• Title/Summary/Keyword: Single dilution method

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The Comparison of the Accuracy in Preparing Small Vancomycin Dosage : Single vs. Double Dilution Method (소용량 Vancomycin 주사액 조제방법의 차이에 의한 정확성 비교: 단일희석방법과 이중희석방법)

  • Shin, Hea Young;Suh, Ok Kyung;Lee, Suk Hyang;Shin, Hyun Taek
    • Korean Journal of Clinical Pharmacy
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    • v.7 no.2
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    • pp.86-90
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    • 1997
  • This study was to find a more accurate method fur measuring small vancomycin dosages which are commonly used in neonates by comparing single and double dilution method. For single dilution method, 500 mg of vancomycin powder was accurately measured and reconstituted with 5 ml of distilled water to make a concentration of 100 mg/ml. Volumes of 0.05, 0.1, 0.15, 0.2, 0.3, 0.4, and 0.5 ml, which equal the target dosages of 5, 10, 15, 20, 30, 40, and 50 mg, were measured using syringes made by Shina and each sample was further diluted with 2 ml of $5\%$ dextrose. The solution of 100 mg/ml concentration was further diluted with $5\%$ dextrose to make a concentration of 20 mg/ml. Volumes of 0.25, 0.5, 0.75, 1.0, 1.5, 2.0, and 2.5 ml, which correspond to 5, 10, 15, 20, 30, 40, and 50 mg, were sampled by the same Shina's syringe as in single dilution method and then each sample was further diluted to make a total volume of 10 ml. Each sample was analyzed by HPLC. The measured dosages of each sample in both single and double dilution methods were lower than the target dosages; however, e values in double dilution method were higher than those in single dilution method for seven target dosages. Percent target dosages in single dilution method were 65 to $90\%$, while in double dilution method 91 to $94\%$. Statistically significant difference between two groups was shown in 5, 10, 15, 20, and 40 mg dosages (p<0.05). In conclusion, when preparing small vancomycin dosages lower an 20 mg $(volume{\leq}0.2\;ml)$, using Shina's syringes, the double dilution method has a closer value to the target dosage than single dilution method.

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An Investigation on Dynamic Behaviors of Single Vortex with CO2 Dilution in a CH4-Air Jet Diffusion Flame (CH4공기 제트 확산화염에서 CO2 첨가에 따른 단일 와동의 동적거동에 관한 연구)

  • Hwang, Chul-Hong;Oh, Chang-Bo;Lee, Dae-Yup;Lee, Chang-Eon
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.27 no.9
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    • pp.1209-1219
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    • 2003
  • The dynamic behaviors of the single vortex interacting with $CH_4-Air$ jet diffusion flame are investigated numerically. The numerical method is based on a predict-corrector scheme for a low Mach number flow. A two-step global reaction mechanism is adopted as a combustion model. Studies are conducted in fixed initial velocities for the three cases according as where $CO_2$ is added; (1) without dilution, (2) dilution in fuel stream and (3) dilution in oxidizer stream. A single vortex is generated by an axisymmetric jet, which is made by an impulse of a cold fuel when a flame is developed entirely in a computational domain. The simulation shows that $CO_2$ dilution in fuel stream results in somewhat larger vortex radius, and greater amount of entrainment of surrounding fluid than in other cases. Thus, the dilution of $CO_2$ in fuel stream enhances the mixing in single vortex and increases the stretching of the flame surface. The budgets of the vorticity transport equation are examined to reveal the mechanism of vortex formation when $CO_2$ is added. It is found that, in the case of $CO_2$ dilution in fuel stream, the vortex destruction due to volumetric expansion and the vortex production due to baroclinic torque are more dominant than in other cases.

