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Detection of Chlamydophila pneumoniae in Acute Myocardial Infarction.

Won Kil LEE ; Eun Hee KWON ; Hye Gyung BAE ; Jang Soo SUH ; Kyung Eun SONG ; Nan Young LEE ; Dong Il WON ; Jung Bum LEE

Korean Journal of Clinical Microbiology.2003;6(1):81-87.

BACKGROUND: There is growing evidence linking infection with Chlamydophila pneumoniae with vascular diseases, such as atherosclerosis and myocardial infarction. However, the data remain inconclusive and the clinical importance of C. pneumoniae as vasculopathic is unclear. So, we intend to detect C. pneumoniae in acute myocardial infarction patients by microimmunofluorescence (mIF) and polymerase chain reaction (PCR). METHODS: Blood and peripheral mononuclear cells (PMNCs) of 24 myocardial infarction patients and 100 normal controls were collected. Serum were used in mIF and PMNCs in PCR. PMNC sample were tested for C. pneumoniae by 'touchdown 'nested PCR. The first round PCR amplified DNA from both C. pneumoniae and Chlamydophila psittaci, while the second round specially targeted C. pneumoniae allowing the two species to be differentiated. RESULTS: Seropositivity of IgG and IgM anti-Chlamydophila pneumoniae antibody titers were 95.8% and 25% in myocardial infarction patients and 61% and 16% in control group, respectively. Positive rates of PCR of PMNCs were 8.3% in the patients and 15% in control group. CONCLUSION: The results of mIF show that mIF positive rate in myocardial infarction was much higher than control group. So an association between C. pneumoniae and myocardial infarction can be concluded. But the opposite results of PCR of PMNCs needed further studies.
Atherosclerosis ; Chlamydial Pneumonia* ; Chlamydophila pneumoniae* ; Chlamydophila psittaci ; Chlamydophila* ; DNA ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Myocardial Infarction* ; Pneumonia ; Polymerase Chain Reaction ; Vascular Diseases

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Detection of Ureaplasma urealyticum and Mycoplasma hominis in Pregnant Women Using MYCOFAST(R) Evolution 2 and PCR.

Hye Gyung BAE ; Woon Bo HEO ; Nang Young LEE ; Won Kil LEE ; Tae Bon KOO

Korean Journal of Clinical Microbiology.2003;6(1):74-80.

BACKGROUND: The associations between preterm labor or premature rupture of membrane (PROM) and urogenital infections of pregnant women are reported. Ureaplasma urealyticum and Mycoplasma hominis are well known as important pathogens of urogenital infections in pregnant women. In routine clinical laboratory, conventional culture for these microorganisms has not been made generally because of the requirements for strict growth condition. MYCOFAST(R) Evolution 2 is an easy and rapid liquid microculture method using metabolism of these microorganisms. Author investigated the relationship between U. urealyticum or M. hominis infections and preterm labor or PROM by MYCOFAST Evolution 2 and PCR. Also it was reviewed that the possibility of substitution of MYCOFAST Evolution 2 for conventional culture method by comparing with PCR methods. METHODS: This study was done on 91 pregnant women. They were composed of two groups; group I(n=48) had full-term delivery and group II(n=43) had preterm labor or PROM before the 37th week.Two cervical swabs were made each time. One was used for MYCOFAST(R) Evolution 2 and the other for PCR. RESULTS: The positivity of U. urealyticum was 39.6% in group Iand 58.1% in group IIby MYCOFAST Evolution 2 and 39.6% and 58.1% by PCR method, respectively. The positivity of M. hominis was 4.2% in group Iand 11.6% in group IIby MYCOFAST Evolution 2 and 4.2% and 7.0% by PCR method, respectively. The positivity of U. urealyticum and M. hominis in group IIwas higher than that in group Ibut was not significant statistically. The concordance rates between two methods were 86.8% for U. urealyticum and 97.8% for M. hominis. It showed good correlation between two methods (U. urealyticum, r=0.736; M. hominis, r=0.835). CONCLUSIONS: The infections of U. urealyticum and M. hominis were related to preterm labor or PROM. Considering vertical transmission to fetus or neonates resulting in perinatal morbidity or mortality, the detection of these microorganisms is important. MYCOFAST(R) Evolution 2 was an easy, rapid and reliable method substituting conventional culture method.
Female ; Fetus ; Humans ; Infant, Newborn ; Membranes ; Metabolism ; Mortality ; Mycoplasma hominis* ; Mycoplasma* ; Obstetric Labor, Premature ; Polymerase Chain Reaction* ; Pregnancy ; Pregnant Women* ; Rupture ; Ureaplasma urealyticum* ; Ureaplasma*

