• Title/Summary/Keyword: Rapid enrichment

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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.

Salmonella sp. 의 신속한 동정을 위한 증진배양의 개선에 관한 연구

  • Kim, Kee-Tae;Kim, Tae-Ue;Yook, Soon-Hak;Pek, Un-Hua
    • Microbiology and Biotechnology Letters
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    • v.24 no.6
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    • pp.647-651
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    • 1996
  • The development of an enrichment method for the rapid and effective identification of Salmonella spp. in sewage or food was studied. As a growth factor for Salmonella, 10 mM cyclic adenosine monophosphate (cAMP) in trypticase soy broth with 0.6% yeast extract (TSBYE) increased cell number five-folds and 0.6% yeast extract in selenite broth increased cell number ten-folds of control. Bile salts in selenite broth was tested for the selection of S. enteritidis in a mixture with Staphylococcus aureus, Pseudomonas aeruginosa, Lactobacillus plantarum and Escherichia coli. The latter four strains were effectively inhibited at 0.1% bile salt. A two-step culture method was used to enrich Salmonella spp.; a primary-enrichment and secondary- enrichment culture. At a primary-enrichment step, selenite broth with 0.6% yeast extract and 10 mM cAMP was used, and at a secondary-enrichment step, 0.1% bile salt was additionally used. Culture times of a primary- enrichment and a secondary-enrichment step were 8 hr and 6 hr, respectively. In this procedure, cell number increased from 10$^{0.3}$ to 10$^{8.5}$ with inhibition of other strains within 14 hr. In the case of an initial cell concentrarion as low as 10$^{-2}$ cfu/ml, a cell number increased to 10$^{7}$ cfu/ml by using a 10 hr primary-enrichment and 6 hr secondary-enrichment procedure.

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Development of a Rapid Enrichment Broth for Vibrio parahaemolyticus Using a Predictive Model of Microbial Growth with Response Surface Analysis (미생물 생장 예측모델과 반응표면분석법을 이용한 Vibrio parahaemolyticus의 신속 증균배지 개발)

  • Yeon-Hee Seo;So-Young Lee;Unji Kim;Se-Wook Oh
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.449-456
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    • 2023
  • In this study, we developed Rapid Enrichment Broth for Vibrio parahaemolyticus (REB-V), a broth capable enriching V. parahaemolyticus from 100 CFU/mL to 106 CFU/mL within 6 hours, which greatly facilitates the rapid detection of V. parahaemolyticus. Using a modified Gompertz model and response surface methodology, we optimized supplement sources to rapidly enrich V. parahaemolyticus. The addition of 0.003 g/10 mL of D-(+)-mannose, 0.002 g/10 mL of L-valine, and 0.002 g/10 mL of magnesium sulfate to 2% (w/v) NaCl BPW was the most effective combination of V. parahaemolyticus enrichment. Optimal V. parahaemolyticus culture conditions using REB-V were at pH 7.84 and 37℃. To confirm REB-V culture efficiency compared to 2% (w/v) NaCl BPW, we assessed the amount of enrichment achieved in 7 hours in each medium and extracted DNA samples from each culture every hour. Real-time PCR was performed using the extracted DNA to verify the applicability of this REB-V culture method to molecular diagnosis. V. parahaemolyticus was enriched to 5.452±0.151 Log CFU/mL in 2% (w/v) NaCl BPW in 7 hours, while in REB-V, it reached 7.831±0.323 Log CFU/mL. This confirmed that REB-V enriched V. parahaemolyticus to more than 106 CFU/mL within 6 hours. The enrichment rate of REB-V was faster than that of 2% (w/v) NaCl BPW, and the amount of enrichment within the same time was greater than that of 2% (w/v) NaCl BPW, indicating that REB-V exhibits excellent enrichment efficiency.

Development of the immunochromatographic strip for the rapid detection of Listeria spp. (Listeria 속균 신속 검출을 위한 면역크로마토그라피 킷트 개발)

  • Jung, Byeong-yeal;Jung, Suk-chan;Kim, Jong-man
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.169-177
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    • 2005
  • We developed the one-step strip based on an immunochromatographic (IC) assay for the rapid detection of Listeria spp. Genus-specific monoclonal antibody to flagella of L. monocytogenes was conjugated with 40 nm colloidal gold particles which were prepared in our laboratory. The specificity of the IC strip was tested with pure cultured bacteria. All strains of the genus of Listeria spp. yielded positive reactions and 12 strains of non-Listeria were negative, resulting in a specificity of 100%. L. monocytogenes was artificially inoculated in raw pork macerated with listeria enrichment broth. And then it was 10-fold diluted from $8.7{\times}10^6$ to 8.7 CFU/ml. L. monocytogenes could be detected at a minimum of $8.7{\times}10^5CFU/ml$ before enrichment, $8.7{\times}10^2CFU/ml$ after primary enrichment and 8.7 CFU/ml after secondary enrichment, respectively. These results indicated that the IC strip exhibited high specificity and sensitivity in the detection of Listeria spp.

