• Journal of Microbiology
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• 제44권4호
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• pp.403-408
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• 2006

The purpose of this study was to determine the efficacy of a nucleic acid sequence-based amplification (NASBA) method of detecting noroviruses in artificially and naturally contaminated shellfish. We used 58 fecal samples that tested positive for noroviruses with electron microscopy (EM) to develop an NASBA assay for these viruses. Oligonucleotide primers targeting the polymerase coding region were used to amplify the viral RNA in an isothermal process that resulted in the accumulation of RNA amplicons. These amplicons were detected by hybridization with digoxigenin-labeled oligonucleotide probes that were highly specific for genogroup I (GI) and genogroup II (GII) of noroviruses. The expected band of 327bp appeared in denaturing agarose gel without any nonspecific band. The specific signal for each amplicon was obtained through Northern blotting in many repeats. All fecal samples of which 46(79.3%) belonged to GII and 12(20.6%) belonged to GI were positive for noroviruses by EM and by NASBA. Target RNA concentrations as low as 5pg/ml were detected in fecal specimens using NASBA. When the assay was applied to artificially contaminated shellfish, the sensitivity to nucleic acid was 100pg/1.5g shellfish tissue. The potential use of this assay was also confirmed in naturally contaminated shellfish collected from different ponds in Guangzhou city of China, of which 24 (18.76%) out of 128 samples were positive for noroviruses; of these, 19 (79.6%) belonged to GII and 5 (20.4%) belonged to GI. The NASBA assay provided a more rapid and efficient way of detecting noroviruses in fecal samples and demonstrated its potential for detecting noroviruses in food and environmental samples with high specificity and sensitivity.

• 대한화학회지
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• 제55권2호
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• pp.262-267
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• 2011

형광 검출기를 기반으로 한 변성 고성능 액체 크로마토그래피(DHPLC)분석법은 핵산검출에 유용하게 사용되고 있으며 또한 등온증폭 반응법은 병원성 미생물의 효과적인 검출방법으로 알려져 있다. 이 연구에서는 HBV의 조기 분석 방법으로써 사용되는 등온증폭반응법(LAMP)과 변성고성능 액체 크로마토그래피(DHPLC)의 검출한계와 특이성, 그리고 민감도를 평가하였다. 등온증폭 반응법의 검출 시간이 가장 빠른 장점을 보였으나, 변성 고성능 액체 크로마토그래피 분석법이 등온증폭반응법과 real-time PCR분석법과 비교한 결과, 10배 이상의 높은 민감도를 확인 할 수 있었다. 본 결과로서 B형 간염 바이러스의 진단을 위하여, 빠른 검출법으로써 등온증폭 반응법이 유용하게 쓰일 수 있을 것이며 변성 고성능 액체 크로마토그래피 분석법은 좀 더 낮은 병원균의 감염도 검출할 수 있어, 임상에서 유용하게 사용할 수 있을 것이다.

• Journal of Korean Neurosurgical Society
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• 제50권5호
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• pp.426-433
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• 2011

Objective : The Histoculture Drug Response Assay (HDRA), which measures chemosensitivity using minced tumor tissue on drug-soaked gelfoam, has been expected to overcome the limitations of in vitro chemosensitivity test in part. We analyzed interim results of HDRA in malignant gliomas to see if the test can deserve further clinical trials. Methods : Thirty-three patients with malignant gliomas were operated and their tumor samples were examined for the chemosensitivity to 10 chosen drugs by HDRA. The most sensitive chemotherapy regimen among those pre-established was chosen based on the number of sensitive drugs or total inhibition rate (IR) of the regimen. The response was evaluated by 3 month magnetic resonance image. Results : Among 13 patients who underwent total resection of the tumor, 12 showed no evidence of disease and one patient revealed progression. The response rate in 20 patients with residual tumors was 55% (3 complete and 8 partial responses). HDRA sensitivity at the cut-off value of more than one sensitive drug in the applied regimen showed a sensitivity of 100%, specificity of 60% and predictability of 70%. Another cut-off value of >80% of total IR revealed a sensitivity of 100%, specificity of 69%, and predictability of 80%. For 12 newly diagnosed glioblastoma patients, median progression-free survival of the HDRA sensitive group was 21 months, while that of the non-sensitive group was 6 months ($p$=0.07). Conclusion : HDRA for malignant glioma was inferred as a feasible method to predict the chemotherapy response. We are encouraged to launch phase 2 clinical trial with chemosensitivity on HDRA.

