• Korean Journal of Clinical Laboratory Science
• /
• v.45 no.1
• /
• pp.16-20
• /
• 2013

Outbreaks of vancomycin-resistant enterococci (VRE) are being reported more frequently in many countries. While seven glycopeptide resistance genotypes have been described in Enterococci, vanA and vanB are the most common resistance genotypes. The aim of this study was to detect antibiotic susceptibilities of 23 Enterococcus faecium strains, which caused an outbreak in a University hospital by a disk diffusion test to investigate the presence of the species specific gene, and the resistant genotypes, vanA and vanB by duplex PCR. PCR for vanA and vanB was performed on 23 enterococci. Twenty three were identified as E. faecium and were tested positive for the vanA genotype. This study will report on the validation of a simple and accurate VRE detection method that can be easily incorporated into the daily routine of a clinical laboratory. Early detection of VRE strains, including those with susceptibility to vancomycin, is of paramount clinical importance as it allows rapid initiation of strict infection control practices, as well as the therapeutic guidance for confirmed infections. The PCR method developed in the present study is simple and reliable for the rapid characterization of VRE.

• Korean Journal of Veterinary Service
• /
• v.22 no.1
• /
• pp.93-101
• /
• 1999

This test process on screening method for the detection of residual antibiotics in milk is simple, economic, sensitive to residual antibiotics and was given approval international organs. Thus, this study was carried out the comparison of Disk assay method and TTC-II method for sensitivity and minimum detectable range of antibiotics in raw milk. The results of this study was summarized as follows ; 1. The number of samples requested for treatment of mastitis was 198 samples. Comparison or analytical results among the methods of TTC-II, disk assay and Delve sp was that TTC-II 37 samples(18.6% ), Disk assay 125samples(63.1%), Delve SP 130 samples(65.7% ) reacted positively. Conformity rate of Delve SP and Disk assay was 70%. 2. Detectable limits of disk assay method in some antibiotics were more sensitive than those of official method(0.05-0.0025ppm in the $\beta$-lactams, 1ppm in two aminoglycoside, 0.2 ppm in one tetracycline, similar in one macrolide) 3. For sensitivity of residual sulfonamides TTC-II was much more sensitive than disk assay. Detectable limits of sulfamethazine and sulfadimethoxine were 30 to 50ppm levels. 4. The best medium preservation period is 1-2 days. 5. Concentration of brome cresol purple related to resistance for B stearothermophilus culture was 24ppm/ml. These results show that disk assay method for screening detection of antibiotics residuces in milk is worthy of use.

• Journal of Microbiology and Biotechnology
• /
• v.33 no.12
• /
• pp.1692-1697
• /
• 2023

Staphylococcus aureus integrated with mecA gene, which codes for penicillin-binding protein 2a, is resistant to all penicillins and other beta-lactam antibiotics, resulting in poor treatment expectations in skin and soft tissue infections. The development of a simple, sensitive and portable biosensor for mecA gene analysis in S. aureus is urgently needed. Herein, we propose a dual-toehold-probe (sensing probe)-mediated exonuclease-III (Exo-III)-assisted signal recycling for portable detection of the mecA gene in S. aureus. When the target mecA gene is present, it hybridizes with the sensing probe, initiating Exo III-assisted dual signal recycles, which in turn release numerous "3" sequences. The released "3" sequences initiate catalytic hairpin amplification, resulting in the fixation of a sucrase-labeled H2 probe on the surface of magnetic beads (MBs). After magnet-based enrichment of an MB-H1-H2-sucrase complex and removal of a liquid supernatant containing free sucrase, the complex is then used to catalyze sucrose to glucose, which can be quantitatively detected by a personal glucose meter. With a limit of detection of 4.36 fM for mecA gene, the developed strategy exhibits high sensitivity. In addition, good selectivity and anti-interference capability were also attained with this method, making it promising for antibiotic tolerance analysis at the point-of-care.

• Korean Journal of Agricultural Science
• /
• v.49 no.4
• /
• pp.795-807
• /
• 2022

The development and commercialization of industrial genetically modified (GM) organisms is actively progressing worldwide, highlighting an increased need for improved safety management protocols. We sought to establish an environmental monitoring method, using real-time polymerase chain reaction (PCR) and propidium monoazide (PMA) treatment to develop a quantitative detection protocol for living GM microorganisms. We developed a duplex TaqMan quantitative PCR (qPCR) assay to simultaneously detect the selectable antibiotic gene, ampicillin (AmpR), and the single-copy Escherichia coli taxon-specific gene, D-1-deoxyxylulose 5-phosphate synthase (dxs), using a direct cell suspension culture. We identified viable engineered E. coli cells by performing qPCR on PMA-treated cells. The theoretical cell density (true copy numbers) calculated from mean quantification cycle (Cq) values of PMA-qPCR showed a bias of 7.71% from the colony-forming unit (CFU), which was within ±25% of the acceptance criteria of the European Network of GMO Laboratories (ENGL). PMA-qPCR to detect AmpR and dxs was highly sensitive and was able to detect target genes from a 10,000-fold (10^-4) diluted cell suspension, with a limit of detection at 95% confidence (LOD_95%) of 134 viable E. coli cells. Compared to DNA-based qPCR methods, the cell suspension direct PMA-qPCR analysis provides reliable results and is a quick and accurate method to monitor living GM E. coli cells that can potentially be released into the environment.

