Evaluation of the BioFire FilmArray Pneumonia Panel for the Detection of Bacterial Respiratory Pathogens and Antimicrobial Resistance Genes in Endotracheal Aspirate Specimens / 대한임상미생물학회지

Background@#Rapid detection of the causative agents is essential for determining the appropriate treatment for patients with lower respiratory tract infections. We evaluated the performance of the Biofire FilmArray pneumonia panel (FA-PE; BioFire Diagnostics, USA) in the identification of bacterial pathogens and antibiotic resistance genes in endotracheal aspirate specimens. @*Methods@#A total of 43 non-duplicated endotracheal aspirates were included in this study. The performance of the FA-PE was assessed using the routine culture method as the reference standard. @*Results@#The FA-PE demonstrated 92.9% sensitivity and 79.3% specificity for the identification of 15 bacterial targets compared to routine bacterial culture. Four antimicrobial resistance genes in 43 specimens were detected by the FA-PE. The most frequently detected resistance genes were mecA/C and SCCmec in three specimens, followed by CTX-M in one specimen. @*Conclusion@#The FA-PE offers a rapid diagnostic method for lower respiratory tract infections.It may be useful at the early stage of pneumonia, before routine culture and antimicrobial susceptibility results are available.

Development of an Operational Plan for the Liquid Biopsy External Quality Assessment Program in Korea

Background@#Liquid biopsy is a useful assay for the diagnosis, treatment, and prognosis prediction of solid tumors and its clinical application is expanding. Therefore, the need for developing an External Quality Assessment (EQA) protocol for liquid biopsy is increasing. In this study, we developed and implemented the liquid biopsy EQA program for the epidermal growth factor receptor mutation. @*Methods@#We validated the feasibility of the protocol using citrate instead of ethylenediaminetetraacetic acid (EDTA). Additionally, we analyzed the homogeneity and stability of the aliquoted quality control (QC) materials. Mutation-positive QC material with four mutations (exon 19 deletion, L858R, T790M, and exon 20 insertion) was used to make two types of QC materials (low and high) and the wild type material was used for the negative controls. If the EQA results showed consensus in more than 80% of the participating laboratories, the results were reported as acceptable or unacceptable. If not, we reported the results as not graded. @*Results@#Citrate showed equivalent performance to EDTA. Highly mutated QC material and mutation-negative QC material passed the homogeneity and stability test, but low-level mutant specimens showed inconsistent results. In total, 11 laboratories participated, and all of them reported consistent results except for low-grade mutant samples. Thus, the evaluation results were acceptable except for low mutation QC material. @*Conclusions@#The applicability of liquid biopsy is expanding. To obtain accurate test results, EQA is indispensable. Here, QC materials for liquid biopsy EQA were produced, distributed, and had its results analyzed. This study could be the foundation for further development of liquid biopsy EQA.

Effect of the Standardization of Diagnostic Tests on the Prevalence of Diabetes Mellitus and Impaired Fasting Glucose

BACKGROUND: Without standardization of medical laboratory's testing practices, there is an increase in false diagnoses when relying on test results. However, the effect of test standardization is difficult to assess numerically. This study's purpose is to quantify the effect of the standardization level of a laboratory on the prevalence of diabetes mellitus (DM) and impaired fasting glucose (IFG). METHODS: Laboratories were classified into three levels: ‘highly-standardized laboratory,’‘basically-standardized laboratory,’ and ‘non-standardized laboratory.’ Based on the results of Korean External Quality Assessment Scheme (KEQAS), the cutoff values for diagnosis of DM and IFG were recalculated, given false positive and false negative rates. RESULTS: The prevalence of DM and IFG in the population as a whole was estimated using the 2013 Korea National Health and Nutrition Examination Survey (KNHANES) database. When the prevalence of DM from KNHANES was 11.88% (95% confidence interval [CI], 10.59%–13.17%), the proportion with a systematic false error ranged from 10.91% (95% CI, 9.65%–12.17%) to 13.09% (95% CI, 11.74%–14.45%). The prevalence of IFG varied from 13.59% (95% CI, 12.25%–14.91%) to 40.49% (95% CI, 38.54%–42.43%), in contrast to 24.58% (95% CI, 22.85%–26.31%) of the reference value. The prevalence of DM and IFG tended to be over- and under-estimated more as the laboratory standardization level became lower, respectively. CONCLUSION: Our study proved that standardization of clinical laboratory tests is an important factor affecting the prevalence estimation of national disease statistics based on the simulation using KNHANES data.