• Natural Product Sciences
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• 제17권4호
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• pp.315-320
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• 2011

Routinely applicable HPLC assay procedures for the ganglioside content in various deer antler preparations were established through the creation of a UV-absorbing chromophoric substance - trans-${\alpha},{\beta}$-unsaturated-hexadecene-aldehyde - from the sphingosine moiety in ganglioside molecules by two step chemical reactions. In order to guarantee the assay's accuracy and sensitivity, the HPLC-assay procedure adopted internal reference procedures by mixing cis-${\alpha},{\beta}$-unsaturated-hexadecene aldehyde[V] or cis-3-heptadecene- 1,2-diol[IV] to assay samples. The internal reference compound [IV] or [V] was synthesized in our laboratory starting from mannitol-diacetonide through three or four step organic reactions. This new HPLC-assay procedure was successfully applied to deer antler extracts with good dose-dependent calibration curves at the picomole level of gangliosides.

• 대한의생명과학회지
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• 제26권3호
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• pp.149-156
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• 2020

We developed a simple and rapid method for detecting vancomycin resistance genes, such as vanA and vanB, using loop-mediated isothermal amplification (LAMP). To identify not only vancomycin resistance genes, but also the genus Enterococcus, primers were designed for vanA, vanB, and 16S rRNA. Screening for vancomycin susceptibility in Enterococcus was performed using Etest (bioMérieux Inc). The results of the LAMP assay were compared to those of real-time RT-PCR. The optimal conditions for the LAMP assay were 65℃ for 60 min. The detection limits of the LAMP assay for vanA, and vanB were 2 × 10² copies/reaction. Compared to RT-PCR, the sensitivities and specificities of LAMP for 16S rRNA, vanA, and vanB were 100/100%, 100/100%, and 100/100%, respectively. The vanA genotype-vanB phenotype accounted for 57.5% (46/80) of the vancomycin-resistant Enterococci samples collected from 2016 to 2019. In conclusion, the LAMP assay developed in this study showed high sensitivity and specificity for vancomycin-resistant genes. Moreover, due to the simplicity and rapidity of the LAMP assay, its use can be very useful in clinical microbiology laboratories.

• Nuclear Engineering and Technology
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• 제45권7호
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• pp.921-928
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• 2013

A future nuclear energy system is being developed at Korea Atomic Energy Research Institute (KAERI), the system involves a Sodium Fast Reactor (SFR) linked with the pyro-process. The pyro-process produces a source material to fabricate a SFR fuel rod. Therefore, an isotopic fissile content assay is very important for fuel rod safety and SFR economics. A new technology for an analysis of isotopic fissile content has been proposed using a lead slowing down spectrometer (LSDS). The new technology has several features for a fissile analysis from spent fuel: direct isotopic fissile assay, no background interference, and no requirement from burnup history information. Several calculations were done on the designed spectrometer geometry: detection sensitivity, neutron energy spectrum analysis, neutron fission characteristics, self shielding analysis, and neutron production mechanism. The spectrum was well organized even at low neutron energy and the threshold fission chamber was a proper choice to get prompt fast fission neutrons. The characteristic fission signature was obtained in slowing down neutron energy from each fissile isotope. Another application of LSDS is for an optimum design of the spent fuel storage, maximization of the burnup credit and provision of the burnup code correction factor. Additionally, an isotopic fissile content assay will contribute to an increase in transparency and credibility for the utilization of spent fuel nuclear material, as internationally demanded.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2000년도 추계학술발표대회
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• pp.119-127
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• 2000

