BACKGROUND: As tuberculous and nontuberculous mycobacterial infections are increasing, it is very important to differentiate the myobacterial species. High performance liquid chromatography (HPLC) method has been proven to be a useful technique for the identification of mycobacteria. The purpose of this study was to investigate the identification rate using HPLC and to know nontuberculous mycobacterial distribution in Ulsan University Hospital. METHOD: Mycobacteria grew in 959 clinical specimens, which were analyzed by HPLC, and their distribution was reviewed by retrospective studies. RESULTS: The patterns of HPLC were divided into single, double, and triple cluster groups which consist of 9, 20, and 4 species of mycobacteria respectively. The identification rate of mycobacteria by HPLC was 98.9%, And the rate of nontuberculous mycobacteria in mycobacterial culture positive specimens was 12.2%. CONCLUSION: HPLC is an excellent tool for mycobacterial identification. And the culture rate of nontuberculous mycobacteria in clinical specimens is increasing in Korea.
Chromatography, High Pressure Liquid ; Chromatography, Liquid* ; Korea ; Mycobacterium tuberculosis ; Nontuberculous Mycobacteria ; Retrospective Studies ; Ulsan

BACKGROUND: During the specimen preparation, neutralization after alkali processing is not necessary when inoculate into agar or broth media, but into 3% Ogawa media, the mostly used egg-based media in Korea. To simplify the specimen processing, the recovery of mycobacteria from 3% Ogawa media inoculating differently processed sputa, was evaluated. Futhermore, the recovery from the three different media was evaluated, too. METHODS: 209 sputa were included. Each specimen was divided into 3 bottles.(A) One was only decontaminated by 4% NaOH before inoculation.(B) Another was decontaminated, and concentrated without neutralization. These two specimens were inoculated into 3% Ogawa media.(C) The other was decontaminated, neutralized and concentrated, and innoculated into 3% Ogawa, Middlebrook 7H9 selective broth and 7H10 selective agar media. The isolates were identified by AccuProbe method. RESULTS: M. tuberculosis were isolated from 43(20.5%) specimens. The recovery number and time from Ogawa media, inoculating specimens processed by A, B, and C method, were 29(13.9%), 35(16.7%), and 36(17.2%), and 23, 23, and 19 days, respectively. The recovery number and time from Ogawa, 7H10 agar and 7H9 broth media, inoculating specimens processed by C method, were 36(17.2%), 42(20.1%), and 41(19.6%), and 26, 24, and 22 days, respectively. CONCLUSIONS: Neutralization of alkali-decontaminated specimens could be used to inoculate into 3% Ogawa media. Inoculation into homemade 7H10 agar and 7H9 broth media in addition to 3% Ogawa media improved the recovery rate and time of mycobacteria.
Agar ; Alkalies ; Korea ; Mycobacterium tuberculosis ; Sputum ; Tuberculosis

A long nose with a drooping tip is a major aesthetic problem. It creates a negative and aged appearance and looks worse when smiling. In order to rectify this problem, the underlying anatomical causes should be understood and corrected simultaneously to optimize surgical outcomes. The causes of a drooping tip of a long nose are generally classified into two mechanisms. Static causes usually result from malposition and incorrect innate shape of the nasal structure: the nasal septum, upper and lower lateral cartilages, and the ligaments in between. The dynamic causes result from the facial expression muscles, the depressor septi nasi muscle, and the levator labii superioris alaeque nasi muscle. The depressor septi nasi depresses the nasal tip and the levator labii superioris alaeque nasi pulls the alar base upwards. Many surgical methods have been introduced, but partial approaches to correct such deformities generally do not satisfy East Asians, making the problem more challenging to surgeons. Typically, East Asians have thick nasal tip soft tissue and skin, and a depressed columella and alar bases. The authors suggest that multifactorial approaches to static and dynamic factors along with ancillary causes should be considered for correcting the drooping tip of the long noses of East Asians.
Asian Continental Ancestry Group* ; Cartilage ; Congenital Abnormalities ; Esthetics ; Facial Expression ; Humans ; Ligaments ; Muscles ; Nasal Septum ; Nose* ; Rhinoplasty ; Skin ; Smiling

BACKGROUND: Infections caused by nontuberculous mycobacteria (NTM) are significantly increasing over the last decade. Due to the uncertainty in the clinical significance of these organisms, their effective diagnosis and treatment has been challenging. The purpose of this study was to investigate the distribution and clinical significance of NTM in clinical specimens. METHODS: Acid-fast culture positive 3,107 clinical specimens were identified by mycolic acid analysis using high performance liquid chromatography (HPLC.) The HPLC patterns of 384 NTM strains were compared with those of standard mycobacterium species. Clinical significance of NTM was investigated by a retrospective study including acid-fast stain and culture, medical history, symptoms and signs, radiological and other laboratory findings, pathologic findings, response to treatment, and follow-up study, and was confirmed according to the guideline of American Thoracic Society. RESULTS: Among the 3,107 Mycobacterium-positive specimens, 384 (12.4%) were found to be positive for NTM. Of these, 367 (95.6%) were successfully identified by HPLC as 19 different species, each of which comprising 0.3% to 15.9% of the total NTM, Studies on the pathogenic role of NTM showed that 0~79.6% of each species or 0~100% of isolates from each specimen could be considered clinically significant. CONCLUSION: HPLC method is highly discriminative for the identification of NTM in clinical specimens. When NTM is isolated from clinical specimens in the Ulsan area, the findings from this study could serve as a database on which to determine its clinical significance depending on species type and also specimen type.
Chromatography, High Pressure Liquid ; Chromatography, Liquid ; Follow-Up Studies ; Mycobacterium ; Mycolic Acids ; Nontuberculous Mycobacteria ; Retrospective Studies ; Uncertainty