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Evaluation of MGIT 960 System for Recovery of Mycobacteria from Body Fluids.

Youn Mi CHOI

Korean Journal of Clinical Microbiology.2003;6(1):69-73.

BACKGROUND: In this study, we evaluated the BACTEC MGIT 960 system (Becton Dickinson Microbiology Systems, Sparks, Md, USA), which is fully automated, noninvasive and nonradiometric fluorescent indicator broth detection system, for the growth and detection of mycobacteria with body fluid specimens. METHODS: Total of 1,891 body fluid specimens were included (pleural fluid 752, ascitic fluid 629, cerebrospinal fluid 214, joint fluid 79, peritozol 54, others 163). Specimens were inoculated into MGIT and solid media (3% ogawa, Japan). Polymerase chain reaction was performed for the discrimination of Mycobacterium tuberculosis from Mycobacterium other than tuberculosis (MOTT). RESULTS: A total of 62 isolates of mycobacteria were recovered from all culture system. With MGIT system, 56 isolates were recovered, compared with solid system recovered 33 isolates. 29 isolates were recovered with MGIT only and 6 isolates recovered with solid media only. Among 62 isolates recovered, 11 isolates were positive in acid fast stain. 10 isolates were recovered with MGIT. One isolate was recovered with solid system. 51 isolates were negative in acid fast stain. Among this, 46 isolates were recovered with MGIT. The mean detection time was 14.2 days with MGIT system, and 38.2 days with solid media. Contamination rate for each system with body fluid specimens were 4.1% for MGIT and 1.7% for solid media. CONCLUSION: In body fluid, the MGIT system has the advantages of improved detection rate and rapid recovery than solid media to recover mycobacteria.
Ascitic Fluid ; Body Fluids* ; Cerebrospinal Fluid ; Discrimination (Psychology) ; Joints ; Mycobacterium ; Mycobacterium tuberculosis ; Polymerase Chain Reaction ; Tuberculosis

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Direct Detection of Clostridium difficile Toxin B Gene by Nested PCR in Human Stool Specimens.

Hee Kyung PARK ; Young Mi LEE ; Hyun Jung JANG ; Cheol Min KIM ; Kyungwon LEE ; Seok Hoon JEONG ; Mooin PARK

Korean Journal of Clinical Microbiology.2003;6(1):63-68.

BACKGROUND: Clostridium difficile is the major cause of antibiotic-associated diarrhea (AAD) and pseudomembranous colitis (PMC). The aim of this study was to develop the nested PCR assay for direct detection of toxigenic C. difficile in stool specimens and to evaluate the usefulness of the method. METHODS: Specificity of newly designed primers are tested with 36 reference strains of intestinal flora. Lower detection limit of nested PCR for B toxin gene in C. difficile was determined using 10-fold serial dilutions of C. difficile ATCC 9689. One hundred and two clinical stool samples were cultured for detection of C. difficile on cycloserine-cefoxitin- fructose agar and the PCR assay for detection of toxin B gene in C. difficile isolates was performed. Nested PCR assay for direct detection of toxin B gene in clinical samples was also performed. RESULTS: Nested PCR assay showed negative amplification results in intestinal floras except C. difficile ATCC 9689. Lower detection limit of nested PCR for toxin B gene was 10 4 CFU/mL. Sensitivity of nested PCR assay compared to culture method was 100% (29/29), and the specificity was 68% (50/73). CONCLUSION: Nested PCR assay showed high sensitivity in direct detection of toxin B gene in C. difficile isolates even after administration of metronidazole, so the assay could be used in initial diagnosis and follow-up tests of AAD and PMC.
Agar ; Clostridium difficile* ; Clostridium* ; Diagnosis ; Diarrhea ; Enterocolitis, Pseudomembranous ; Follow-Up Studies ; Fructose ; Humans* ; Limit of Detection ; Metronidazole ; Polymerase Chain Reaction* ; Sensitivity and Specificity

