool specimens collected from pediatric and adult patients at four clinical sites. A combination of conventional and molecular methods was used as comparator. Sensitivity could be determined for 12 of 15 pathogens and was 94.3% overall. The specificity across all 15 targets was 98.5%. Testing for the pathogen identified was not requested by the physician in 65% of specimens. The simultaneous detection of these 15 pathogens can provide physicians with a more comprehensive assessment of the etiology of diarrheal disease.

Comparison of Two PCR Assays for Trichomonas vaginalis

  • Noh, Chang-Suk;Kim, Sang-Su;Park, Sung-Yul;Moon, Hong-Sang;Hong, Yeonchul;Ryu, Jae-Sook
    • Parasites, Hosts and Diseases
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    • v.57 no.1
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    • pp.27-31
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    • 2019
  • PCR is known to be the most sensitive method for diagnosing Trichomonas vaginalis infections. This study aimed to compare the sensitivity of a PCR assay for trichomoniasis (HY-PCR) developed in Hanyang University with the use of a Seeplex Ace Detection $Kit^{(R)}$, using urine collected from four Korean men with prostatic disease. Overall, HY-PCR was more sensitive than the Seeplex Kit. The use of Chelex 100 is recommended for DNA isolation in order to increase the sensitivity of the PCR test.