A Comparative Experiment on the Methods for Antibiotic Sensitivity Test In Vitro (항생제에 대한 세균학적 감수성시험방법에 관한 비교실험(Tetracycline, Neomycin 및 Colistin))

  • Pak, P.W.;Kim, Y.J.
    • The Journal of the Korean Society for Microbiology
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    • v.9 no.1
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    • pp.25-31
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    • 1974
  • A comparative study was performed with 176 cultures of Salmonella organisms on tetracyline, neomycin and colistin in order to find out the relationship between the results obtained from the Ericsson's single disk method and the tube dilution method of antibiotic sensitivity tests which may be carried out in many hospital laboratories. With tetracycline, thirty-three out of 163 cultures of Salmonella typhi were found to be either sensitive or moderate sensitive by means of the disk method and thirty one(ca 94%) out of the thirty three cultures showed less than 1.0 ${\mu}g$ of the Minimal Inhibitory Concentretions(MIC) in the tube-dilution tests, which mean that there were a quite good agreement between the two methods. With neomycin, a hundred and five out of 163 S.typhi were appeared to be either sensitive or moderate sensitive by means of Ericsson's single disk method, among which 103 cultures showed less than 10.0 ${\mu}g$ MIC in the tubedilution method. And also there was a quite correlative pat. terns observed in the result of testing with 13 salmonella cultures other than S. typhi. With colistin, it was hard to observe any particular tendency in the distribution of plotting for 148 cultures showing less the 18 mm in the inhibiting zone diameters between MIC and disk sensitivity patterns except the fifteen, cultures out of 176 salmonella, which appeared to be sensitive in the single disk method and showed less than 1.0 ${\mu}g$ MIC in the tube dilution method.

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Numerical Simulation for Near Field-Behavior of Wastewater Discharged into Stagnant Ambient in Coastal Region (연안지역의 정체수역에서 방류되는 하$\cdot$폐수의 근역거동 수치모의)

  • Kwon, Seok-Jae;Seo, Il-Won
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.17 no.3
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    • pp.166-177
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    • 2005
  • This study developed the jet integral model to analyze the behavior of the wastewater discharge in the near field using the fourth order Runge-Kutta method in order to numerically solve the problems of six ordinary differential equations and six unknowns. This jet integral model used the entrainment hypothesis and the manipulation of sonle shape constant. This study also conducted the hydraulic experiments fnr single horizontal buoyant Jet using LIF through the calibration procedure. The results calculated by the previous models, CORMIX 1 and VISJET, and the proposed jet integral model were compared to the hydraulic experimental results. The centerline trajectories predicted by the proposed model were in good agreements with the experimental results in the transition region whereas the trajectories calculated by the VISJET model agreed well with the measured data in the momentum and buoyancy-dominated regions. The centerline dilution calculated by the proposed model agreed generally with the measured dilution in the intial and transition regions while the centerline dilution predicted by the CORMIX 1 was in good agreements with the experimental results in the momentum and buoyancy-dominated regions.

Standardization of an enzyme-linked immunosorbent assay for detection of antibody to avian reticuloendotheliosis virus (세망내피증 바이러스 항체검출을 위한 ELISA 표준화)

  • Sung, Haan Woo;Lee, Su Jeong
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.569-574
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    • 2005
  • Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

INTERPRETING A SINGLE ANTISTREPTOLYSIN O TEST: A COMPARISON OF THE 'UPPER LIMIT OF NORMAL' AND LIKELIHOOD RATIO METHODS

  • Gray Gregory C.;Struewing Jeffery P.;Hyams Kenneth C.;Escamilla Joel;Tupponce Alan K.;Kaplan Edward L.
    • 대한예방의학회:학술대회논문집
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    • 1994.02b
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    • pp.164-168
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    • 1994
  • Single serologic tests may occasionally influence clinicians in making diagnoses. The antistreptolysin O (ASO) test is a frequently used tool for detecting recent Streptococcus pyogenes infection and is helpful in the diagnosis of diseases like rheumatic fever. Using data from a 1989 prospective study of 600 healthy male military recruits, in which 43% experienced S. pyogenes upper respiratory tract infection (2-dilution rise in ASO), this report compared two methods of interpreting a single ASO titer. Using the 'upper limit of normal' (80 percentile) method, recruits with an ASO titer of greater than 400 showed evidence of recent S. pyogenes infection. This method had a sensitivity and specificity of only 65.9 and 81.9% respectively. In contrast to the 'yes-no'. dichotomy of the 'upper limit of normal' method. the likelihood ratio method statistics were ASO value specific, more consistent with clinical judgment, and better emphasized the caution clinicians must use in interpreting a single ASO test.