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Quality Assurance for Commercially Prepared Microbiological Culture Media.

Tae Yeal CHOI

Korean Journal of Clinical Microbiology.2003;6(1):56-62.

BACKGROUND: Culture media that perform as intended are crucial to accurate work by a clinical microbiology laboratory. The author evaluated commercially prepared microbiological culture media for quality assurance. MATERIAL AND METHODS: Five types of commercially prepared media from Shin Yang Chemical Co., Ltd. were evaluated in Hanyang University Hospital for one year. Five types included blood agar media, chocolate agar, MacConkey agar, Salmonella-Shigella agar and Mueller-Hinton agar. All media were evaluated by NCCLS M22-A2 (Quality assurance for commercially prepared microbiological culture media-second edition; approved standard, 1996). RESULTS: Blood agar media provided luxuriant growth of many bacteria-especially the fastidious streptococci and pneumococci, as shown by early colonial development and clear hemolytic reactions with S. pyogeses. Chocolate agar supported growth of fastidious bacteria such as N. gonorrheae and H. influenzae. MacConkey agar gave excellent differentiation between coliforms and non-lactose fermenters with inhibition of Gram-positive micrococci. Salmonella-Shigella agar was a good differential selective medium for the isolation of Salmonella and some Shigella species from clinical specimens. Mueller-Hinton agar showed good reproducibility for antimicrobial susceptibility test by the disk diffusion method. CONCLUSION: The five types of commercially prepared culture media have demonstrated competence and integrity for bacterial culture in clinical trial. Theses media could be used without performance quality assurance test by user.
Agar ; Bacteria ; Cacao ; Culture Media* ; Diffusion ; Gonorrhea ; Influenza, Human ; Mental Competency ; Quality Control ; Salmonella ; Shigella

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A Prospective, Comparative Study of Two Methods of Ascitic Fluid Culture to Diagnose Spontaneous Bacterial Peritonitis..

In Sook KIM ; Nam Yong LEE

Korean Journal of Clinical Microbiology.2003;6(1):52-55.

BACKGROUND: Inoculation of ascitic fluid into blood culture bottles is known to be more efficient than conventional culture method to diagnose spontaneous bacterial peritonitis. The aim of this study is to evaluate recovery and early detection of peritonitis-causing bacteria from ascitic fluid by using the BACTEC blood culture system with bedside inoculation. The results were compared to those of conventional culture method. METHODS: Ascitic fluid specimens from 345 patients suspicious of spontaneous bacterial peritonitis were prospectively evaluated between January 1999 and March 2002. In all cases, 5 to 10 mL of ascitic fluid were inoculated at the bedside into a pair of BACTEC blood culture bottles (BC method), and simultaneously an aliquot of ascitic fluid was sent to microbiology laboratory for conventional culture. Isolated microorganisms and the time elapsed for final report were compared between the two methods. RESULTS: BC method was positive in 66/345 ascitic fluid specimens (19.1%) and conventional culture method in 48/345 (13.9%) (P=0.065). Time elapsed for final report was 82 20.5 hours for blood culture method and 107 42.4 hours for conventional culture method (P=0.002). CONCLUSION: BC method using BACTEC system provides an earlier microbiologic diagnosis of spontaneous bacterial peritonitis than conventional culture method with higher sensitivity.
Ascitic Fluid* ; Bacteria ; Diagnosis ; Humans ; Peritonitis* ; Prospective Studies*

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Antibiogram of Escherichia coli and Klebsiella spp. Detected by Vitek ESBL Test.