The Flame Structure of $CH_4/O_2/N_2$ Premixed Flames on the $O_2$ Enrichment ($CH_4/O_2/N_2$ 예혼합화염에서 산소부화에 따른 화염구조)

  • Lee, Ki-Yong;Nam, Tae-Hyoung;You, Hyun-Seok;Choi, Dong-Soo
    • 한국연소학회:학술대회논문집
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    • 2001.06a
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    • pp.53-59
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    • 2001
  • Experimental measurements are conducted to investigate the structure of flat $CH_4/O_2/N_2$ premixed flames. The flames are simulated using a detailed chemical kinetic mechanism. Four flames established at equivalence ratio = 0.55 are studied with the different $O_2$ enrichment level, ${\Omega}$ = 0.21, 0.25, 0.30, and 0.35. The measured flame speed and species composition profiles are compared with the calculations. Whereas there is overall good agreement between the measurements and predictions, it appears that as the $O_2$ enrichment level is increased the position of the flame is moved toward the exit of the burner and the rapid temperature rise happens near the exit of the burner, and some areas of further refinement in the kinetic mechanism are identified.

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Detection of Methicillin-resistant Staphylococcs aureus from the Anterior Nares of Healthcare Workers in a Intensive Care Unit by Using PBP2a Rapid Kit and Direct Coagulase Test (중환자실에 근무하는 의료인의 전비강에서 PBP2a Rapid Kit와 직접 Coagulase 검사를 이용한 Methicillin-resistant Staphylococcus aureus의 검출)

  • Hong, Seung-Bok;Shin, Kyung-A;Son, Jae-Cheol;Shin, Seob-Kyeong
    • Korean Journal of Clinical Laboratory Science
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    • v.42 no.2
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    • pp.86-91
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    • 2010
  • We evaluated the performance of a novel screening test, PBP2a MRSA rapid kit (Dinona Inc., Iksan, Korea), for methicillin-resistant Staphylococcus aureus (MRSA) based on a immunochromatographic assay. The test is able to detect penicillin-binding protein 2a (PBP2a) using the nasal specimens from health care workers. The nasal specimens were obtained from 69 healthcare workers and were incubated in enrichment broth followed eight hours incubatin in BHI with cefoxitin $4{\mu}g/mL$. These broth were tested by PBP2a Rapid Kit. The enrichment broths were also directly tested for tube coagulase using the conventional identification method. 19 of 22 MRSA showed positive results by PBP2a rapid test and direct coagulase test (the sensitivity for detection of MRSA, 86.36%). While, 8 of 47 non-MRSA showed false positive results for the two tests. All of the 8 non-MRSA which showed false positive were co-colonizing isolates with MRCNS and MSSA. In addition, 46 of 49 methicillin-resistant staphylococci (MRS) showed positive results for PBP2a MRSA rapid kit (the sensitivity for detection of MRS, 93.8%), and all of 20 non-MRS showed negative results (specificity, 100%). The combination of PBP2a MRSA rapid kit and direct coagulase test showed the good sensitivity for detection of MRSA from anterior nares but frequently showed false positive results from the co-colonizing carrier with MRCNS and MSSA.

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THE EVOLUTION OF THE SOLAR NEIGHBORHOOD I: CORRELATIONS OF AGE WITH METALLICITY AND KINEMATIC PARAMETERS

  • Lee, See-Woo
    • Journal of The Korean Astronomical Society
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    • v.15 no.1
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    • pp.19-36
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    • 1982
  • For ${\sim}240$ nearby stars their age and mass were determined and kinematic parameters determined for 362 stars, applying Woolley's three-dimensional potential. Metallicity and kinematic parameters of these stars were correlated with their age, suggesting the slow collapse ($t{\gtrsim}a$ few billion years) of the Galaxy and the initial rapid enrichment in metal abundance (${\Delta}Z{\approx}1/3Z_1$(present) for ${\sim}4{\times}10^8$ yrs). The late slow enrichment rate is given by $d(Z/Z_{\odot})/dt=5.9{\sim}7.0{\pm}3.4$ per Gyr.