Comparison of Multilocus Sequence Typing Change Patterns of Vancomycin-Resistant Enterococcus faecium from 2015 to 2017 / 대한임상미생물학회지

BACKGROUND: Multilocus sequence typing (MLST) is useful in determining the long-term evolutionary process and minimizes differences in experimental results across individuals and laboratories. It is also useful in determining evolutionary origins and backgrounds of bacterial species. This study carries out MLST analysis on VanA-type vancomycin-resistant Enterococcus faecium isolated from patient specimens in a single university hospital over nine years in order to observe changes in genetic evolution over time. METHODS: During the years from 2007 to 2015, 44 clinical isolates of vanA-containing E. faecium were collected from Ajou University Hospital in Korea. Species were identified by the VitekII system (bio-Merieux, USA), and antibiotic susceptibility testing was performed by disk diffusion and E-test according to Clinical and Laboratory Standards Institute (CLSI) guidelines. To determine genetic relatedness, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF M/S) was employed. To characterize clonal diversity, MLST analysis was used. RESULTS: All isolates were highly resistant to ampicillin, ciprofloxacin, and vancomycin but showed variable levels of resistance to teicoplanin. The 44 clinical isolates were genetically unrelated according to MALDI-TOF M/S analysis. MLST showed that the clinical isolates harbored 6 sequence types (ST), with ST17 (n=19) being the most common, followed by ST78 (n=13), ST192 (n=6), ST64 (n=4), ST262 (n=1), and ST414 (n=1). CONCLUSION: The MLST analysis showed that the sequence types of most isolates belonged to clonal complex 17 This is consistent with outbreaks in hospitals. We had single observations for ST262 and ST414, suggesting that they were random occurrences. MLST can be useful for speculating the genetic evolution of VanA-containing E. faecium isolates.

Correction: Comparison of Multilocus Sequence Typing Change Patterns of Vancomycin-Resistant Enterococcus faecium from 2015 to 2017 / 대한임상미생물학회지

The period of study in title should have been listed as ‘in the Past Nine Years’. Therefore, we ask to correct ‘from 2015 to 2017’ with ‘from 2007 to 2015’.

Comparison of an international scale method and a log reduction method for monitoring of early molecular response in chronic myeloid leukemia patients

No abstract available.

Epidemiology and Molecular Characterization of Vancomycin-Resistant Enterococcus faecalis / 대한임상미생물학회지

BACKGROUND: Vancomycin-resistant Enterococci (VRE) infections are caused by Enterococcus faecium in about 90% of the cases but can also be caused by Enterococcus faecalis. Thus, this study investigates factors that cause a low isolation rate of vancomycin-resistant E. faecalis (VREfs). To this end, the authors study the clinical traits, resistant gene structure, genomic classification, and molecular characteristics of the virulent factor. METHODS: From January 2001 through September 2011, 17 vanA-containing E. faecalis isolates were collected from hospitalized patients at Ajou University Hospital in Korea. Identification, antimicrobial susceptibility testing, and PCR of van and esp genes were performed. Pulsed-field gel electrophoresis (PFGE) was used for strain typing. PCR and sequencing of the internal regions of Tn1546 were performed for structural analysis of the van gene. RESULTS: Of 4,235 VRE infections, 3,918 (92.5%) were caused by E. faecium, and 95 (2.2%) were caused by E. faecalis. In 67% of VREfs infections, there was a preceding occurrence of E. faecium infection. All isolates were of genotype vanA. Our isolates were divided into three types according to the distribution of IS elements integrated into Tn1546 (types I to IIb). The PFGE results showed no clonal relatedness among isolates. CONCLUSION: Our study found that VREfs infections affect patients who have experienced vancomycin-resistant E. faecium. (VREfm) infection or undergo invasive procedures. The VREfs seems to involve the horizontal transfer of Tn1546 transposon from VREfm.

