• Title/Summary/Keyword: resistance detection

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Current Methods of Circulating Tumor Cell Detection (순환종양세포 검출 기술)

  • Lim, Minji;Cho, Yoon-Kyoung
    • The Korean journal of helicobacter and upper gastrointestinal research
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    • v.18 no.3
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    • pp.157-161
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    • 2018
  • Liquid biopsy, the analysis of circulating biomarkers from peripheral blood, such as circulating tumor cells (CTCs) and circulating tumor DNA, and exosomes, offers a less invasive, new source of cancer-derived materials that may reflect the status of the disease better and thereby contribute to personalized treatment. Recent advances in microfluidics and molecular analysis technologies have resulted in greatly improved CTC enumeration and detection. In this article, we review commercially available technologies used to isolate CTCs from peripheral blood, including immunoaffinity and label-free, physical property-based isolation methods. Although enormous technological progress has been made, especially within the last decade, only a few CTC detection methods have been approved for routine clinical use. Here, we provide an overview of the current CTC isolation methods and examples of their potential application for early diagnosis, prognosis, treatment monitoring, and prediction of resistance to cancer therapy. Furthermore, the challenges that remain to be addressed before such tools are implemented for routine use in clinical settings are discussed.

Improving the Rapidity and Accuracy of Carbapenem-Resistant Enterobacteriaceae Detection by Shortening the Enrichment Duration

  • Miyoung, Lee
    • Microbiology and Biotechnology Letters
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    • v.50 no.4
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    • pp.584-591
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    • 2022
  • Identifying carbapenem-resistant Enterobacteriaceae (CRE) is necessary to prevent nosocomial CRE infection outbreaks. Here, a rapid identification method with reduced enrichment time was developed without compromising accuracy. A total of 49 rectal swabs requested for CRE screening at the Department of Diagnostic Medicine at Hospital B in Busan, Korea, were included in this study. Specimens were inoculated on MacConkey and CHROMID Carba media either directly or following enrichment for 3, 6, and 24 h in 100 μl trypticase soy broth containing an ertapenem disk. The enriched cultures were further inoculated on CHROMID Carba or MacConkey media containing an ertapenem disk. In total, 19 CRE and 5 carbapenem-intermediate Enterobacteriaceae isolates were obtained from the 49 swabs. Among the 19 CRE isolates, carbapenemase-producing Enterobacteriaceae constituted 13 strains. Moreover, of the 19 CRE isolates, 16 (81.25%) and 17 (88.24%) were identified from the direct cultures on MacConkey and CHROMID Carba media, respectively. After 3 h of enrichment, the proportions of the CRE identified in the media were: MacConkey medium, 16/19 (81.25%); CHROMID Carba medium, 17/19 (88.24%); and MacConkey medium containing an ertapenem disk, 17/19 (88.24%). The detection rates after 6 h of enrichment were the same for all three media (19/19 strains, 100%), whereas those after 24 h of enrichment were 21, 22, and 24 strains, respectively, but included false positives. These findings suggest that a 6-h enrichment before inoculation on the CHROMID Carba medium is optimal for the rapid and accurate detection of CRE in clinical samples.

Development of a Novel Immunochromatographic Assay for Rapid Detection of OXA-23 β-lactamase-producing Acinetobacter baumannii

  • Ji, Gil Young;Song, Hyung Geun;Jo, Mi Young;Hong, Seung Bok;Shin, Kyeong Seob
    • Biomedical Science Letters
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    • v.22 no.2
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    • pp.29-36
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    • 2016
  • Among the several agents causing carbapenem resistance of Acinetobacter baumannii, the most common cause is OXA-23 ${\beta}$-lactamase, which is known to hydrolyze carbapenem. To effectively control dissemination of carbapenem-resistant Acinetobacter baumannii (CRAB), development of both rapid and easy-to-use detection methods are required. The aim of this study is to develop a novel immunochromatographic assay (ICA) for rapid detection of OXA-23 ${\beta}$-lactamase. Of the seven monoclonal antibodies (mAbs) screened by ELISA, four mAbs (4G6, 4H6, 6G4, 9A4) exhibited high reactivity. Of these four specific antibodies, the combination of 6G4/4G6 showed the greatest reactivity and this combination of mAbs (6G4/4G6 mAbs) was used to develop the OXA-23 ${\beta}$-lactamase ICA. Of 102 A. baumannii isolates tested, the OXA-23 ${\beta}$-lactamase ICA results were consistent with PCR analysis except one false positive and one false negative isolate. The overall sensitivity and specificity were 98.36% and 97.56%, respectively. In conclusion, to the best of our knowledge, we have developed the first specific antibody set to detect OXA-23 ${\beta}$-lactamase using an ICA kit. This novel ICA can be used as a reliable and easy-to-use immunological assay for detection of OXA-23 ${\beta}$-lactamase producing CRAB in clinical laboratories.

