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2.
Annals of Laboratory Medicine ; : 530-536, 2019.
Article in English | WPRIM | ID: wpr-762442

ABSTRACT

BACKGROUND: Next-generation sequencing is increasingly used for taxonomic identification of pathogenic bacterial isolates. We evaluated the performance of a newly introduced whole genome-based bacterial identification system, TrueBac ID (ChunLab Inc., Seoul, Korea), using clinical isolates that were not identified by three matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems and 16S rRNA gene sequencing. METHODS: Thirty-six bacterial isolates were selected from a university-affiliated hospital and a commercial clinical laboratory. Species was identified by three MALDI-TOF MS systems: Bruker Biotyper MS (Bruker Daltonics, Billerica, MA, USA), VITEK MS (bioMérieux, Marcy l'Étoile, France), and ASTA MicroIDSys (ASTA Inc., Suwon, Korea). Whole genome sequencing was conducted using the Illumina MiSeq system (Illumina, San Diego, CA, USA), and genome-based identification was performed using the TrueBac ID cloud system (www.truebacid.com). RESULTS: TrueBac ID assigned 94% (34/36) of the isolates to known (N=25) or novel (N=4) species, genomospecies (N=3), or species group (N=2). The remaining two were identified at the genus level. CONCLUSIONS: TrueBac ID successfully identified the majority of isolates that MALDI-TOF MS failed to identify. Genome-based identification can be a useful tool in clinical laboratories, with its superior accuracy and database-driven operations.


Subject(s)
Genes, rRNA , Genome , Mass Spectrometry , Seoul
3.
Annals of Laboratory Medicine ; : 119-124, 2018.
Article in English | WPRIM | ID: wpr-713436

ABSTRACT

BACKGROUND: Early detection of tuberculosis (TB) is challenging in resource-poor settings because of limited accessibility to molecular diagnostics. The aim of this study was to evaluate the performance of the loop-mediated isothermal amplification kit (TB-LAMP) for TB diagnosis compared with conventional and molecular tests. METHODS: A total of 290 consecutive sputum samples were collected from May till September, 2015. All samples were processed using the N-Acetyl-L-cysteine (NALC) NaOH method and tested by smear microscopy, solid and liquid culture, real-time PCR, and TB-LAMP. RESULTS: The sensitivity of TB-LAMP for smear-positive and smear-negative samples with culture positivity was 92.0% and 58.8%, respectively. TB-LAMP was positive in 14.9% of TB culture-negative samples; however, all those samples were also positive by real-time PCR. In addition, none of the samples positive for nontuberculous mycobacteria by culture were positive by TB-LAMP. The overall agreement between TB-LAMP and real-time PCR was good; however, the concordance rate was significantly lower for real-time PCR positive samples with Ct values of 30–35. CONCLUSIONS: TB-LAMP could replace smear microscopy and increase TB diagnostic capacity when Xpert MTB/RIF is not feasible because of poor infrastructure.


Subject(s)
Acetylcysteine , Diagnosis , Methods , Microscopy , Nontuberculous Mycobacteria , Pathology, Molecular , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , Sputum , Tuberculosis , Tuberculosis, Pulmonary
4.
Annals of Laboratory Medicine ; : 323-326, 2017.
Article in English | WPRIM | ID: wpr-186609

ABSTRACT

We aimed to analyze the drug resistance patterns of multidrug-resistant and extensively drug-resistant tuberculosis (TB) and the difference of drug resistance among various settings for health care in Korea. The data of drug susceptibility testing in 2009 was analyzed in order to secure sufficient number of patients from various settings in Korea. Patients were categorized by types of institutions into four groups, which comprised new and previously treated patients from public health care centers (PHC), the private sector, and Double-barred Cross clinics (DBC). The resistance rates to first-line drugs were uniformly high in every group. While the resistance rates to second-line drugs were not as high as first-line drugs, there was a pattern that drug resistance rates were lowest for PHC and highest for DBC. The differences of the resistance rates were more prominent for oral second-line drugs. Our findings implied that drug resistance to oral second-line drugs was significantly amplified during multidrug-resistant-TB treatment in Korea. Therefore, an individualized approach is recommended for treating drug-resistant-TB based on susceptibility testing results to prevent acquisition or amplification of drug resistance.


