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Evaluation of the Usefulness of MacConkey Agar and Colistin-Nalidixic Acid Blood Agar for Body Fluids, Peritoneal Fluid, and Wound/Abscess Specimens.

Jayoung KIM ; Sung Il CHO ; Yong Kyun KIM ; Yeon Joon PARK

Annals of Clinical Microbiology.2015;18(1):1-6. doi:10.5145/ACM.2015.18.1.1

BACKGROUND: Most clinical microbiology laboratories in Korea have difficulty in following the recommendations of the clinical procedure handbook for culture of body fluid and wound/abscess specimens. We evaluated the usefulness of MacConkey (MAC) and colistin-nalidixic acid blood agar (CNA) for the isolation of pathogens from these specimens. METHODS: A total of 1,508 clinical specimens [144 peritoneal fluid, 241 body fluids (19 bile, 70 joint fluid, 6 pericardial fluid, 104 pleural fluid, and other fluids in 42 cases) and 1,123 wound/abscess] were inoculated onto basic media [Blood agar plate (BAP), chocolate agar or BAP with streaking of Staphylococcus aureus] and simultaneously inoculated onto MAC and CNA. The pathogens isolated by basic media and by additional use of MAC and/or CNA were compared. RESULTS: With basic media, 885 isolates from 588 specimens were detected, and by additional use of MAC and CNA, an additional 27 isolates from 24 specimens and an additional 128 isolates from 112 specimens were isolated, respectively. Compared to the basic media, by adding MAC, an additional 233.3%, 38.5% and 4.5% of gram-negative bacteria were isolated from peritoneal fluids, body fluid and wound/abscess, respectively, and by adding CNA, an additional 106.7%, 45.0%, and 20.7% of gram-positive bacteria/ yeast were isolated, respectively. The isolates detected by additional use of MAC were mainly Enterobacteriaceae (77.0%), and those detected by CNA were S. aureus (21.1%), Coagulase-negative Staphylococcus spp. (20.3%), Enterococcus spp. (16.4%), Streptococcus spp. (10.2%) and yeasts (16.4%). CONCLUSION: For peritoneal fluid and body fluid specimens, additional use of MAC plus CNA seems necessary for detection of pathogens. For wound/abscess, additional use of CNA will be cost effective.
Agar* ; Ascitic Fluid* ; Bile ; Body Fluids* ; Cacao ; Enterobacteriaceae ; Enterococcus ; Gram-Negative Bacteria ; Joints ; Korea ; Staphylococcus ; Streptococcus ; Yeasts

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Rapid Diagnosis of Mycobacterium abscessus Bacteremia Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry.

Eun Jeong WON ; Yong Jun CHOI ; Soo Hyun KIM ; Jong Hee SHIN

Annals of Clinical Microbiology.2016;19(3):77-81. doi:10.5145/ACM.2016.19.3.77

Mycobacterium abscessus was isolated from cultures of seven blood samples from a 64-year-old diabetic female who was admitted due to steroid-unresponsive adrenal insufficiency. The isolates were difficult to identify using the conventional commercial systems, VITEK 2 (bioMérieux, France) or MicroScan (Siemens Healthcare Diagnostics, USA), but were rapidly identified as M. abscessus by a matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based Bruker Biotyper system (Bruker Daltonics, USA). Identification of M. abscessus was confirmed by a reverse hybridizationbased assay (Genotype Mycobacterium CM/AS 12, Hain Lifescience) and direct sequencing of a heatshock protein gene. After removal of her central venous catheter, the patient was successfully treated with a combination therapy comprising clarithromycin, amikacin, cefoxitin, and imipenem. Our findings demonstrate that MALDI-TOF MS can facilitate rapid and accurate identification of M. abscessus from blood cultures, which enables prompt administration of appropriate therapy following catheter removal.
Adrenal Insufficiency ; Amikacin ; Bacteremia* ; Catheters ; Cefoxitin ; Central Venous Catheters ; Clarithromycin ; Delivery of Health Care ; Diagnosis* ; Female ; Humans ; Imipenem ; Mass Spectrometry* ; Middle Aged ; Mycobacterium*

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Effects of Blood Volume Monitoring on the Rate of Positive Blood Cultures from the Emergency Room.