• Journal of Food Hygiene and Safety
• /
• v.35 no.1
• /
• pp.6-12
• /
• 2020

Prevalence of toxin genes and profiles of antibiotic resistance in Vibrio vulnificus were investigated for prevention of Vibrio sepsis and selection of effective antibiotics. A total of 23 V. vulnificus strains were isolated from Vibrio sepsis patients, fish, and water samples collected from fish tanks in restaurants in Jeonnam province during 2015-2017 period. Prevalence of toxin genes including, RtxA, viuB and vvhA were assessed and susceptibilities to 15 different antibiotics were determined. As a result of the toxin gene profile, the RtxA toxin gene was detected in 19 (82.6%) out of 23 strains, and vvhA and viuB toxin genes were positive in all strains. These results showed that V. vulnificus tested in this study possessed at least one more toxin gene, and the toxin gene detection rate was higher than in previous reports. Therefore, there is always a risk of Vibrio sepsis through eating fish or having contact with aquarium water at seafood restaurants. Especially, it was deemed necessary to provide preventive education about Vibrio sepsis for workers in such restaurants. The results of antibiotic susceptibility tests presented 94.4% resistance to cepoxitin antibiotics but all strains showed susceptibility to 14 kinds of antibiotics including chloramphenicol and tetracycline. The currents antibiotic therapy using chloramphenicol and teteracycline against Vibrio sepsis was judged to be useful.

• Pediatric Infection and Vaccine
• /
• v.8 no.2
• /
• pp.199-205
• /
• 2001

Purpose : Since the first report of vancomycin-resistant enterococci(VRE) in 1986, the resistance to vancomycin in enterococci has been increasingly rapidly. In this study, we investigated the clinical manifestations of pediatric patients with VRE and the pattern of the antibiotic use with increasing the rate of VRE in pediatrics Methods : We studied retrospectively 36 pediatric patients who were isolated VRE from January 1998 to December 2000. We classified patients into ICU and non ICU groups and reviewed species of VRE, specimens in which VRE were first detected and procedures performed before VRE detected. Results : We have found that the number of pediatric patients isolated VRE is increasingly annually in this study. In addition, the number of VRE-isolation in the ICU group and in patients who were operated or who underwent active procedures is much higher than that of in the non ICU group and in patients who were taken medication only. Enterococcus faecium is the main species of VRE. VRE showed high resistance to almost all antibiotics except tetracycline, and resistance was closely related to the duration of hospitalization and history of the antibiotic use. The proportion of the cephalosporin use was higher than any other antibiotic before VRE detection. In contrast, that of teicoplanin was higher than any other antibiotic after VRE detection(P<0.05). The cases of superinfection is higher in the ICU group than in non ICU group. Conclusion : In the hospital level, prevention of nosocomial infection through proper administrative policies, through surveillance of high risk VRE regions and prudent antibiotic use can prevent VRE outbreaks and corresponding side effects.

• Pediatric Infection and Vaccine
• /
• v.12 no.2
• /
• pp.124-134
• /
• 2005

Purpose : It has been known that the diagnostic confirmation of group A streptococcal pharyngitis is accompanied with the results of throat culture and/or rapid antigen detection test(RADT). This study was designed to evaluate the usefulness and cost benefits of the RADT in patients with a sore throat compared the empirical antibiotic treated group without using RADT or throat culture with the antibiotic treated group according to the results of RADT test and/or throat culture. Methods : From April 2003 to August 2003, total 369 patients were enrolled this study. They were redistributed into two groups. In one group, the RADT test and throat culture were used and the patients received antibiotic treatment according to the results of test and in the other group, no diagnostic examinations were used and the patients were treated with antibiotics which were chosen empirically. The flow sheet with questionnaire was drawing up and obtained the clinical symptoms, signs and the name of antibiotics that were administered. Results : A total of 244 patients were treated after the throat culture and/or RADT, and 125 patients were treated empirically. The prevalence of bacteriologically confirmed group A streptococcal pharyngitis was 20.1%. The sensitivity and specificity of RADT were 89.8% and 86.1%, respectively. Positive predictive value and negative predictive value were 62.0% and 97.1%, respectively. The rate of antibiotic use was high in both groups. Because the physician used the antibiotics even if the result of RADT was negative. So about 37% of reduction of antibiotics use might be possible if we used antibiotics according to the results of RADT. There were no cost differences between the RADT applied group and the empirically treated antibiotic group if we could reduce the price of RADT to 63% of the current price. Conclusion : The RADT could be applied for the easy and rapid diagnosis and prompt treatment for group A streptococcal pharyngitis, and RADT could reduced the number of antibiotics used if the price of RADT was reduced to 63% of current price. For accurate evaluation of efficacy and cost effect, further controlled study is needed.