Subject(s)
Humans , Delivery of Health Care , Drug Resistance , Extensively Drug-Resistant Tuberculosis , Korea , Private Sector , Public Health , Tuberculosis, Multidrug-Resistant
5.
Journal of Korean Medical Science ; : 1779-1783, 2017.
Article in English | WPRIM | ID: wpr-225696

ABSTRACT

Close contacts with infectious tuberculosis (TB) are persons at high risk for developing active disease. We preliminarily introduced submillisievert chest computed tomography (CT) scan (effective dose, 0.19–0.25 millisievert) in a contact investigation of multi-drug resistant (MDR)-TB. Baseline CT scan showed minimal nodules or branching opacities in two of six contacts. A two-month follow-up examination revealed a radiologic progression in contact 1, subsequently having the microbiologic diagnosis of MDR-TB at an asymptomatic early stage, whereas nodules transiently increased after 3 months in contact 2, followed by a decrease after one year. Contact 1 was cured after 1.5-year of anti-MDR-TB treatment. In conclusion, early identification of secondary MDR-TB is feasible with submillisievert chest CT scans in contact investigations of MDR-TB, minimizing of MDR-TB transmission and offering a favorable treatment outcome. This was a clinical trial study and was registered at www.ClinicalTrials.gov (Identifier: NCT02454738).


Subject(s)
Humans , Diagnosis , Follow-Up Studies , Thorax , Tomography, X-Ray Computed , Treatment Outcome , Tuberculosis , Tuberculosis, Multidrug-Resistant
6.
Laboratory Medicine Online ; : 189-195, 2017.
Article in Korean | WPRIM | ID: wpr-51170

ABSTRACT

BACKGROUND: Workers in the microbiology laboratories are continuously exposed to the risk of laboratory-associated infections. Tuberculosis (TB) is a frequent laboratory-acquired infection owing to production of cough-generated aerosols with ease and high infectivity of Mycobacterium tuberculosis. This study aims to investigate the current situation of biosafety in Korean TB laboratories. METHODS: We conducted a nationwide survey of laboratories in hospitals conducting TB tests using questionnaires about their facility and management standards. RESULTS: We analyzed data from 52 hospitals nationwide that have a capacity of 100–2,000 beds, of which only two laboratories conduct high risk drug-susceptibility testing on cultured isolates among other test items, whereas six laboratories perform only direct sputum-smear microscopy. The remaining laboratories performed moderate-risk activities/tests, like sample processing for culture. In the majority of these laboratories, there are laboratory medicine specialists who are fully in charge of health checkup programs for laboratorians. The facility and management standards vary widely according to the size of the hospital and risk of TB tests. CONCLUSIONS: Our survey results about the current situation of TB laboratories could be useful as baseline data for preparing biosafety guidelines for all TB laboratories in Korea.


Subject(s)
Aerosols , Korea , Microscopy , Mycobacterium tuberculosis , Specialization , Tuberculosis
7.
Journal of Laboratory Medicine and Quality Assurance ; : 169-193, 2016.
Article in Korean | WPRIM | ID: wpr-65275