Evaluation of Reverse Hybridization Assay for Detecting Fluoroquinolone and Kanamycin Resistance in Multidrug-Resistance Mycobacterium tuberculosis Clinical Isolates (다제내성결핵 균주에서 Reverse Hybridization Assay를 이용한 Fluoroquinolone, Kanamycin 신속 내성 검사의 유용성)

  • Park, Chin-Su;Sung, Nack-Moon;Hwang, Soo-Hee;Jeon, Jae-Hyun;Won, Young-Sub;Min, Jin-Hong;Kim, Cheon-Tae;Kang, Hyung-Seok
    • Tuberculosis and Respiratory Diseases
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    • v.72 no.1
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    • pp.44-49
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    • 2012
  • Background: Multidrug-resistant tuberculosis (MDR-TB) is an increasing public health problem and poses a serious threat to global TB control. Fluoroquinolone (FQ) and aminoglycoside (AG) are essential anti-TB drugs for MDR-TB treatment. REBA MTB-FQ$^{(R)}$ and REBA MTB-KM$^{(R)}$ (M&D, Wonju, Korea) were evaluated for rapid detection of FQ and kanamycin (KM) resistance in MDR-TB clinical isolates. Methods: M. tuberculosis (n=67) were isolated and cultured from the sputum samples of MDR-TB patients for extracting DNA of the bacilli. Mutations in genes, gyrA and rrs, that have been known to be associated with resistance to FQ and KM were analyzed using both REBA MTB-FQ$^{(R)}$ and REBA MTB-KM$^{(R)}$, respectively. The isolates were also utilized for a conventional phenotypic drug susceptibility test (DST) as the gold standard of FQ and KM resistance. The molecular and phenotypic DST results were compared. Results: Sensitivity and specificity of REBA MTB-FQ$^{(R)}$ were 77 and 100%, respectively. Positive predictive value and negative predictive value of the assay were 100 and 95%, respectively, for FQ resistance. Sensitivity, specificity, positive predictive value and negative predictive value of REBA MTB-KM$^{(R)}$ for detecting KM resistance were 66%, 94%, 70%, and 95%, respectively. Conclusion: REBA MTB-FQ$^{(R)}$ and REBA MTB-KM$^{(R)}$ evaluated in this study showed excellent specificities as 100 and 94%, respectively. However, sensitivities of the assays were low. It is essential to increase sensitivity of the rapid drug resistance assays for appropriate MDR-TB treatment, suggesting further investigation to detect new or other mutation sites of the associated genes in M. tuberculosis is required.

First Detection of $bla_{IMP-1}$ in Clinical Isolate Multiresistant Acinetobacter baumannii from Korea

  • Jeong Seok-Hoon;Bae Il-Kwon;Sohn Seung-Ghyu;Park Kwang-Ok;An Young-Jun;Sung Kwang-Hoon;Jang Seon-Ju;Heo Myong-Jin;Yang Ki-Suk;Lee Sang-Hee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1377-1383
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    • 2006
  • Among 46 Acinetobacter baumannii isolates collected in 2004, two imipenem-resistant isolates were obtained from clinical specimens taken from patients hospitalized in Busan, Republic of Korea. Two carbapenemase-producing isolates were further investigated to determine the mechanism of resistance. These isolates were analyzed by antibiotic susceptibility testing, microbiological tests of carbapenemase activity, determination of pI, transconjugation test, enterobacterial repetitive consensus (ERIC)-PCR, and DNA sequencing. Two cases of infection by A. baumannii producing the IMP-1 ${\beta}$-lactamase were detected. The isolates were characterized by a modified cloverleaf synergy test and EDTA-disk synergy test. Isoelectric focusing of crude bacterial extracts revealed nitrocefin-positive bands with a pI value of 9.0. PCR amplification and characterization of the amplicons by direct sequencing indicated that the isolates carried a $bla_{IMP-l}$ determinant. The isolates were characterized by a multidrug resistance phenotype, including penicillins, extended-spectrum cephalosporins, carbapenems, and aminoglycosides. These results indicate that the observed imipenem resistance of two Korean A. baumannii isolates was due to the spread of an IMP-1-producing clone. Our microbiological test of carbapenemase activity is simple to screen class B metallo-${\beta}$-lactamase-producing clinical isolates to determine their clinical impact and to prevent further spread. This study shows that the $bla_{IMP-l}$ resistance determinant, which is emerging in Korea, may become an emerging therapeutic problem, since clinicians are advised not to use extended-spectrum cephalosporins, imipenem, and aminoglycosides. This observation emphasizes the importance of having effective control measures in Asian hospitals, such as early detection of colonized patients, isolation procedures, and a judicious use of antibiotics.