• Korean Journal of Poultry Science
• /
• v.40 no.4
• /
• pp.305-313
• /
• 2013

The objective of this study was to investigate occurrence patterns of Clostridium perfringens on different raising periods in broilers. In different raising periods, we investigated the change in the gross lesion and microscopic histological findings of the mucose of the small intestine, colony forming unit (CFU) and the types C. perfringens with PCR assay. According to the gross lesions on the mucose of small intestine with 10-days-old broilers, the non-antibiotic group showed a higher value (0.6) than the antibiotic group (0.0). Whereas 20-days-old broilers with, the antibiotic treatment had a slightly lower value (1.0) than the non-antibiotic group (1.3). In the histological examination on the villi of the small intestine, there was no damage of the villi of the small intestine with 1-day-old broilers in both groups; however, the non-antibiotic group showed a higher value (0.4) than the antibiotic group (0.0) with 10-days-old broilers. In the non-antibiotic group, the CFU of C. perfringens of the fecal samples from the small intestine increased from 10 days of raising broilers and rapidly increase after 20 and 30 days of raising broilers. There was no detection of C. perfringens types with PCR assy in 1-day-old broilers, but we found C. perfringens type A in 10-, 20- and 30-days-old broilers. Although it is possible to raise healthy broilers by using antibiotics, the addition of antibiotics to concentrate feed is prohibited for public health. The results of this study would contribute to proper feeding management through the careful use of antibiotics.

• Korean Journal of Veterinary Service
• /
• v.36 no.4
• /
• pp.311-320
• /
• 2013

The purpose of this study was to evaluate detecting methods and the relationship between tissues and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens. Two groups of broiler chickens were administrated via the drinking water with $50{\mu}g/mL$ and $100{\mu}g/mL$ of enrofloxacin for 5 days, respectively. The concentration of enrofloxacin and metabolic ciprofloxacin in tissues (muscle and kidney) and blood were measured during administration period (for 5 days) and withdrawal period (for 12 days) by high performance liquid chromatography (HPLC) method. Also, all samples were conducted for screening of residues by microbial method using E. coli for quinolone detection and immuno-chromatography method using Smart kit. The relationship between tissues (muscle and kidney) and blood for enrofloxacin and metabolic ciprofloxacin residues in broiler chickens was followed : The levels of enrofloxacin and metabolic ciprofloxacin residues in muscle and kidney were higher 2.9~3.2 folds, 3.6~3.8 folds more than the residues levels in blood, respectively. These results support we can predict the residues in muscle and kidney from the residues in blood. In comparison of detecting methods for antibiotic residues, microbial method using E. coli for quinolone detection and immuno-chromatography method using Smart kit could detect positive reaction at similar or lower concentration than violative concentration of enrofloxacin and metabolic ciprofloxacin in chicken tissues. These results support what two screening methods are useful for screening of quinolone detection in chickens.

• Journal of Korean Society on Water Environment
• /
• v.32 no.6
• /
• pp.670-699
• /
• 2016

In this study, 16 antibiotics were selected from among the top 30 veterinary antibiotics sold in South Korea in 2014, as well as from among the pharmaceuticals targeted by EPA method 1694, in order to review analytical methods for the detection of trace levels of antibiotics in environmental samples: surface water, soils, animal origin foods, and manures. LC-MS/MS was heavily used. In the chromatography for the detection of the selected antibiotics, the $C_{18}$ column was mostly used at the temperature of $30{\sim}40^{\circ}C$. Water and methanol/acetonitrile were commonly chosen as a nonpolar and a polar mobile phase, respectively. Gradient elution was applied to separate multiclass antibiotics. Volatile additives, such as formic acid, acetic acid, and ammonium acetate were mixed with the mobile phase to improve the ionization efficiency of analytes and the sensitivity in MS detection. Electrospray ionization (ESI) was widely used in the LC-MS/MS and positive ionization was preferred to determine the selected antibiotics. A protonated $[M+H]^+$ molecule was selected as a precursor ion, and its two transitions were analyzed, one for quantitative measurement and the other for confirmation. This study reviewed linearity of the calibration curve, recovery, repeatability, method detection limits (MDLs), and method quantification limits (MQLs) for each target compound used to validate the developed analytical methods.