ABSTRACT

Annual proficiency surveys were conducted in March, June, and September in 2015 by the Clinical Microbiology Subcommittee of the Korean Association of External Quality Assessment Service. The program covers the sections of bacteriology, advanced bacteriology and mycology, mycobacteriology, and parasitology. Each trial was composed of three sets of different combinations of five bacteria and yeasts. These sets were distributed among laboratories for Gram staining, culture, identification, and antimicrobial susceptibility tests. Five slides with fixed sputum smears were provided as part of each trial for acid-fast bacilli detection. The survey material distribution was section-based. Two survey materials were provided in each trial, while five specimens for mycobacterial culture and identification, five specimens for anti-tuberculosis susceptibility testing and two Mycobacterium tuberculosis strains for rapid detection of rifampin and isoniazid resistance were distributed in the March and June trials. Five virtual microscopy files for stool parasite examination were availed by registered participants in the June trial. Out of the 334 enrolled laboratories, 328 (98.2%), 328 (98.2%), and 329 (98.5%) submitted responses in trials I, II, and III, respectively. Identification of bacteria, namely, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Pseudomonas aeruginosa, Streptococcus pneumoniae, and Vibrio fluvialis by more than 95% of participants was acceptable. Surveillance cultures for vancomycin-resistant enterococci and carbapenem-resistant Enterobacteriaceae were determined accurately by 75.8%–85.3% and 93.1% of the respondents, respectively. Species-level identification of Candida krusei, Candida lusitanae, and Candida guilliermondii was still low at 79.8%, 55.7%, and 42.7%, respectively. Disk diffusion method revealed an unacceptably high false-positive rate of resistance to glycopeptides in E. faecalis and to trimethoprim-sulfamethoxazole in S. pneumoniae. Advanced bacteriology trials revealed unsatisfactory results for species-level identification of moulds. Mycobacterial culture, identification and susceptibility testing, and molecular detection of rifampin and isoniazid resistance were performed exceedingly well by participants. Hymenolepsis diminuta could not be identified by participants, with a correct answer rate of only 46.5% and ‘no parasite seen’ answer rate of only 31.8% for negative specimens. Species-level identification of Candida and moulds was challenging for clinical microbiology laboratories. Disk diffusion method was found to be problematic in testing the susceptibility of microorganisms to glycopeptides and trimethoprim-sulfamethoxazole. Improvement is required in result interpretation of negative specimens in parasitology.


Subject(s)
Bacteria , Bacteriology , Candida , Diffusion , Enterobacteriaceae , Enterococcus faecalis , Escherichia coli , Glycopeptides , Isoniazid , Klebsiella pneumoniae , Korea , Methods , Microscopy , Mycobacterium , Mycobacterium tuberculosis , Mycology , Parasites , Parasitology , Pneumonia , Pseudomonas aeruginosa , Quality Control , Rifampin , Sputum , Streptococcus pneumoniae , Surveys and Questionnaires , Trimethoprim, Sulfamethoxazole Drug Combination , Vancomycin-Resistant Enterococci , Vibrio , Yeasts
8.
Annals of Laboratory Medicine ; : 487-493, 2015.
Article in English | WPRIM | ID: wpr-110968

ABSTRACT

BACKGROUND: Resistance of Mycobacterium tuberculosis to anti-tuberculosis (TB) drugs is almost exclusively due to spontaneous chromosomal mutations in target genes. Rapid detection of drug resistance to both first- and second-line anti-TB drugs has become a key component of TB control programs. Technologies that allow rapid, cost-effective, and high-throughput detection of specific nucleic acid sequences are needed. This study was to develop a high-throughput assay based on allele-specific primer extension (ASPE) and MagPlex-TAG microspheres to detect anti-TB drug resistance mutations. METHODS: DNA samples from 357 M. tuberculosis clinical isolates and H37Rv were amplified by multiplex PCR using four primer sets, followed by multiplex ASPE using 23 TAG-ASPE primers. The products were sorted on the TAG-ASPE array and detected by using the Luminex xMAP system. Genotypes were also determined by sequencing. RESULTS: Genetic drug susceptibility typing by the TAG-ASPE method was 100% concordant with those obtained by sequencing. Compared with phenotypic drug susceptibility testing (DST) as a reference method, the sensitivity and specificity of the TAG-ASPE method were 83% (95% confidence interval [CI], 79-88%) and 97% (95% CI, 90-100%) for isoniazid. For rifampin testing, the sensitivity and specificity were 90% (95% CI, 86-93%) and 100% (95% CI, 99-100%). Also, the sensitivity and specificity were 58% (95% CI, 51-65%) and 86% (95% CI, 79-93%) for ethambutol. CONCLUSIONS: This study demonstrated the TAG-ASPE method is suitable for highly reproducible, cost-effective, and high-throughput clinical genotyping applications.