BACKGROUND: Infections caused by mycobacteria have been significantly increasing. Due to the difficulty of making a decision about the pathogenicity of mycobacteria, species-level identification is very important for patients' diagnosis and treatment. The purpose of this study was to identify mycobacteria species using a high performance liquid chromatography (HPLC) method and to provide an initial database for the distribution of mycobacteria in Korea. METHODS: Acid fast bacteria isolated from 3,107 clinical specimens were identified by mycolic acid analysis using HPLC. The HPLC patterns were compared with those of standard mycobacteria species. RESULTS: The HPLC patterns were divided into single, double, and triple cluster groups, each group comprising 9, 20, and 4 species, respectively. Mycobacteria and non-tuberculous mycobacteria (NTM) were identifies by HPLC at the rates of 99.5% and 95.6%, respectively. NTM was isolated in 12.4% of the mycobacteria positive specimens. This study also found that there were 20 different NTM species with the distribution of each species ranging from 0.3% to 15.9% of the total NTM. While the rate of NTM has been increasing in Korea, M. avium-intracellulare, M. fortuitum, and M. chelonae are relatively decreasing, and M. kansasii and M. gordonae are relatively increasing. CONCLUSIONS: HPLC method was highly discriminative for the identification of NTM in clinical specimens.
Bacterial Typing Techniques ; Chromatography, High Pressure Liquid/*methods ; Hospitals, University ; Humans ; Korea ; Mycobacteria, Atypical/chemistry/*isolation & purification ; Mycobacterium Infections, Atypical/drug therapy/microbiology ; Mycolic Acids/analysis

BACKGROUND: The Amplified Mycobacterium tuberculosis Direct Test (AMTDT) has been developed for the direct detection of M. tuberculosis complex in respiratory specimens. Traditional methods for diagnosis of extrapulmonary tuberculosis such as the acid-fast bacilli (AFB) stain have their well-known limitations. We investigated the usefulness of the AMTDT for a wide range of non-respiratory specimens to establish early diagnosis of extrapulmonary tuberculosis. METHODS: 346 specimens (219 urine, 117 pleural fluid, 6 ascitic fluid, 2 lymph node, 1 gastric aspirate, and 1 pus specimens) from 340 patients referred from November 1997 to September 1998 were tested by the AMTDT. The AMTDT results were evaluated by comparing with clinical diagnosis and smear results. RESULTS: The overall sensitivity, specificity, and positive and negative predictive values of the AMTDT were 82.9%, 93.8%, 64.2%, and 97.6%, respectively. There were no difference in sensitivity and specificity between pleural fluid and urine specimens. In 31 specimens from tuberculosis patients concurrently tested with AMTDT and stain, 15 were only AMTDT positive and 4 were only stain positive. Among the results considered to be false positive, 47.2% of cases were shown as being less than 150,000 relative light units (RLU). In 30 specimens from tuberculosis patients during or after treatment, all six of the patients with reactivation or aggravation were AMTDT positive, and one case was considered to be false positive. CONCLUSIONS: Our study demonstrates the efficacy of the AMTDT in diagnosing extrapulmonary tuberculosis. Prudent interpretation of the AMTDT's results is recommended in case of that being less than 150,000 RLU.
Ascitic Fluid ; Diagnosis ; Early Diagnosis ; Humans ; Lymph Nodes ; Mycobacterium tuberculosis* ; Mycobacterium* ; Polymerase Chain Reaction ; Sensitivity and Specificity ; Suppuration ; Tuberculosis

Neonatal hemochromatosis is a very rare disorder with an unclear etiology with an extremely poor outcome. We report a case of female newborn who presented with direct hyperbilirubinemia without any evidence of congenital infection, immune related hemolysis or exogenous iron uptake. Diagnostic studies revealed highly elevated ferritin level. T2 weighted image of abdominal Magnetic Resonance Imaging (MRI) showed decreased signal intensity of entire left lobe and a part of right lobe of liver with normal spleen. Liver biopsy showed a large amount of hemosiderin in the hepatocytes and Kupffer cells. All the biochemical and excretory liver functions normalized with conservative managements.
Biopsy ; Female ; Ferritins ; Hemochromatosis* ; Hemolysis ; Hemosiderin ; Hepatocytes ; Humans ; Hyperbilirubinemia ; Infant, Newborn ; Iron ; Kupffer Cells ; Liver ; Magnetic Resonance Imaging ; Spleen