Eun Hae CHO ; Nam Yong LEE

Korean Journal of Clinical Microbiology.2003;6(1):47-51.

BACKGROUND: Extended-spectrum beta -lactamase (ESBL) producing Escherichia coli and Klebsiella spp. isolates are clinically resistant to all the beta -lactams except carapenems and cephamycins. This study was to determine the prevalence of ESBL producing E. coli and Klebsiella spp. and the rates and trends of resistance to extended-spectrum beta -lactams and other antimicrobial agents in ESBL producing E. coli and Klebsiella spp.. METHODS: During the periods of 2002, a total 2,551 clinical isolates of E. coli & Klebsiella spp. were collected from patients of the Samsung medical center, Seoul, Korea. Antimicrobial susceptibility test and determination of ESBL production were performed by Vitek GNS-433 card. RESULTS: 151/1,594 (9.5%) of E. coli isolates, 128/896 (14.3%) of K. pneumoniae isolates and 11/61 (18.0%) of K. oxytica were ESBL producing strains. Resistance to cefoxitin and cefepime were 2.4% and 13.4% in ESBL producing isolates. Imipenem had excellent activity against E .coli and Klebsiella spp. (100% susceptible). CONCLUSION: In this study, ESBL-producing E. coli and Klebsiella spp were more resistant to beta -lactams including cefepime than ESBL non-producing E. coli and Klebsiella spp.. ESBL producing E. coli and Klebsiella spp. showed a high level of co-resistance with aminoglycosides and fluoroquinolones. Imipenem showed the highest level of activity against E. coli and Klebsiella spp..
Aminoglycosides ; Anti-Infective Agents ; Cefoxitin ; Cephamycins ; Escherichia coli* ; Escherichia* ; Fluoroquinolones ; Humans ; Imipenem ; Klebsiella* ; Korea ; Microbial Sensitivity Tests* ; Pneumonia ; Prevalence ; Seoul

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Comparison of MicroScan and Vitek ESBL test with NCCLS ESBL Confirmatory Test.

Myung Hee LEE

Korean Journal of Clinical Microbiology.2003;6(1):41-46.

BACKGROUND: This study was designed to evaluate the ability of the Vitek and MicroScan ESBL test by comparing with NCCLS ESBL phenotypic confirmatory test by disk diffusion and to know the frequency of ESBL producers in the Seoul Veterans Hospital. METHODS: A total of 1,261 isolates(Escherichia coli 705, Klebsiella pneumoniae 502, K. oxytoca 54) from 883 patients were included in ESBL screening test by Vitek (494 strains) and MicroScan (767 strains). After excluding repetitive isolates from same patients, NCCLS ESBL confirmatory test was performed for 197 ESBL screening positives and 184 ESBL screening negatives. RESULTS: The overall frequency of ESBL screening positives was 22.3% (by MicroScan 26.2%, by Vitek 15.6%), and that of NCCLS ESBL positives was 18.9%(18.3% in E. coli, 21% in K. pneumoniae). MicroScan and Vitek ESBL test showed 100% and 92.3% sensitivity, 77.1% and 95.5% specificity, respectively. Among the 158 NCCLS ESBL positives, 17.7% showed clavulanic acid effect in cefotaxime only, 10.1% in ceftazidime only, and 72.2% in both. MicroScan Neg ComboPanel Type 21 test revealed that 91.4% of suspicious ESBL producers flagged by one or two antimicrobials were erroneous. In contrast, 96.2% of strains flagged by all five antimicrobials were correct. CONCLUSION: Suspicious ESBL producers by MicroScan showing three or four antimicrobial flags should be retested by NCCLS ESBLconfirmatory test. But strains with two or less flags and strains with all 5 flags can be reported as Non-ESBL producers and ESBL producers, respectively.
Cefotaxime ; Ceftazidime ; Clavulanic Acid ; Diffusion ; Hospitals, Veterans ; Humans ; Klebsiella pneumoniae ; Mass Screening ; Sensitivity and Specificity ; Seoul

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In Vitro Antimicrobial Activity of Cefroxadine, an Oral Cephalosporin, Against Major Clinical Isolates.