Subject(s)
DNA , Drug Resistance , Ethambutol , Genotype , Isoniazid , Microspheres , Multiplex Polymerase Chain Reaction , Mycobacterium tuberculosis , Rifampin , Tuberculosis , Viperidae
9.
Annals of Laboratory Medicine ; : 494-499, 2015.
Article in English | WPRIM | ID: wpr-110967

ABSTRACT

BACKGROUND: Tuberculosis (TB) is one of the most serious health problems in Myanmar. Because TB drug resistance is associated with genetic mutation(s) relevant to responses to each drug, genotypic methods for detecting these mutations have been proposed to overcome the limitations of classic phenotypic drug susceptibility testing (DST). We explored the current estimates of drug-resistant TB and evaluated the usefulness of genotypic DST in Myanmar. METHODS: We determined the drug susceptibility of Mycobacterium tuberculosis isolated from sputum smear-positive patients with newly diagnosed pulmonary TB at two main TB centers in Myanmar during 2013 by using conventional phenotypic DST and the GenoType MTBDRplus assay (Hain Lifescience, Germany). Discrepant results were confirmed by sequencing the genes relevant to each type of resistance (rpoB for rifampicin; katG and inhA for isoniazid). RESULTS: Of 191 isolates, phenotypic DST showed that 27.7% (n=53) were resistant to at least one first-line drug and 20.9% (n=40) were resistant to two or more, including 18.3% (n=35) multidrug-resistant TB (MDR-TB) strains. Monoresistant strains accounted for 6.8% (n=13) of the samples. Genotypic assay of 189 isolates showed 17.5% (n=33) MDR-TB and 5.3% (n=10) isoniazid-monoresistant strains. Genotypic susceptibility results were 99.5% (n=188) concordant and agreed almost perfectly with phenotypic DST (kappa=0.99; 95% confidence interval 0.96-1.01). CONCLUSIONS: The results highlight the burden of TB drug resistance and prove the usefulness of the genotypic DST in Myanmar.


Subject(s)
Humans , Drug Resistance , Genotype , Myanmar , Mycobacterium tuberculosis , Rifampin , Sputum , Tuberculosis
10.
Annals of Clinical Microbiology ; : 69-75, 2015.
Article in Korean | WPRIM | ID: wpr-110952

ABSTRACT

BACKGROUND: There have been steady changes and improvements in diagnostic tests for Mycobacterium tuberculosis, so it is necessary to carry out periodic surveys to understand the current situation. The aims of this study were to investigate the changes in principal practices and quality control for M. tuberculosis using a nationwide survey in the Republic of Korea. METHODS: We constructed a questionnaire composed of four subseries with 42 items. We e-mailed this survey to members of the Korean Society of Clinical Microbiology from April to September 2014 and analyzed the replies. RESULTS: Employees at a total of 65 hospital laboratories and 5 commercial laboratories participated in the survey. AFB staining was reportedly performed in all 70 institutions, and fluorescent staining was used as the primary detection method in 59 (84.3%) laboratories. Solid and liquid culture methods for Mycobacterium were performed at 62 (88.6%) and 59 (84.3%) laboratories, respectively. There were 57 laboratories (90.5%) that identified strains growing on primary culture media using a rapid antigen kit or molecular method. The mean values of positive and contamination rates for solid culture media were 8.2% (range 3.7-19.9%) and 4.0% (0.4-8.4%), respectively. In liquid culture, the mean values of positive and contamination rates were 11.5% (4.8-22.3%) and 6.8% (0.3-18.7%), respectively. CONCLUSION: There have been significant changes and improvements in overall mycobacterial testing, especially in the numbers of laboratories using fluorescent staining, liquid culture, and identification of M. tuberculosis cultured media compared with previous surveys in Korea.