The timely detection of blood-borne pathogens is one of the most important functions of the microbiology laboratory. Recently, methicillin-resistant staphylococci have become the most important pathogens seen by the laboratory. The purpose of this study was to evaluate Staphy agar, a novel screening medium, for the detection methicillin-resistant Staphylococcus aureus, S. epidermidis, or other coagulase-negative staphylococci (CNS) from positive blood cultures showing Gram-positive cocci in clusters. Eighty-six blood cultures that yielded Gram-positive cocci in clusters were included in this study. The organisms were finally identified by the Vitek system, and oxacillin resistance was confirmed by polymerase chain reaction (PCR)-based mecA gene detection. The identification and oxacillin resistance of all S. aureus strains showed complete agreement with the Vitek and PCR results. The presumptive detection of S. epidermidis and other CNS were consistent with the Vitek system in 94.7%, and the screening of oxacillin resistance was consistent with the result of PCR in 92.1% of 38 strains. The Staphy agar method is reliable and rapid for differentiating Gram-positive cocci in clusters in blood and for determining their methicillin resistance.
*Bacterial Proteins ; Carrier Proteins/genetics ; Drug Resistance, Microbial ; *Hexosyltransferases ; Muramoylpentapeptide Carboxypeptidase/genetics ; Oxacillin/*pharmacology ; Penicillin-Binding Proteins ; *Peptidyl Transferases ; Staphylococcus aureus/*drug effects/genetics ; Staphylococcus epidermidis/*drug effects/genetics

BACKGROUND: The isolation of carbapenemase-producing Enterobacteriaceae (CPE) has become increasingly common. Continuous surveillance for these organisms is essential because their infections are closely related to outbreaks of illness and are associated with high mortality rates. The aim of this study was to develop and evaluate multiplex PCR as a means of detecting several important CPE genes simultaneously. METHODS: We aimed to develop a multiplex PCR that could detect seven CPE genes simultaneously. The multiplex PCR was composed of seven primer sets for the detection of KPC, IMP, VIM, NDM-1, GES, OXA-23, and OXA-48. We designed different PCR product sizes of at least 100 bp. We evaluated the performance of this new test using 69 CPE-positive clinical isolates. Also, we confirmed the specificity to rule out false-positive reactions by using 71 carbapenem-susceptible clinical strains. RESULTS: A total of 69 CPE clinical isolates showed positive results and were correctly identified as KPC (N=14), IMP (N=13), OXA-23 (N=12), OXA-48 (N=11), VIM (N=9), GES (N=5), and NDM (N=5) by the multiplex PCR. All 71 carbapenem-susceptible clinical isolates, including Enterococcus faecalis , Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, showed negative results. CONCLUSION: This multiplex PCR can detect seven CPE genes at a time and will be useful in clinical laboratories.
Acinetobacter baumannii ; Disease Outbreaks ; Enterobacteriaceae ; Enterococcus faecalis ; Escherichia coli ; Klebsiella pneumoniae ; Mortality ; Multiplex Polymerase Chain Reaction ; Polymerase Chain Reaction ; Pseudomonas aeruginosa ; Sensitivity and Specificity

BACKGROUND: Mycobacterial disease is still greatly concerned in the developing and industrialized countries. Ogawa media has been used to diagnose mycobacterial disease in Korea in spite of a low sensitivity and long incubation time. Mycobacterium Growth Indicator Tube (MGIT) 960 system has been developed to overcome the pitfalls of Ogawa media. So, we investigated improvement in dectection rate and the detection time of mycobacteria using the MGIT 960 system along with 3% Ogawa media. METHODS: A total of 8,045 clinical specimens referred to the department of laboratory medicine in Ulsan University Hospital from January in 2001 to June in 2002 were cultured for mycobacteria. Specimens were processed with the NALC-NAOH (final concentration of NaOH: 1%) and inoculated into both MGIT and Ogawa media. Mycolic acid in the cultured products were analyzed by High performance liquid chromatography to discriminate between Mycobacterium tuberculosis and nontuberculous mycobacteria. RESULTS: Of 8,045 clinical specimens cultured, mycobacteria grew in 957 (11.9%) specimens, 840 (87.8%) M. tuberculosis and 117 (12.2%) nontuberculous mycobacteria. Mycobacteria were detected in 939 specimens (98.1%) by MGIT and 771 (80.6%) specimens by Ogawa media; 753 (78.7%) were detected by both media, 186 (19.4%) by MGIT only, and 18 (1.9%) by Ogawa media only. Mycobacteria were detected in 11.7 days by MGIT 960 and 28.4 days by Ogawa media. CONCLUSIONS: The detection rate and detection time of mycobacteri are improved considerably by the MGIT system; however a combined use of MGIT system and Ogawa media is the most ideally recommended for increasing the detection rate and shortening the detection time.
Chromatography, Liquid ; Developed Countries ; Korea ; Mycobacterium tuberculosis ; Mycobacterium* ; Mycolic Acids ; Nontuberculous Mycobacteria ; Tuberculosis ; Ulsan