Antimicrobial Resistance Patterns and Resistance genes assay of Shigella sonnei Isolated in Korea for Five Years (최근 5년 동안 국내에서 분리된 Shigella sonnei의 항균제 내성 유형과 내성유전자형 분석)

  • Huh, Wan;Lee, Sang-Jo;Kwon, Gi-Seok;Jang, Jong-Ok;Lee, Jung-Bok
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.31-39
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    • 2007
  • This study has been carried out for investigating the relatedness of representative 135 Shigella sonnei strains isolated from 2000 to 2004 by using biotyping and antimicrobial resistance. All strains showed typical biochemical characterisics of Shigella strain. Among 135 strains,79 (58.5%) strains were biotype "g",54 (40.0%) strains were biotype "a" and 2 (1.5%) strains were biotype "e". The results of susceptibility test against 16 antimicrobial agents were like this. Most of strains were susceptible to AN, CIP, C and GM. 129 (95.6%) strains were resistant to SXT, 126 (93.3%) strains were resistant to TE and 122 (90.4%) strains were resistant to SM. One hundred thirty two (97.8%) strains were resistant to more than two antimicrobial agents. R28 type (antimicrobial resistance patterns 28: resistant to AM, SAM, TE, TIC, SXT, K, SM and AmC) were 42 strains (31.1%). The other strains were showed 33 kinds of R patterns. The results of $bla_{TEM}$, sulII, tetA and strA gene detection were coincided with phenotype of antimicrobial resistance by disk diffusion method. But some strains which had sulII and strA genes were not showed the resistance against SXT and SM.

Analysis of Pyrethroid Resistance Allele in Malaria Vector Anopheles sinensis from Malaria High-risk Area (말라리아 위험지역에서 채집된 말라리아 매개모기 Anopheles sinensis의 피레스로이드계 저항성 대립형질 분석)

  • Choi, Kwang Shik;Lee, Seung-Yeol;Hwang, Do-Un;Kim, Heung-Chul;Chang, Kyu-Sik;Jung, Hee-Young
    • The Korean Journal of Pesticide Science
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    • v.20 no.4
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    • pp.286-292
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    • 2016
  • Malaria is mainly transmitted by Anopheles sinensis which is dominant species in malaria high-risk area, northern part of Gyeonggi province in Korea. Pyrethroid insecticide is used for malaria vector, An. sinensis in Korea and the previous investigation consistently reported insecticide resistance from the vector. This study investigated insecticide susceptible and resistant alleles from An. sinensis and the status of malaria vector control in malaria high-risk area. For the study, An. sinensis collected from Paju, Gimpo and Ganghwa were sequenced for kdr detection. In Paju, there was no homozygous susceptibility and all of tested samples had homozygous or heterozygous resistance. There were 6.7% for susceptible homozygosity and 93.3% for resistant homozygosity or heterozygosity in Gimpo. Furthermore, the percentages of homozygous susceptibility and homozygous or heterozygous resistance in Ganghwa were 5.7% and 94.3% respectively. The results showed that the frequency of the insecticide resistance from An. sinensis in malaria high-risk area were increased much more than the previous investigation. Hence, this study suggests that malaria vector control programs should have to be prepared for the management of pyrethroid insecticide resistance.

Detection of MecA Gene in Clinical Isolates of Staphylococcus aureus by Multiplex-PCR, and Antimicrobial Susceptibility of MRSA

  • Lee, Hyean-Woo;Yoon, Joon-Ho;Sohn, Joon-Hyung;Lee, Kyoung-Ho;Yeh, Byung-Il;Park, Deok-Woo;Kim, Hyun-Won;Choi, Jong-Whan
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.354-359
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    • 2003
  • Multiplex-PCR protocols were designed in order to make a rapid identification of MRSA. MecA, femB, and 165 rRNA genes were amplified for making a detection of MRSA. The incidence of MRSA in the clinical isolates of Staphylococcus aureus was examined by using a multiplex-PCR assay. The mecA gene was detected in 266 strains out of 336 clinical isolates of S. aureus, thus the incidence of MRSA was approximately 76.5%. The MRSAs of 247 strains (96.1%) showed resistance to more than eight species of the antimicrobial agents tested. The isolates of MRSA showed 27 different antimicrobial-resistant patterns. The results indicate that many different MRSA strains having high multidrug resistance are actually prevalent in Korea. Also, VISA was screened from the MRSA. Two strains were grown on the BHI agar plate supplemented with $8\;\mu\textrm{g}/ml$ of vancomycin at a frequency of $1/10^8$ colony forming units or higher.

Development of Damage Estimation Method using Sensor of Multiple Function in RC Beam (철근 콘크리트 보에서의 다기능 센서를 이용한 손상 추정법 개발)

  • Kim, Ie-Sung;Park, Kang-Geon;Kim, Wha-Jung
    • Proceeding of KASS Symposium
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    • 2008.05a
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    • pp.184-188
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    • 2008
  • Performance degradation of concrete structures is generally caused by structural deteriorations, such as cracks. It may result in serious defects of concrete structures. Methods of damage detection are used a visual angle of human or non-destructive test, and they are using various sensors. Problems of crack damage detection are occurred to directions of cracks by using 1 axial type of accelerometer in concrete element. In addition, these sensors are not used to occurring fire in RC building. Thermocouple sensors are able to using measurement of temperature in fire, and then deformations of main element and structures are not used. In this study, fundamental studies for development of multiple function sensor using 3 axial type of accelerometer and electric resistance property of thermocouple sensors are discussed estimation to stability of structures when happened form active load or fire, and so on.

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