Subject(s)
Culture Media , Diagnostic Tests, Routine , Electronic Mail , Korea , Laboratories, Hospital , Mycobacterium , Mycobacterium tuberculosis , Quality Control , Republic of Korea , Tuberculosis
12.
Journal of Laboratory Medicine and Quality Assurance ; : 153-178, 2015.
Article in Korean | WPRIM | ID: wpr-114120

ABSTRACT

Annual proficiency surveys were performed in March, June and September 2014 by clinical microbiology division of The Korean Association of Quality Assurance for Clinical Laboratory. Parasitology part has been newly incorporated in this survey. For each trial, three sets which were composed of different combinations of five bacteria and yeast were distributed for gram stain, culture, identification, and antimicrobial susceptibility tests of general bacteriology and five fixed sputum smear on slides were distributed for acid fast bacilli stain. Two advanced bacteriology survey materials for culture and identification of anaerobic bacteria and mold were distributed to the voluntary participants in every trial and five mycobacterial culture and identification specimens, five anti-tuberculosis susceptibility testing specimens, and two Mycobacterium tuberculosis strains for rapid detection of rifampin and isoniazid resistance were distributed to the voluntary participants in March and June trials. Five virtual microscopic slides for stool parasite examination were open for the registered participants in June trial. A total of 340 laboratories were enrolled and 330 (97.0%), 331 (97.4%), and 331 (97.4%) returned the results on trial I, II, and III, respectively. For bacterial identification, the percent acceptable identification of Burkholderia cepacia, Klebsiella pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pneumoniae, Streptococcus agalactiae, Plesiomonas shigelloides, and Enterococcus faecalis were greater than 95%. Group C and group D Salmonella species challenged as the different sets of M1422 resulted in the acceptable rate lower than 95% because nine participants reported the identification of different sets. Surveillance cultures for methicillin-resistant S. aureus and vancomycin-resistant enterococci were correctly determined by 89.6% and 69.0% of the respondents, respectively. Correct identification to species level of Candida albicans, Candida auris, Candida glabrata, and Candida parapsilosis were 86.1%, 1.6%, 48.1%, and 83.8%. Vancomycin disk diffusion test in S. aureus, missing oxacillin screen or penicillin susceptibility test in S. pneumoniae and lack of reliable methods of quinolone resistance detection in Salmonella species caused unacceptable results in antimicrobial susceptibility testing. Advanced bacteriology trials revealed low performance in species identification of mold. Mycobacterial culture, identification and susceptibility test performance was kept in excellence. The performance of identification of stool parasites was acceptable >90% for detection of helminth eggs and amebic cysts but 28.6% false positive responses resulted from negative specimens. In conclusion, species-level identification of fungi of both candida species and mold were challenging to clinical microbiology laboratories. Vancomycin disk diffusion method for S. aureus and lack of proper penicillin susceptibility test for S. pneumoniae were still common cause of inaccurate results. Virtual microscopic survey has been successfully introduced in parasitology.


Subject(s)
Bacteria , Bacteria, Anaerobic , Bacteriology , Burkholderia cepacia , Candida , Candida albicans , Candida glabrata , Surveys and Questionnaires , Diffusion , Eggs , Enterococcus faecalis , Fungi , Helminths , Isoniazid , Klebsiella pneumoniae , Korea , Methicillin Resistance , Mycobacterium tuberculosis , Ovum , Oxacillin , Parasites , Parasitology , Penicillins , Plesiomonas , Pneumonia , Pseudomonas aeruginosa , Rifampin , Salmonella , Sputum , Staphylococcus aureus , Streptococcus agalactiae , Streptococcus pneumoniae , Streptococcus pyogenes , Vancomycin , Yeasts
13.
Yonsei Medical Journal ; : 572-577, 2015.
Article in English | WPRIM | ID: wpr-38892

ABSTRACT

The trends and types of carbapenemase-producing Gram-negative bacilli were analyzed from clinical specimens collected between 2005 and 2012 at a Korean teaching hospital. The proportions of carbapenem-resistant Acinetobacter spp. increased markedly to 66%. Metallo-beta-lactamase producers significantly decreased and the majority shifted from the bla(VIM-2) type to the bla(IMP-1) type.