Min Kyung SO ; Hae Sun CHUNG ; Chung Jong KIM ; Hee Jung CHOI ; Miae LEE

Annals of Clinical Microbiology.2016;19(3):70-76. doi:10.5145/ACM.2016.19.3.70

BACKGROUND: Blood cultures are essential in diagnosing and treating sepsis. There are several factors that affect the diagnostic yield of blood cultures such as the number of blood sampling episodes, the incubation period, the type and volume of culture media, and the amount of blood drawn. This study aimed to elucidate whether monitoring the volume of blood drawn with an educational intervention could affect the diagnostic quality of blood cultures. METHODS: We implemented quality monitoring for the blood volume drawn during blood culture testing for adults in an emergency room. We instructed the nurses in the emergency room to draw the optimal amount of blood and to reduce the number of blood culture sets from three to two. We analyzed and compared the amount of blood drawn, the rate of positive blood cultures, the contamination rate, and time to positivity (TTP) between 908 patients pre-intervention and 921 patients post-intervention. RESULTS: The amount of blood drawn increased from 0.7±0.3 mL per bottle (pre-intervention) to 6.5±1.7 mL per bottle (post-intervention) (P<0.0001). The rate of positive blood culture post-intervention (12.14%) was higher than that pre-intervention (6.65%) (P<0.0001). The contamination rate post-intervention (1.82%) was also significantly greater than that pre-intervention (0.60%) (P<0.0001). Except for anaerobes, there was no significant difference in the distribution of microorganisms between the pre- and post-intervention periods. The TTP for anaerobe bottles post-intervention was significantly shorter than that of pre-intervention (16.1±16.3 versus 18.6±18.3 h). CONCLUSION: This study suggests that continuing education about adequate blood volume and aseptic techniques is needed to increase the rate of positive blood cultures and reduce the contamination rate of blood cultures.
Adult ; Blood Volume* ; Culture Media ; Education, Continuing ; Emergencies* ; Emergency Service, Hospital* ; Humans ; Sepsis

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Application of Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry to Screen the Extended-Spectrum β-Lactamase-Producing ST131 Escherichia coli Strains.

Young Ah KIM ; Dongeun YONG ; Yong Ha IN ; Hyung Soon PARK ; Kyungwon LEE

Annals of Clinical Microbiology.2016;19(3):65-69. doi:10.5145/ACM.2016.19.3.65

BACKGROUND: Sequence type 131 (ST131) O25b serogroup Escherichia coli, producing CTX-M type extended-spectrum β-lactamase (ESBL), is a major clone involved in worldwide pandemic spread in both community- and healthcare-associated infections. Recently, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has become a routine tool for the identification of bacteria in many laboratories. This study aimed to assess the performance of MALDI-TOF MS for the screening of ESBL-producing E. coli ST131 in a rapid, inexpensive, and simple way. METHODS: A total 26 clinical E. coli, isolated from blood between 2013 and 2014, were used. The characteristics are ST131-O25b ESBL producers (n=6), ST131-O16 ESBL producers (n=4), non-ST131 ESBL producers (n=11), and non-ST131 non-ESBL producers (n=5). Specific biomarker peaks to distinguish the ST131 clonal group from others were investigated by MicroIDSys (ASTA, South Korea) and ASTA Tinkerbell 2.0 software. RESULTS: A peak at 9,713 m/z peak is useful to screen for ST131 E. coli, regardless of serogroup O25 or O16, showing a sensitivity of 100%, specificity of 56.2%, positive predictive value of 58.8%, and negative predictive value of 100% when using a relative intensity cutoff of 15%. CONCLUSION: We can screen for ST131 E. coli using MicroIDSys (ASTA), MALDI-TOF MS in a rapid, inexpensive, and simple way. However, other confirmatory tests are needed to confirm ST131 E. coli due to the low specificity of this method.
Bacteria ; Clone Cells ; Escherichia coli* ; Escherichia* ; Mass Screening ; Mass Spectrometry* ; Methods ; Pandemics ; Sensitivity and Specificity ; Serogroup

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Evaluation of Diagnostic Performance of RAPIDEC CARBA NP Test for Carbapenemase-Producing Enterobacteriaceae.