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Development of Microfluidic Chip for Enrichment and DNA Extraction of Bacteria Using Concanavalin A Coated Magnetic Particles (Concanavalin A가 코팅 된 자성 입자를 이용한 미생물 농축 및 유전자 추출 칩 개발)

  • Kwon, Kirok;Gwak, Hogyeong;Hyun, Kyung-A;Jung, Hyo-Il
    • Journal of Sensor Science and Technology
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    • v.27 no.4
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    • pp.237-241
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    • 2018
  • The real-time enrichment and detection of pathogens are serious issues and rapidly evolving field of research because of the ability of these pathogens to cause infectious diseases. In general, bacterial detection is accomplished by conventional colony counting or by polymerase chain reaction (PCR) after DNA extraction. As colony counting requires considerable time to cultivate, PCR is an attractive method for rapid detection. A small number of pathogens can cause diseases. Hence, a pretreatment process, such as enrichment is essential for detecting bacteria in an actual environment. Thus, in this study, we developed a microfluidic chip capable of performing rapid enrichment of bacteria and the extraction of their genes. A lectin, i.e., Concanavalin A (ConA), which shows binding affinity to the surface of most bacteria, was coated on the surface of magnetic particles to nonspecifically capture bacteria. It was subsequently concentrated through magnetic forces in a microfluidic channel. To lyse the captured bacteria, magnetic particles were irradiated by a wavelength of 532nm. The photo-thermal effect on the particles was sufficient for extracting DNA, which was consequently utilized for the identification of bacteria. Our device will help monitor the existence of bacteria in various environmental situations such as water, air, and soil.

Rapid diagnosis of experimental listeriosis in mice by polymerase chain reaction (중합효소연쇄반응을 이용한 실험적 리스테리아 감염증의 신속진단)

  • Kang, Ho-jo;Lee, Seong-mi;Suk, Ju-myoung;Lee, Deog-kyu;Son, Won-geun
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.559-564
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    • 1998
  • The polymerase chain reaction(PCR) assay was used for rapid diagnosis from blood and organ samples experimentally infected with Listeria monocytogenes. This method used a pair of primers based on a unique region in the 16S rRNA sequence of L monocytogenes. Procedure A was based on dilution of the blood sample followed by lysis of bacterial cell and direct analysis of the lysate with PCR. In artificially infected blood samples with L monocytogenes, it was possible to detect fewer than 40 cells per ml of blood. However, L monocytogenes was detected low rates on infected organs by the direct PCR. In procedure B, enrichment cultivation was used to increase numbers of bacteria before lysis and PCR. L monocytogenes was detected from 23 samples of 24 liver and spleen, respectively, and 18 samples of 24 blood were found to be positive by PCR on a subset of 72 organ samples, whereas L monocytogenes were detected on 63 organ samples in classical culture technique. It was required to analyze including enrichment steps were 6h and 18h on the procedure A and B, respectively.

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Development of the rapid detection kit for Salmonella spp. using immunochromatographic assay (면역크로마토그라피 기법을 이용한 Salmonella 속균 신속 검출킷트 개발)

  • Jung, Byeong-yeal;Jung, Suk-chan
    • Korean Journal of Veterinary Research
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    • v.45 no.2
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    • pp.191-197
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    • 2005
  • An immunochromatographic (IC) strip for the rapid detection of Salmonella spp. in the enriched sample was developed. Affinity purified Salmonella polyclonal antibody was conjugated with 40 nm colloidal gold particles which were prepared by citrate method in our laboratory. The antigen-antibody-gold complex was captured by Salmonella antibody attached to test line of nitrocellulose membrane during the capillary migration of sample. Specificity of the IC strip was calculated to be 100% (12/12) and sensitivity was 97.6% (41/42) in the test with pure cultured bacteria. Salmonella was artificially inoculated into raw pork macerated with enrichment broth. And then it was 10-fold diluted from $5.2{\times}10^{8}CFU/ml$ to 5.2 CFU/ml. The IC strip could detect $5.2{\times}10^{6}CFU/ml$ before enrichment. However, the lowest limit of detection was 5.2 CFU/ml after overnight incubation. The results indicated that the IC assay was a rapid, economical and simple method with high specificity and sensitivity for the detection of Salmonella spp. without using any equipment.