Subject(s)
Humans , Acinetobacter/classification , Acinetobacter Infections/drug therapy , Anti-Bacterial Agents/pharmacology , Bacterial Proteins , Carbapenems/pharmacology , Drug Resistance, Microbial , Gram-Negative Bacteria/drug effects , Incidence , Microbial Sensitivity Tests/trends , Population Surveillance , Pseudomonas/classification , Republic of Korea/epidemiology , beta-Lactamases/biosynthesis
14.
Annals of Clinical Microbiology ; : 23-27, 2014.
Article in English | WPRIM | ID: wpr-121391

ABSTRACT

Rapidly growing mycobacteria are ubiquitous in the environment and are increasingly being recognized as opportunistic pathogens. Recently, a new species, Mycobacteium conceptionense, has been validated from the Mycobacterium fortuitum third biovariant complex by molecular analysis. However, there are few reports, and postsurgical wound infection by this species is rare. We report a case of postsurgical wound infection caused by M. conceptionense in an immunocompetent patient that was identified by a sequencing analysis of 16S rRNA, hps65, and rpoB genes.


Subject(s)
Humans , Mycobacterium fortuitum , Mycobacterium , Wound Infection , Wounds and Injuries
15.
Journal of Laboratory Medicine and Quality Assurance ; : 122-139, 2014.
Article in Korean | WPRIM | ID: wpr-131177

ABSTRACT

Annual external quality assessment was performed three times for clinical microbiology division of The Korean Association of Quality Assurance for Clinical Laboratory. For each trial, three sets composed of different combinations of four bacteria and one yeast were distributed for culture, identification, and antimicrobial susceptibility tests. A total of 340 laboratories were enrolled and 330 (97.0%), 331(97.4%), and 331(97.4%) returned the results on trial I, II, and III, respectively. For bacterial identification, the correct identification of gram-negative bacilli, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus capitis, Streptococcus agalactiae, Listeria monocytogenes, and Candida species was greater than 95%. However, correct identification of Staphylococcus lugdunensis, Corynebacterium striatum, Vibrio vulnificus, Aeromonas hydrophila, Cryptococcus neoformans, and Malassezia pachydermatis was relatively less accurate, with values of 95.4%, 89.9%, 50.7%, 91.3%, 93.6%, and 93.9%, respectively. Surveillance cultures for vancomycin-resistant enterococci and methicillin-resistant S. aureus were correctly determined by 95.4% and 93.9% of the respondents, respectively. False carbapenem-resistance due to AmpC beta-lactamase, disk diffusion testing for vancomycin in Staphylococcus species, oxacillin and penicillin susceptibility testing in S. lugdunensis and false imipenem-resistance in Proteus species were common sources of inaccurate results. The accuracy of species identification for Corynebacterium species and Vibrio species requires improvement. Consistent problems occurred with antimicrobial susceptibility testing of vancomycin for Staphylococcus species using the disk diffusion method.


Subject(s)
Aeromonas hydrophila , Bacteria , beta-Lactamases , Candida , Corynebacterium , Cryptococcus neoformans , Surveys and Questionnaires , Diffusion , Korea , Listeria monocytogenes , Malassezia , Methicillin Resistance , Oxacillin , Penicillins , Proteus , Staphylococcus , Staphylococcus aureus , Staphylococcus epidermidis , Staphylococcus lugdunensis , Streptococcus agalactiae , Vancomycin , Vibrio , Vibrio vulnificus , Yeasts
16.
Journal of Laboratory Medicine and Quality Assurance ; : 122-139, 2014.
Article in Korean | WPRIM | ID: wpr-131176