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Application of LATE-PCR to Detect Candida and Aspergillus Fungal Pathogens by a DNA Hybridization Assay

  • Gopal, Dhayaalini Bala;Lim, Chua Ang;Khaithir, Tzar Mohd Nizam;Santhanam, Jacinta
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.358-364
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    • 2017
  • Asymmetric PCR preferentially amplifies one DNA strand for use in DNA hybridization studies. Linear-After-The-Exponential-PCR (LATE-PCR) is an advanced asymmetric PCR method which uses innovatively designed primers at different concentrations. This study aimed to optimise LATE-PCR parameters to produce single-stranded DNA of Candida spp. and Aspergillus spp. for detection via probe hybridisation. The internal transcribed spacer (ITS) region was used to design limiting primer and excess primer for LATE-PCR. Primer annealing and melting temperature, difference of melting temperature between limiting and excess primer and concentration of primers were optimized. In order to confirm the presence of single-stranded DNA, the LATE-PCR product was hybridised with digoxigenin labeled complementary oligonucleotide probe specific for each fungal genus and detected using anti-digoxigenin antibody by dot blotting. Important parameters that determine the production of single-stranded DNA in a LATE-PCR reaction are difference of melting temperature between the limiting and excess primer of at least $5^{\circ}C$ and primer concentration ratio of excess primer to limiting primer at 20:1. LATE-PCR products of Candida albicans, Candida parapsilosis, Candida tropicalis and Aspergillus terreus at up to 1:100 dilution and after 1 h hybridization time, successfully hybridised to respective oligonucleotide probes with no cross reactivity observed between each fungal genus probe and non-target products. For Aspergillus fumigatus, LATE-PCR products were detected at 1:10 dilution and after overnight hybridisation. These results indicate high detection sensitivity for single-stranded DNA produced by LATE-PCR. In conclusion, this advancement of PCR may be utilised to detect fungal pathogens which can aid the diagnosis of invasive fungal disease.

DIFFERENCES IN VIBRIO PARAHAEMOLYTICUS DETECTION RATIO BY ANALYTICAL METHODS (측정방법에 따른 장염비브리오균의 검출율의 비교)

  • CHANG Dong-Suck;KIM Young-Man
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.11 no.1
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    • pp.19-24
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    • 1978
  • The differences in Vibrio parahaemolyticus detection ratio were compared between the isolation methods, the Most Probable Number technique and single dilution tube method. During the period from February to October in 1976, 298 samples of sea water, 112 of bottom deposit, 169 of shellfish, and 80 of fish samples collected along the south coastal area of Korea were examined to determine the detection ratio of Vibrio parahaemolyticus. It was often observed that Vibrio parahaemolyticus was detected in higher diluted samples even through negative in lower dilution. Three hundred and forty three samples out of 659 samples submitted to the test by MPN procedure appeared positive for Vibrio parahaemolyticus showing $52\%$ detection ratio. Whereas only 149 samples, corresponding $22.5\%$, were positive for Vibrio parahaemolyticus in the lowest dilution grade. The positive result was $24.5\%$ in the lowest dilution grade and $50\%$ by MPN Procedure in sea water samples, $28.6\%$ and $65.2\%$ in bottom deposit, $22.5\%$ and $56.2\%$ in shellfish and $7.5\%$ and $32.5\%$ fish samples. When tested by triplicate tubes, $61.7\%$ of 149 Vibrio parahaemolyticus positive samples appeared positive in one tube, $28.9\%$ of them were positive in two tubes and $9.4\%$ of them were positive in all three tubes. The detection ratio determined by MPN procedure was more than two times higher than that of single dilution in triplicate tubes.

