• Title/Summary/Keyword: sensitive detection

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Rapid and Sensitive Detection of Hepatitis C Virus in Clinical Blood Samples Using Reverse Transcriptase Polymerase Spiral Reaction

  • Sun, Wenying;Du, Ying;Li, Xingku;Du, Bo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.3
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    • pp.459-468
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    • 2020
  • This study established a new polymerase spiral reaction (PSR) that combines with reverse transcription reactions for HCV detection targeting 5'UTR gene. To avoid cross-contamination of aerosols, an isothermal amplification tube (IAT), as a separate containment control, was used to judge the result. After optimizing the RT-PSR reaction system, its effectiveness and specificity were tested against 15 different virus strains which included 8 that were HCV positive and 7 as non-HCV controls. The results showed that the RT-PSR assay effectively detected all 8 HCV strains, and no false positives were found among the 7 non-HCV strains. The detection limit of our RT-PSR assay is comparable to the real-time RT-PCR, but is more sensitive than the RT-LAMP. The established RT-PSR assay was further evaluated for detection of HCV in clinical blood samples, and the resulting 80.25% detection rate demonstrated better or similar effectiveness compared to the RT-LAMP (79.63%) and real-time RT-PCR (80.25%). Overall, the results showed that the RT-PSR assay offers high specificity and sensitivity for HCV detection with great potential for screening HCV in clinical blood samples.

Development of Nested PCR Primer Set for the Specific and Highly Sensitive Detection of Human Parvovirus B19

  • Cho, Kyu-Bong
    • Biomedical Science Letters
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    • v.24 no.4
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    • pp.390-397
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    • 2018
  • For the specific detection of human Parvovirus B19 (HuPaV-B19), we designed ten specific PCR primers from 3,800~4,500 nucleotides of HuPaV-B19 complete genome (NC_000883.2). Seventeen candidate PCR primer sets for specific detecting HuPaV-B19 were constructed. In specific reaction of HuPaV-B19, seventeen PCR primer sets showed specific band, however five PCR primer sets were selected basis of band intensity, amplicon size and location. In non-specific reaction with seven reference viruses, four PCR primer sets showed non-specific band, however one PCR primer set is not. Detection sensitivity of final selective PCR primer set was $100fg/{\mu}L$ for 112 minute, and PCR amplicon is 539 base pairs (bp). In addition, nested PCR primer set was developed, for detection HuPaV-B19 from a low concentration of contaminated samples. Selection of nested PCR primer set was basis of sensitivity and groundwater sample tests. Detection sensitivity of final selective PCR and nested PCR primer sets for the detection of HuPaV-B19 were $100fg/{\mu}L$ and $100ag/{\mu}L$ basis of HuPaV-B19 plasmid, it was able to rapid and highly sensitive detection of HuPaV-B19 than previous reports. We expect developed PCR primer set in this study will used for detection of HuPaV-B19 in various samples.

Real-time Detection of Magnetic Beads using Highly Sensitive Spin-valve Devices for a Chip-cytometer

  • Roh, Jong-Wook;Son, Oh-Taek;Jung, Hyo-Il;Lee, Woo-Young
    • Proceedings of the Korean Magnestics Society Conference
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    • 2010.06a
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    • pp.189-190
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    • 2010
  • Our results demonstrate the possibility of implementing a chip-cytometer for biological applications using high-sensitive spin-valve devices integrated with a microfluidic channel. Further studies will be extended to the real-time detection of animal cells coated with magnetic beads for the biological applications.

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Chemiluminogenic imaging for highly sensitive detection of DNA

  • Kai, Masaaki;Hirayama, Fumiko;Ohta, Kazuko;Kabashima, Tsutomu;Lee, Myung-Koo
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.407.2-407.2
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    • 2002
  • We have been studing sensitive non-enzymatic chemiluminescence (CL) imaging methods for the detection of DNA. For one of our methods, a unique chemical derivatization reagent. 3'. 4', 5'-trimethoxyphenylglyoxal (TMPG) was utilized. This reagent reacted specifically with guanine bases in nucleic acids to quickly produce a chemiluminescent derivative under mild reaction conditions. (omitted)

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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.