ABSTRACT

Annual external quality assessment was performed three times for clinical microbiology division of The Korean Association of Quality Assurance for Clinical Laboratory. For each trial, three sets composed of different combinations of four bacteria and one yeast were distributed for culture, identification, and antimicrobial susceptibility tests. A total of 340 laboratories were enrolled and 330 (97.0%), 331(97.4%), and 331(97.4%) returned the results on trial I, II, and III, respectively. For bacterial identification, the correct identification of gram-negative bacilli, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus capitis, Streptococcus agalactiae, Listeria monocytogenes, and Candida species was greater than 95%. However, correct identification of Staphylococcus lugdunensis, Corynebacterium striatum, Vibrio vulnificus, Aeromonas hydrophila, Cryptococcus neoformans, and Malassezia pachydermatis was relatively less accurate, with values of 95.4%, 89.9%, 50.7%, 91.3%, 93.6%, and 93.9%, respectively. Surveillance cultures for vancomycin-resistant enterococci and methicillin-resistant S. aureus were correctly determined by 95.4% and 93.9% of the respondents, respectively. False carbapenem-resistance due to AmpC beta-lactamase, disk diffusion testing for vancomycin in Staphylococcus species, oxacillin and penicillin susceptibility testing in S. lugdunensis and false imipenem-resistance in Proteus species were common sources of inaccurate results. The accuracy of species identification for Corynebacterium species and Vibrio species requires improvement. Consistent problems occurred with antimicrobial susceptibility testing of vancomycin for Staphylococcus species using the disk diffusion method.


Subject(s)
Aeromonas hydrophila , Bacteria , beta-Lactamases , Candida , Corynebacterium , Cryptococcus neoformans , Surveys and Questionnaires , Diffusion , Korea , Listeria monocytogenes , Malassezia , Methicillin Resistance , Oxacillin , Penicillins , Proteus , Staphylococcus , Staphylococcus aureus , Staphylococcus epidermidis , Staphylococcus lugdunensis , Streptococcus agalactiae , Vancomycin , Vibrio , Vibrio vulnificus , Yeasts
17.
Annals of Laboratory Medicine ; : 26-30, 2014.
Article in English | WPRIM | ID: wpr-193132

ABSTRACT

BACKGROUND: Early laboratory detection of Mycobacterium tuberculosis is crucial for controlling tuberculosis. We developed a hydrogel mycobacterial culture method that retains the advantages of both solid and liquid methods in terms of speed, cost, and efficiency. METHODS: Mycobacterium bovis bacillus Calmette-Guerin (BCG) suspensions and 200 acid-fast bacilli (AFB)-positive clinical specimens were inoculated in Middlebrook 7H9 liquid media (Becton-Dickinson and Company, USA) and mixed with 75 microL of 9-fluorenylmethoxycarbonyl (Fmoc)-Phe-Phe-OH hydrogel stock solution in an Eppendorf tube just before culture incubation. The mixtures were cultured at 37degrees C for as long as 14 days to monitor culture status. RESULTS: The number of M. bovis BCG increased with time. For 200 AFB smear-positive specimens, 155 of 158 conventional culture-positive specimens and 4 culture-negative or contaminated specimens yielded positive cultures within 14 days. For 128 specimens positive with the liquid culture method, the time to positive culture using the hydrogel method (mean, 12.6 days; range, 7 to 14 days) was significantly shorter than that for conventional liquid culture (mean, 16.2 days; range, 6 to 31 days; P<0.0001). CONCLUSIONS: The hydrogel scaffold culture system is useful for timely, economical, and efficient detection of mycobacteria in clinical specimens.


Subject(s)
Humans , Bacteriological Techniques/methods , Culture Media/chemistry , Early Diagnosis , Hydrogels/chemistry , Mycobacterium tuberculosis/isolation & purification , Tuberculosis/diagnosis
18.
Annals of Laboratory Medicine ; : 31-37, 2014.
Article in English | WPRIM | ID: wpr-193131