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Study on Overcoming Interference Factor by Automatic Synthesizer in Endotoxin Test (내독소 검사에서 자동합성장치에 따른 간섭요인 극복에 대한 연구)

  • Kim, Dong Il;Kim, Si Hwal;Chi, Yong Gi;Seok, Jae Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.16 no.2
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    • pp.3-6
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    • 2012
  • Purpose : Samsung medical ceter shall find a cause of the interference factor and suggest a solution for it. Materials and Methods : A sample of $^{18}F$-FDG, radioactive pharmaceuticals produced by TRACERlab MX and FASTlab synthesizer. Gel-clot method uses Positive control tube and single test tube. Kinetic chromogenic method uses ENDOSAFE-PTS produced by Charles River. Results : According to Gel clot method of Endotoxin Tests at FASTlab, both turbidity and viscosity increased at 40-fold dilution and Gel clot was detected. In case of TRACERlab MX, Gel clot was detected in most of samples but intermittently not in a few of them. When using ENDOSAFE-PTS, sample CV (Coefficient of Variation) of FASTlab is 0% at all dilution rates whereas spike CV is 0% at 1-fold dilution, 0~35% at 10-fold, 3.6~12.9% at 20-fold, 5.2~7.1% at 30-fold, 1.1~17.4% at 40-fold, spike recovery; 0% at one-fold, 25 ~ 58% at 10-fold, 50 ~ 86% at 20-fold, 70~92% at 30-fold, and 75~120% at 40-fold. Sample CV of TRACERlab MX, is 0% at all dilution rates whereas spike CV is 1.4~4.8% at one-fold dilution, 0.6~19.9% at 10-fold, spike recovery; 35~72% at one-fold dilution and 77~107% at 10-fold. Conclusion : Gel clot does not seem to occur probably to H3PO4 which engages in bonding with Mg2+ion contributing gelation inside PCT. Dilution which is identical to reducing the amount of H3PO4, could remove interfering effects accordingly. Spike recovery was obtained within 70~150% - recommended values of supplier - at 40-fold dilution even in kinetic chromogenic method.

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Cytochrome b Gene-Based Assay for Monitoring the Resistance of Colletotrichum spp. to Pyraclostrobin

  • Dalha Abdulkadir, Isa;Heung Tae, Kim
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.616-628
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    • 2022
  • Resistance to pyraclostrobin due to a single nucleotide polymorphism at 143rd amino acid position on the cytochrome b gene has been a major source of concern in red pepper field infected by anthracnose in Korea. Therefore, this study investigated the response of 24 isolates of C. acutatum and C. gloeosporioides isolated from anthracnose infected red pepper fruits using agar dilution method and other molecular techniques such as cytochrome b gene sequencing, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and allele-specific polymerase chain reaction (PCR). The result showed that four isolates were resistant to pyraclostrobin on agar dilution method and possessed GCT (alanine) codon at 143rd amino acid position, whereas the sensitive isolates possessed GGT (glycine). Furthermore, this study illustrated the difference in the cytochrome b gene structure of C. acutatum and C. gloeosporioides. The use of cDNA in this study suggested that the primer Cacytb-P2 can amplify the cytochrome b gene of both C. acutatum and C. gloeosporioides despite the presence of various introns in the cytochrome b gene structure of C. gloeosporioides. The use of allele-specific PCR and PCR-RFLP provided clear difference between the resistant and sensitive isolates. The application of molecular technique in the evaluation of the resistance status of anthracnose pathogen in red pepper provided rapid, reliable, and accurate results that can be helpful in the early adoption of fungicide-resistant management strategies for the strobilurins in the field.