Increase in the Prevalence of Carbapenem-Resistant Acinetobacter Isolates and Ampicillin-Resistant Non-Typhoidal Salmonella Species in Korea: A KONSAR Study Conducted in 2011

BACKGROUND: Antimicrobial surveillance is important for providing an up-to-date understanding of the epidemiology of antimicrobial resistance and for creating a forum for rational drug development. In this study, we analyzed antimicrobial test data generated in 2011 by hospitals and commercial laboratories participating in the Korean Nationwide Surveillance of Antimicrobial Resistance program (KONSAR). MATERIALS AND METHODS: Data on the results of susceptibility tests conducted in 32 hospitals and two commercial laboratories were analyzed. Data on isolates from patients admitted to an intensive care unit (ICU) and those admitted to other wards were compared. Intermediate susceptibility was not analyzed and duplicate isolates were excluded. RESULTS: Escherichia coli was the most prevalent organism identified in both the hospital and commercial laboratories. Among the hospital isolates, methicillin-resistant Staphylococcus aureus (MRSA), penicillin G-non-susceptible Streptococcus pneumoniae, and ampicillin-resistant Enterococcus faecium remained as prevalent as they were in 2009. The proportion of vancomycin-resistant E. faecium (VR-EFM) slightly decreased from 29% in 2009 to 23% in 2011. Resistance rates of Klebsiella pneumoniae to ceftazidime, cefoxitin, fluoroquinolone, and amikacin were 24%, 14%, 27%, and 8%, respectively. Resistance rates of Pseudomonas aeruginosa to fluoroquinolone, ceftazidime, imipenem, and amikacin were 33%, 20%, 22%, and 16%, respectively, whereas those of Acinetobacter spp. resistance were 71%, 66%, 64, and 51%, respectively. The prevalence of oxyimino-cephalosporin-resistant E. coli and K. pneumoniae, carbapenem-resistant Acinetobacter spp. and P. aeruginosa, MRSA, and VR-EFM among ICU isolates was higher than those among non-ICU isolates. Extended-spectrum beta-lactamase-producing E. coli and K. pneumoniae, imipenem-resistant P. aeruginosa, and VR-EFM were more prevalent among isolates from commercial laboratories than those from hospitals. Resistance rates of K. pneumoniae to ceftazidime and amikacin decreased from 32% and 24% in 2005 to 24% and 8% in 2011, respectively. The resistance rate of P. aeruginosa to amikacin decreased from 22% in 2005 to 16% in 2011. The proportion of imipenem-resistant Acinetobacter spp. increased from 16% in 2005 to 64% in 2011. CONCLUSIONS: The prevalence of MRSA, penicillin G-non-susceptible S. pneumoniae, and ampicillin-resistant E. faecium among clinical isolates tested in laboratories remained high. Multidrug resistance was more prevalent among isolates from ICUs. The prevalence of ceftazidime-resistant and amikacin-resistant K. pneumoniae and amikacin-resistant P. aeruginosa decreased after 2005, while the prevalence of imipenem-resistant Acinetobacter spp. increased.