The development and performance of a competitive indirect immunoassay for the quantitative measurement of polychlorinated biphenyl compounds in insulating oils is presented. Reagent preparation and the assay characterisation, optimisation and validation steps are described. The dynamic range of the assay for Aroclors 1254 and 1260 in methanol was $50-800 {\mu}g\;ml^{-1}$ with $50\%$ signal inhibition values of 217 and $ 212 {\mu}g\;ml^{-1}$ respectively. Impending legislation in the UK is likely to decree that oils containing $ >50 {\mu}g\;ml^{-1}$ PCB be considered contaminated. Assay sensitivity increased with the degree of PCB chlorination. The assay of structurally related compounds of environmental concern yielded cross-reactivity values of under $0.6\%$. The immunoassay proved reliable for the analysis of transformer oils containing $>70{\mu}g\;ml^{-1}$ PCB, but over-estimated PCB levels in oils containing $<20{\mu}g\;ml^{-1}$ of the analyte with the oils requiring pre-treatment using either solid-phase extraction techniques or washing with KOH-ethanol/sulphuric acid to remove matrix interferents. The analytical performance of the assay was compared against a commercially available semi-quantitative immunoassay kit for PCBs in soil and water.

• 한국임상수의학회지
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• 제19권3호
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• pp.295-298
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• 2002

본 연구에서는 개의 Helicobacter 균속 감염을 비침습적 방법으로 진단하기 위해 분변을 시료로한 PCR 검사법을 확립하고 그 효능을 검정하고자 하였다. 분변 PCR 분석은 분변에서 채취된 DNA에서 Helicobacter 균속의 16S RNA의 특이적인 영역에 반응하는 primer를 이용해 수행하였다. 분변 PCR분석법에 의한 결과와 위조직 검사 법 결과를 비교해 본 결과 분변 PCR분석법은 높은 특이도 (100%)와 민감도(96%)를 나타내었다. 도한 확립된 분볍 PCR분석법을 이용해 국내 애완결들의 위내 Helicobacter 균속 감염율을 조사한 결과 감염율이 72.1%로 나타났다. 이상의 결과 본 연구에서 확립한 분변 PCR 분석법은 개의 Helicobacter 균 감염을 진단할 수 있는 새로운 비침습적 진단방법으로 이용될 수 있으리라 사료된다.

• The Plant Pathology Journal
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• 제38권2호
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• pp.159-166
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• 2022

Barley yellow dwarf virus (BYDV) has been a major viral pathogen causing significant losses of cereal crops including oats worldwide. It spreads naturally through aphids, and a rapid, specific, and reliable diagnostic method is imperative for disease monitoring and management. Here, we established a rapid and reliable method for isothermal reverse transcription recombinase polymerase amplification (RT-RPA) combined with a lateral flow strips (LFS) assay for the detection of BYDV-infected oat samples based on the conserved sequences of the BYDV coat protein gene. Specific primers and a probe for RT-RPA reacted and optimally incubated at 42℃ for 10 min, and the end-labeled amplification products were visualized on LFS within 10 min. The RT-RPA-LFS assay showed no cross-reactivity with other major cereal viruses, including barley mild mosaic virus, barley yellow mosaic virus, and rice black streaked dwarf virus, indicating high specificity of the assay. The sensitivity of the RT-RPA-LFS assay was similar to that of reverse transcription polymerase chain reaction, and it was successfully validated to detect BYDV in oat samples from six different regions and in individual aphids. These results confirm the outstanding potential of the RT-RPA-LFS assay for rapid detection of BYDV.

• Tuberculosis and Respiratory Diseases
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• 제84권3호
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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.