Jongyoun YI ; Jae Kyu LEE ; Eui Chong KIM

Korean Journal of Clinical Microbiology.2003;6(1):37-40.

BACKGROUND: Cefroxadine is an oral first-generation cephalosporin, which has been used for several years. But, the susceptibility data of cefroxadine were rarely reported in Korea. The current study attempted to determine the antibacterial activity of cefroxadine against the major clinical isolates. METHODS: According to the NCCLS recommendations, antibacterial activities of cefroxadine were measured against total 500 major clinical isolates. MICs were determined by the agar dilution method, a series of doubling dilutions from 128 to 0.03 /mL, on Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Citrobacter freundii, Serratia marcescens, Proteus mirabilis, and Staphylococcus spp. In case of Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis, broth microdilution method, a series of doubling dilutions from 16 to 0.015 /mL, was performed. RESULTS: Cefroxadine had variable activity against Enterobacteriaceae. MIC cumulative curves showed that cefroxadine had relatively low MIC distributions against E. coli, K. pneumoniae and P. mirabilis, showing MIC50 were 4, 4, and 8 /mL, respectively. Against E. cloacae, C. freundii, and S. marcescens, cefroxadine 's MIC50 values ranged from 128 to >128 /mL. For clinical isolates of methicillin-susceptible Staphylococcus aureus and methicillin-susceptible Staphylococcus epidermidis, cefroxadine had MIC90 values were 4 /mL and 8 /mL, respectively. Cefroxadine had MIC50 values of 1 /mL and >16 /mL for penicillin-susceptible and penicillin-not-susceptible strains of S. pneumoniae, respectively. Cefroxadine had MIC50 values of 8 /mL and 4 /mL against H. influenzae and M. catarrhalis, respectively. CONCLUSION: Cefroxadine had good activity against gram-positive bacteria, except penicillin-resistant S. pneumoniae, and showed moderate antimicrobial activity against M. catarrhalis, E. coli, P. mirabilis, and K. pneumonaie. Cefroxadine had variable activity against Enterobacteriaceae other than the above-mentioned species.
Agar ; Citrobacter freundii ; Cloaca ; Enterobacter cloacae ; Enterobacteriaceae ; Escherichia coli ; Gram-Positive Bacteria ; Haemophilus influenzae ; Influenza, Human ; Klebsiella pneumoniae ; Korea ; Mirabilis ; Moraxella (Branhamella) catarrhalis ; Pneumonia ; Proteus mirabilis ; Serratia marcescens ; Staphylococcus ; Staphylococcus aureus ; Staphylococcus epidermidis ; Streptococcus pneumoniae

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A Case of Skin and Soft Tissue Infection by Mycobacterium abscessus.

Nam Hee RYOO ; Won Mok LEE ; Jung Sook HA ; Dong Suk JEON ; Jae Ryong KIM ; Dae Gu SOHN ; Shin Woo KIM

Korean Journal of Clinical Microbiology.2005;8(2):198-201.

A 36-year-old female who initially presented with a small erythematous and swollen abscess on her left anterior tibial area was found out to have a cutaneous Mycobacterium abscessus infection. She was first treated with incision and drainage, dressing, and antibiotics. The lesion began to be aggravated and dispersed. Neither aerobic nor anaerobic bacteria was grown on blood agar plate. After a few weeks, Mycobacterium grew on Ogawa media after 6 days, and was identified as M. abscessus by PCR-restriction fragment length polymorphism. She was then treated with clarithromycin, levofloxacin, and amikacin, and the skin lesion was resolved without further recurrence.
Abscess ; Adult ; Agar ; Amikacin ; Anti-Bacterial Agents ; Bacteria, Anaerobic ; Bandages ; Clarithromycin ; Drainage ; Female ; Humans ; Levofloxacin ; Mycobacterium* ; Recurrence ; Skin* ; Soft Tissue Infections*

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