Effects of Platelet Lysate Preparations on the Proliferation of HaCaT Cells

BACKGROUND: Standard protocols are lacking for the preparation of platelet lysates (PL) as an alternative to using fetal bovine serum as a cell culture supplement. This study aimed to establish optimum conditions for preparing PL for use in cell cultures. METHODS: Cell density in three pooled platelet concentrates (PC) were adjusted to 1x10(12)/L and 2x10(12)/L. PL was prepared from PC by 1 to 3 freeze-thaw (FT) cycles. HaCaT cells were cultured in media supplemented with 5% or 10% PL. Cell numbers were estimated using a Cell Counting Kit-8 (CCK-8; Dojindo Laboratories, Japan). Growth factors were quantified by using the Luminex 200 system (Luminex Corporation, USA). RESULTS: Cell proliferation rates in the presence of PLs were similar when prepared from PCs of both cell densities. The rates were higher in media containing 5% PL than 10% PL when prepared by two FT cycles. Concentrations of vascular endothelial growth factor (VEGF), platelet-derived growth factor-AB/BB (PDGF-AB/BB), PDGF-AA, and epidermal growth factor (EGF) were significantly higher in PL prepared from PC with a cell density of 2x10(12)/L than 1x10(12)/L PC. However, only VEGF and PDGF-AA concentrations in PLs were correlated with HaCaT cell counts. CONCLUSIONS: The 5% PL from PC with a cell density of 1x10(12)/L prepared by two FT cycles treatment was the most effective condition that supported steady HaCaT cell proliferation. Our finding may be useful for preparing PL-supplemented cell culture media.

Distribution of Yeast and Mold Species Isolated from Clinical Specimens at 12 Hospitals in Korea during 2011 / 대한임상미생물학회지

BACKGROUND: The incidence of fungal infections varies among hospitals and between different time periods. We performed a nationwide survey in Korea to investigate the distribution of yeast and mold species recovered from clinical specimens. METHODS: The distributions of clinical isolates of yeast and mold species obtained from 12 university hospitals between January and December 2011 were evaluated relative to the hospital and specimen type. RESULTS: A total of 39,533 fungal isolates (37,847 yeast and 1,686 mold isolates) were obtained. C. albicans was the predominant species (49.4%) among the yeast isolates from all clinical specimens, followed by C. glabrata (7.2%) and C. tropicalis (6.5%). For 5,248 yeast isolates from sterile body fluids, blood was the most common source of yeasts (71.1%), followed by peritoneal fluid (9.4%). Although C. albicans was the predominant species at all but two hospitals, the rate of non-albicans Candida species varied from 71.2% to 40.1%, depending on the hospital. The yeast species recovered most frequently from the sterile body fluids was C. albicans (41.7%), followed by C. parapsilosis (17.8%) and C. glabrata (14.4%), while that from non-sterile sites was C. albicans (50.7%), followed by C. glabrata (6.0%) and C. tropicalis (5.5%). For mold-forming fungi, Aspergillus species (62.3%) were most common, followed by Trichophyton species (15.4%). Respiratory specimens were the most common source of molds (39.6%), followed by abscesses/wounds (28.4%) and tissues (17.5%). CONCLUSION: The rank order of distribution for different fungal species varied among hospitals and specimen types. Continual national surveillance programs are essential for identifying possible changes in fungal infection patterns.

A Case Report of Mycobacterium abscessus Peritonitis in a Patient on Continuous Ambulatory Peritoneal Dialysis / 대한임상미생물학회지

Mycobacterium is an uncommon cause of peritonitis in patients receiving peritoneal dialysis (PD), and the incidence of nontuberculous mycobacterium (NTM) peritonitis is even rarer since the majority of mycobacterial peritonitis cases are caused by Mycobacterium tuberculosis. However, NTM peritonitis has been known to result in a high mortality rate with delayed diagnosis and treatment. In this study, we report a case of Mycobacterium abscessus peritonitis in a 52-year-old male under continuous ambulatory peritoneal dialysis (CAPD).

Methods for Flow Cytometric Analysis of T Cell Subsets in HIV-infected Patients: 2-Color versus 4-Color