Wonkeun SONG ; Gilsung YOO ; Gyu Yul HWANG ; Young UH

Annals of Clinical Microbiology.2016;19(3):59-64. doi:10.5145/ACM.2016.19.3.59

BACKGROUND: Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae are resistant to most β-lactam antibiotics except carbapenems. In recent years, infrequently isolated Enterobacteriaceae that produce carbapenemase pose a serious threat in the selection of appropriate therapeutic antibiotics. The rapid detection method of carbapenemase-producing Enterobacteriaceae (CPE) is necessary to prevent the spread of CPE into healthcare facilities. METHODS: One hundred clinical Enterobacteriaceae isolates (Klebsiella pneumoniae 40, Escherichia coli 40, others 20) showing susceptibility to carbapenems and positivity in the CLSI ESBL phenotypic test from November 2015 to March 2016 and 59 stocked Enterobacteriaceae isolates harboring resistance genes producing carbapenemase (K. pneumoniae 56, Enterobacter cloacae 2, E. coli 1; types of CPE: KPC 36, GES 12, NDM 6, VIM 2, OXA 2, IMP 1) were subjected to the RAPIDEC CARBA NP test (bioMérieux, France) and CPE phenotypic test using the modified Hodge test (MHT) and carbapenemase inhibition test. RESULTS: All of the 100 Enterobacteriaceae isolates with carbapenem susceptibility and ESBL positivity were negative on the RAPIDEC CARBA NP test and CPE phenotypic test. Of 59 stock CPE isolates, 53 and 42 showed positive results to the RAPIDEC CARBA NP test and MHT, respectively. The sensitivity and specificity of the RAPIDEC CARBA NP test for detecting CPE were 89.8% and 100%, respectively. CONCLUSION: The RAPIDEC CARBA NP test is simple and produces a result within 3 hr. In conclusion, the test is a useful screen for detecting CPE because it shows high sensitivity and specificity for CPE detection.
Anti-Bacterial Agents ; Carbapenems ; Delivery of Health Care ; Enterobacter cloacae ; Enterobacteriaceae* ; Escherichia coli ; Methods ; Pneumonia ; Sensitivity and Specificity

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Tuberculosis Presenting as a Mediastinal Mass in an Infant.

Namhee KIM ; Hyun Ji LEE ; Jongyoun YI ; Su Eun PARK ; Chulhun L CHANG

Annals of Clinical Microbiology.2017;20(1):17-20. doi:10.5145/ACM.2017.20.1.17

Tuberculosis can occur in various organ systems and may present with diverse manifestations. We report an unusual case of mediastinal tuberculoma in a 3-month-old boy who presented to the hospital after experiencing fever, cough, and progressive pneumonia for two weeks. The chest computed tomography scan indicated a mediastinal mass suggesting lymphoma. However, histological analysis confirmed that the mass was caused by tuberculosis. The present report describes the delayed diagnosis of a disease due to an uncommon presentation. Misdiagnosing unusual cases of tuberculosis results in treatment delays and may lead to an increase in morbidity. Therefore, we suggest that tuberculosis should be included in the differential diagnosis for children presenting with a mediastinal mass, especially in areas with a high prevalence of tuberculosis.
Child ; Cough ; Delayed Diagnosis ; Diagnosis, Differential ; Fever ; Humans ; Infant* ; Lymphoma ; Male ; Mediastinum ; Pneumonia ; Prevalence ; Thorax ; Tuberculoma ; Tuberculosis*

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A Case of Chronic Gordonia otitidis Lung Infection Initially Regarded as Nontuberculous Mycobacterial Lung Disease.