Enzyme Based Biosensors for Detection of Environmental Pollutants-A Review

  • Nigam, Vinod Kumar;Shukla, Pratyoosh
    • Journal of Microbiology and Biotechnology
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    • v.25 no.11
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    • pp.1773-1781
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    • 2015
  • Environmental security is one of the major concerns for the safety of living organisms from a number of harmful pollutants in the atmosphere. Different initiatives, legislative actions, as well as scientific and social concerns have been discussed and adopted to control and regulate the threats of environmental pollution, but it still remains a worldwide challenge. Therefore, there is a need for developing certain sensitive, rapid, and selective techniques that can detect and screen the pollutants for effective bioremediation processes. In this perspective, isolated enzymes or biological systems producing enzymes, as whole cells or in immobilized state, can be used as a source for detection, quantification, and degradation or transformation of pollutants to non-polluting compounds to restore the ecological balance. Biosensors are ideal for the detection and measurement of environmental pollution in a reliable, specific, and sensitive way. In this review, the current status of different types of microbial biosensors and mechanisms of detection of various environmental toxicants are discussed.

Context-sensitive Word Error Detection and Correction for Automatic Scoring System of English Writing (영작문 자동 채점 시스템을 위한 문맥 고려 단어 오류 검사기)

  • Choi, Yong Seok;Lee, Kong Joo
    • KIPS Transactions on Software and Data Engineering
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    • v.4 no.1
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    • pp.45-56
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    • 2015
  • In this paper, we present a method that can detect context-sensitive word errors and generate correction candidates. Spelling error detection is one of the most widespread research topics, however, the approach proposed in this paper is adjusted for an automated English scoring system. A common strategy in context-sensitive word error detection is using a pre-defined confusion set to generate correction candidates. We automatically generate a confusion set in order to consider the characteristics of sentences written by second-language learners. We define a word error that cannot be detected by a conventional grammar checker because of part-of-speech ambiguity, and propose how to detect the error and generate correction candidates for this kind of error. An experiment is performed on the English writings composed by junior-high school students whose mother tongue is Korean. The f1 value of the proposed method is 70.48%, which shows that our method is promising comparing to the current-state-of-the art.

Damage detection of subway tunnel lining through statistical pattern recognition

  • Yu, Hong;Zhu, Hong P.;Weng, Shun;Gao, Fei;Luo, Hui;Ai, De M.
    • Structural Monitoring and Maintenance
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    • v.5 no.2
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    • pp.231-242
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    • 2018
  • Subway tunnel structure has been rapidly developed in many cities for its strong transport capacity. The model-based damage detection of subway tunnel structure is usually difficult due to the complex modeling of soil-structure interaction, the indetermination of boundary and so on. This paper proposes a new data-based method for the damage detection of subway tunnel structure. The root mean square acceleration and cross correlation function are used to derive a statistical pattern recognition algorithm for damage detection. A damage sensitive feature is proposed based on the root mean square deviations of the cross correlation functions. X-bar control charts are utilized to monitor the variation of the damage sensitive features before and after damage. The proposed algorithm is validated by the experiment of a full-scale two-rings subway tunnel lining, and damages are simulated by loosening the connection bolts of the rings. The results verify that root mean square deviation is sensitive to bolt loosening in the tunnel lining and X-bar control charts are feasible to be used in damage detection. The proposed data-based damage detection method is applicable to the online structural health monitoring system of subway tunnel lining.

Development of an Indirect Non-Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Cronobacter muytjensii in Infant Formula Powder (유아용 조제분유 내 Cronobacter muytjensii 검지를 위한 간접 비경합 면역분석법의 개발)

  • Song, Xinjie;Kim, Myunghee
    • The Korean Journal of Food And Nutrition
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    • v.26 no.4
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    • pp.936-944
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    • 2013
  • Cronobacter muytjensii is an important foodborne pathogen as a potential risk in infant formula powder (IFP). To develop a new and sensitive method for the detection of Cronobacter spp. in IFP, an immunoglobulin G (IgG) specific for C. muytjensii (formerly known as Enterobacter sakazakii ATCC 51329) was developed. Further, an indirect noncompetitive enzyme-linked immunosorbent assay (INC-ELISA) was developed by using the anti-C. muytjensii IgG. As a result, this newly developed INC-ELISA method was found very sensitive for C. muytjensii with detection limit of $6.5{\times}10^3CFU/ml$ in pure culture and 1 cell/25 g of IFP. This INC-ELISA method also displayed excellent specificity for C. muytjensii showing no cross-reactivity with other strains of Cronobacter genus and 11 other foodborne pathogenic strains. These results show that the developed INC-ELISA method was very sensitive, efficient, and rapid for the detection of C. muytjensii. Hence, this method could be applied to the development of diagnostic kits for the rapid and easy detection of C. muytjensii.

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.