BACKGROUND: Blood CD4+ T-lymphocyte (T4) count is a major clinical marker for the diagnosis and management of AIDS, and flow cytometry is considered the gold standard for T4 enumeration. Our aim was to compare the 2-color and 4-color flow cytometric methods for T-cell subset analysis in HIV-infected patients. METHODS: T-cell subsets such as T3, T4, T8, and CD3+CD4-CD8- double negative T cells (DN T) were analyzed from the whole blood of 40 HIV-infected patients by using both 2-color and 4-color methods on a Cytomics FC500 analyzer. Statistical analyses using simple linear regression, paired t-tests, and Bland-Altman plots were performed. RESULTS: The measured T3 (%), T4 (%), T4 (/microL), T8 (%), T8 (/microL), and DN T (%) differed significantly between the 2 methods (P<0.05), whereas the T4/T8 ratio did not. T3 (%), T4 (%), T4 (/microL), T8 (%), T8 (/microL), and T4/T8 measured by the 2 methods showed good correlation, with correlation coefficients above 0.96, whereas DN T (%) did not. The mean differences in T4 (%) and T8 (%) were 0.39% (limit of agreement (LoA), -1.64~2.43) and 1.26% (LoA, -3.37~5.89), respectively. CONCLUSIONS: Although there were statistically significant differences in the T cell subsets measured between the 2 methods, the differences were minor, and the 2 methods showed good correlation. As confirmed in this study, DN T (%) estimated by the 2-color method is lower than the actual value. We suggest that although the 2 methods can be used interchangeably, the 4-color method is recommended for the analysis of some specific subpopulations such as DN T (%).

In Vitro Fluconazole and Voriconazole Susceptibilities of Candida Bloodstream Isolates in Korea: Use of the CLSI and EUCAST Epidemiological Cutoff Values

BACKGROUND: At present, the clinical breakpoints (CBPs) of both fluconazole and voriconazole are available only for 3 common Candida species in the Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) methods. Epidemiological cutoff values (ECVs) were recently applied to both methods to detect the emergence of acquired resistance (i.e., non-wild-type isolates) among 5 common Candida species. METHODS: We performed a nationwide study to determine the fluconazole and voriconazole susceptibility of Candida bloodstream isolates (BSIs) using both the CLSI and EUCAST methods. A total of 423 BSIs of 5 Candida species were collected from 8 hospitals. The azole susceptibilities were assessed on the basis of the species-specific CBPs and ECVs. RESULTS: Of the 341 BSIs of 3 common Candida species (i.e., C. albicans, C. tropicalis, and C. parapsilosis), 0.3% and 0.9%, 0.0% and 1.5% of isolates were categorized as fluconazole and voriconazole resistant according to the CLSI and EUCAST CBPs, respectively. Of 423 total BSIs, 1.4% and 2.6% had fluconazole minimum inhibitory concentrations (MICs) exceeding the ECVs according to the CLSI and EUCAST, respectively; 1.0% and 2.1% had voriconazole MICs exceeding the ECVs according to the CLSI and EUCAST, respectively. Categorical agreement between the methods using ECVs was 98.3% for fluconazole and 98.3% for voriconazole. CONCLUSIONS: The EUCAST and CLSI methods using ECVs provide highly concordant results. Moreover, non-wild-type isolates with possibly acquired azole resistance were rare among the BSIs of 5 common Candida species in Korea.

In Vitro Fluconazole and Voriconazole Susceptibilities of Candida Bloodstream Isolates in Korea: Use of the CLSI and EUCAST Epidemiological Cutoff Values

BACKGROUND: At present, the clinical breakpoints (CBPs) of both fluconazole and voriconazole are available only for 3 common Candida species in the Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST) methods. Epidemiological cutoff values (ECVs) were recently applied to both methods to detect the emergence of acquired resistance (i.e., non-wild-type isolates) among 5 common Candida species. METHODS: We performed a nationwide study to determine the fluconazole and voriconazole susceptibility of Candida bloodstream isolates (BSIs) using both the CLSI and EUCAST methods. A total of 423 BSIs of 5 Candida species were collected from 8 hospitals. The azole susceptibilities were assessed on the basis of the species-specific CBPs and ECVs. RESULTS: Of the 341 BSIs of 3 common Candida species (i.e., C. albicans, C. tropicalis, and C. parapsilosis), 0.3% and 0.9%, 0.0% and 1.5% of isolates were categorized as fluconazole and voriconazole resistant according to the CLSI and EUCAST CBPs, respectively. Of 423 total BSIs, 1.4% and 2.6% had fluconazole minimum inhibitory concentrations (MICs) exceeding the ECVs according to the CLSI and EUCAST, respectively; 1.0% and 2.1% had voriconazole MICs exceeding the ECVs according to the CLSI and EUCAST, respectively. Categorical agreement between the methods using ECVs was 98.3% for fluconazole and 98.3% for voriconazole. CONCLUSIONS: The EUCAST and CLSI methods using ECVs provide highly concordant results. Moreover, non-wild-type isolates with possibly acquired azole resistance were rare among the BSIs of 5 common Candida species in Korea.