ABSTRACT

BACKGROUND: Bacteria of the Mycobacterium abscessus group are the second most common pathogens responsible for lung disease caused by nontuberculous mycobacteria in Korea. There is still a lack of studies investigating the genetic mechanisms involved in M. abscessus resistance to antibiotics other than clarithromycin. This study investigated the characteristics of drug resistance exhibited by M. abscessus clinical isolates from Korea. METHODS: We performed drug susceptibility testing for a total of 404 M. abscessus clinical strains. Subspecies were differentiated by molecular biological methods and examined for mutations in drug resistance-related genes. RESULTS: Of the 404 strains examined, 202 (50.00%), 199 (49.26%), and 3 (0.74%) strains were identified as M. abscessus, M. massiliense, and M. bolletii, respectively. Of the 152 clarithromycin-resistant strains, 6 possessed rrl mutations, while 4 of the 30 amikacin-resistant strains contained rrs mutations, and 5 of the 114 quinolone-resistant strains had gyr mutations. All mutant strains had high minimal inhibitory concentration values for the antibiotics. CONCLUSIONS: Our results showed the distribution of the strains with mutations in drug resistance-related genes was low in the M. abscessus group. Furthermore, we performed drug susceptibility testing and sequence analyses to determine the characteristics of these genes in the M. abscessus group.


Subject(s)
Humans , Anti-Bacterial Agents/pharmacology , Bacterial Proteins/genetics , Clarithromycin/pharmacology , DNA Gyrase/genetics , Drug Resistance, Bacterial , Methyltransferases/genetics , Microbial Sensitivity Tests , Mutation , Mycobacterium/drug effects , Mycobacterium Infections, Nontuberculous/diagnosis , Republic of Korea , Sequence Analysis, DNA
19.
Annals of Clinical Microbiology ; : 80-85, 2014.
Article in Korean | WPRIM | ID: wpr-216375

ABSTRACT

BACKGROUND: Fluoroquinolones (FQs) are important drugs for treating multidrug-resistant tuberculosis (MDR-TB). However, due to widespread use of FQs, the resistance rates to FQs have been increasing among Mycobacterium tuberculosis. Rapid and reliable FQ drug susceptibility testing (DST) is crucial for successful treatment of MDR-TB. In this study, the feasibility of molecular detection of FQ resistance was evaluated. METHODS: A total of 95 MDR-TB isolates were collected from Jan through Oct 2009 at the Korean Institute of Tuberculosis. DST for ofloxacin (OFL), levofloxacin, and moxifloxacin was performed using the Lowenstein-Jensen media absolute concentration method. Minimum inhibitory concentrations (MIC) of these were determined using the broth microdilution method. DNA was extracted from cultured isolates using bead beating method. The quinolone resistance-determining region (QRDR) of gyrA and gyrB were amplified and those sequences were analyzed. RESULTS: Of 95 isolates, 79 were resistant to at least one of FQs. Of these, 71 (89.9%) harbored mutation in the QRDR of gyrA or gyrB. None of FQ susceptible strains possessed any mutation in gyrA or gyrB. Mutations in codon 94 of gyrA were most common; only two isolates had mutation in only the gyrB gene. OFL MICs for isolates with gyrA mutation ranged from 1 to 32 microg/mL, but FQ susceptible isolates showed MICs ranging from < or =0.06 to 0.5 microg/mL. CONCLUSION: Mutation analysis of QRDR of gyrA and gyrB showed 89.9% sensitivity and 100% specificity for detecting FQ resistance in MDR-TB. Therefore, molecular DST can be useful for rapid detection of FQ resistance in MDR-TB.


Subject(s)
Codon , DNA , Fluoroquinolones , Levofloxacin , Microbial Sensitivity Tests , Mycobacterium tuberculosis , Ofloxacin , Tuberculosis , Tuberculosis, Multidrug-Resistant
20.
Annals of Laboratory Medicine ; : 243-246, 2014.
Article in English | WPRIM | ID: wpr-163726

ABSTRACT

Mycobacterium neoaurum is rapidly growing mycobacteria that can cause human infections. It commonly causes bloodstream infections in immunocompromised hosts, and unlike other mycobacteria species, it rarely causes pulmonary infections. We confirmed the first pulmonary infection case in Korea caused by M. neoaurum using full-length 16S rRNA gene sequencing.


Subject(s)
Adult , Female , Humans , Lung Diseases/diagnosis , Mycobacterium/genetics , Mycobacterium Infections/diagnosis , Nontuberculous Mycobacteria/genetics , RNA, Ribosomal, 16S/genetics , Republic of Korea , Sequence Analysis, RNA
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