Antifungal Susceptibility to Amphotericin B, Fluconazole, Voriconazole, and Flucytosine in Candida Bloodstream Isolates from 15 Tertiary Hospitals in Korea

The in vitro antifungal susceptibility of 636 Candida bloodstream isolates collected from 15 tertiary hospitals in Korea was determined using the Vitek-2 yeast susceptibility system (bioMerieux, France). Overall susceptibility rates were 98.1%, 95.9%, 99.1%, and 97.3% for amphotericin B, fluconazole, voriconazole, and flucytosine, respectively. The results show that the rates of resistance to 4 antifungal drugs remain low among Candida bloodstream isolates in Korea.

Change of Antimicrobial Susceptibility among Escherichia coli Strains Isolated from Female Patients with Community-Onset Acute Pyelonephritis

PURPOSE: There is a concern on which antimicrobials are appropriate as empirical agents for community-onset acute pyelonephritis (APN) in regions where the fluoroquinolone resistance rate is high, such as in Korea. MATERIALS AND METHODS: Three hundred and two strains of E. coli in 2001-2002 and 349 strains in 2008-2009 were isolated from the urine cultures of female adult APN patients, and the antimicrobial susceptibility was compared according to each study period. All the patients were classified as uncomplicated or complicated APN, and a subgroup analysis was done thereafter. RESULTS: The E. coli strains isolated in 2008-2009 showed improved susceptibility to trimethoprim-sulfamethoxazole compared to those isolated in 2001-2002. However, the third generation cephalosporin and gentamicin susceptibility was worsened. Of the 232 isolates from the uncomplicated APN patients, there was no difference between the two different time periods. On the other hand, of the 419 isolates from the complicated APN patients, the susceptibility to third generation cephalosporin, gentamicin and ciprofloxacin was significantly worsened. CONCLUSION: The antimicrobial susceptibility of E. coli changed over the study period, however, this change occurred mainly in the complicated APN patients. In Korea, ciprofloxacin is still useful as an empirical agent for uncomplicated APN patients, but this is not the case for patients with complicated APN because of high resistance rate to ciprofloxacin in these patients. For the complicated APN patients, the rate of resistance to ciprofloxacin is already more than 30%.

Characterization of a Vancomycin-resistant Enterococcus faecium Outbreak Caused by 2 Genetically Different Clones at a Neonatal Intensive Care Unit

In July 2010, we identified an outbreak of vancomycin-resistant enterococci (VRE) in our 26-bed neonatal intensive care unit. We performed an epidemiological investigation after clinical cultures of 2 neonates were positive for VRE. Identification, susceptibility testing, and molecular characterization were performed. Cultures of 3 surveillance stool samples of inpatients and 5 environmental samples were positive for VRE. All isolates were identified as Enterococcus faecium containing the vanA gene. Two distinct clones were identified by performing pulsed-field gel electrophoresis. The 2 clones exhibited different pulsotypes, but they represented identical Tn1546 types. Two sequence types, ST18 and ST192, were identified among all of the isolates with multilocus sequence typing. Our investigation determined that the outbreak in the neonatal intensive care unit was caused by 2 genetically different clones. The outbreak may have occurred through clonal spread and horizontal transfer of the van gene.

A Case of Misidentification of Dermabacter hominis as Listeria grayi / 대한임상미생물학회지

Listeria grayi is a catalase-positive, non-spore forming, and glucose-fermenting Gram-positive rod. L. grayi is widely distributed in environments such as soil, water and fresh food. Human infection by L. grayi is very rare, and there have been no cases reported in Korea, and only two cases worldwide. Dermabacter hominis is a relatively new species belonging to the coryneform bacteria and is a component of the normal human skin flora. D. hominis is a non-motile, glucose-fermenting, Gram-positive rod that has similar biochemical characteristics to L. grayi. The authors of the present study report a case initially misidentified as L. grayi via a traditional morphological and biochemical identification method but that was subsequently confirmed as D. hominis using sequence analysis of 16S rRNA.

Evaluation of a Newly Developed Multiplex Real-time PCR Assay for the Detection of Vancomycin-Resistant Enterococci from Rectal Swabs / 대한임상미생물학회지

BACKGROUND: Asymptomatic vancomycin-resistant enterococci (VRE) colonization precedes infection. VRE-colonized patients serve as silent reservoirs of enterococci that go on to colonize other patients. Rapidly identifying colonized patients is crucial to prevent the spread of VRE. The culture-based method of VRE screening is time-consuming. We evaluated the diagnostic performance of a recently developed multiplex real-time PCR for the detection of VRE. METHODS: We obtained 105 rectal swabs from patients who were being monitored for carriage of VRE. After 24 hour incubation of swabs in enterococcosel broth (EB) supplemented with 6 microg/mL vancomycin, multiplex real-time PCR was performed using the Anyplex(TM) VanR Real-time Detection (VanR) kit (Seegene, Inc., Seoul, Korea). The results of multiplex real-time PCR were compared to those of culture. We evaluated the specificity and detection limits of multiplex real-time PCR using VanR for VRE. RESULTS: A total of 96/105 (91.4%) samples were VRE positive according to multiplex real-time PCR with EB while 85/105 (80.9%) samples were positive in culture. Eleven discordant results (10.4%) (multiplex real-time PCR positive, culture negative) were noted. All non-enterococcal bacteria and vancomycin-susceptible enterococci were negative. The DNA detection limits of VanR were 0.035 pg per reaction (3 microL) for Enterococcus faecium and 0.35 pg for Enterococcus faecalis. CONCLUSION: The application of multiplex real-time PCR after EB incubation allows rapid and sensitive detection in 26-28 hours for VRE screening from rectal swabs. This method could facilitate the timely implementation of contact isolation to prevent the spread of VRE.

The Effect of Freezing-thawing Activated Platelet Rich Plasmas (PRP) on the Proliferations of Bacteria / 대한수혈학회지

BACKGROUND: Fresh platelet rich plasma (PRP) gel has been reported to have anti-bacterial properties. We evaluated the anti-bacterial effects of liquid type activated PRP (tAPRP) using thrombin and heparin treatment after a freezing-thawing (F-T) procedure, using a disk diffusion method. METHODS: PRP and platelet poor plasma (PPP) were prepared from CPDA-1 anticoagulated blood received from 20 donors. PRP was concentrated to 8 times the base platelet counts of donors for the first trial and to 11 times the base platelet counts of donors for the second trial. Both F-T PRP and F-T PPP were divided into a nonactivated group and an activated (tA) group, which was then treated with bovine thrombin and CaCl2, and heparin was added to prevent gel formation. The anti-bacterial effects of F-T PRP, F-T PPP, F-T tAPRP with heparin and F-T tAPPP with heparin on S. aureus and P. aeruginosa were evaluated using a disk-diffusion and direct dropping method. All experiments were duplicated. RESULTS: The inhibited diameters resulting for S. aureus and P. aeruginosa, using the disk-diffusion and direct dropping method, were zero for all 20 sets of results for F-T PRP, F-T PPP, F-T tAPRP with heparin and F-T tAPPP with heparin. CONCLUSION: No anti-bacterial effects were detected for S. aureus or P. aeruginosa in the F-T PRP, F-T PPP, F-T tAPRP with heparin and F-T tAPPP with heparin. This negative result may be due to the F-T treatment and/or because liquid instead of gel form of PRP was used. The use of the disk diffusion method for the determination of anti-bacterial ability of PRP may also be a factor in the negative study results.