Hyerim KIM ; Yun Seong KIM ; Kye Hyung KIM ; Namhee KIM ; Hyung Hoi KIM ; Chulhun L CHANG ; Jongyoun YI

Annals of Clinical Microbiology.2017;20(1):13-16. doi:10.5145/ACM.2017.20.1.13

The genus Gordonia is one of the mycolic acid-containing aerobic actinomycetes. This genus has 38 named species that are widespread in the natural environment; however, Gordonia species rarely cause human infections. A 76-year-old woman presented with cough and sputum for over 1 year and was suspected of having nontuberculous mycobacterial (NTM) lung disease. An NTM isolate from the sputum was initially identified as Mycobacterium lentiflavum or Mycobacterium genavense by genotypic identification targeting internal transcribed spacer (ITS). However, the isolate was finally confirmed as Gordonia otitidis by sequencing of 16S rRNA, gyrB and secA1 genes. In patients with suspected NTM lung disease, the etiologic agent might be an organism other than NTM such as G. otitidis but still be identified as NTM without sequencing of 16S rRNA or other genes. Especially in case that a possible NTM isolate is identified as M. lentiflavum or M. genavense by the genotypic method targeting ITS, additional genotypic tests such as sequencing of 16S rRNA and other genes would be necessary for more reliable identification.
Actinobacteria ; Actinomycetales ; Aged ; Cough ; Female ; Humans ; Lung Diseases* ; Lung* ; Methods ; Mycobacterium ; Nontuberculous Mycobacteria ; Respiratory Tract Infections ; Sputum

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Effect of Delayed Entry on Time to Detection for the Lactose Nonfermentative Gram-Negative Rods.

Seung Wook KIM ; Jung Hyun BYUN ; Sunjoo KIM

Annals of Clinical Microbiology.2017;20(1):7-12. doi:10.5145/ACM.2017.20.1.7

BACKGROUND: Prolonged transport or poor accessibility of blood culture equipment during night time may cause delayed entry of blood culture bottles. The effect of prestorage conditions on time to detection (TTD) for the blood culture was evaluated for the important gram-negative lactose nonfermentative bacteria. METHODS: Three different clinical isolates of Pseudomonas aeruginosa, Acinetobacter baumannii, Stenotrophomonas maltophilia, and Burkholdera cepacia were diluted to 150 CFU/mL and 15 CFU/mL and inoculated into standard aerobic bottles. These were stored at 25℃ and at 37℃ for 0, 6, 12, 18, and 24 h. They were entered to BacT/Alert 3D Systems (bio-Mérieux Inc.) and TTD was monitored for each condition. RESULTS: At the 150 CFU/mL concentration, P. aeruginosa and A. baumannii showed false-negative for the bottles prestored at 37℃ for 18 h and 24 h, respectively. However, there was no false-negative for S. maltophilia or B. cepacia at any prestorage conditions. There was a significant decrease of TTD for all experimental microorganisms except P. aeruginosa prestored for 24 h either at 25℃ or at 37℃ (P< 0.05). CONCLUSION: Delayed entry may cause false-negative, especially for the high level of bacteremia of P. aeruginosa or A. baumannii when the bottles are stored at 37℃ for ≥18 h. TTD could be reduced by prestorage of the bottles at 37℃ until 12 h without false-negative for nonfermentative bacteria.
Acinetobacter baumannii ; Bacteremia ; Bacteria ; Lactose* ; Pseudomonas ; Pseudomonas aeruginosa ; Sepsis ; Stenotrophomonas maltophilia

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Comparative Evaluation of Multiplex Real-Time PCR Assays for Six Pathogens of Sexually Transmitted Infections.

Hae Sun CHUNG ; Miae LEE

Annals of Clinical Microbiology.2017;20(1):1-6. doi:10.5145/ACM.2017.20.1.1

BACKGROUND: The multiplex real-time PCR assay is a sensitive test for simultaneous detection of various pathogens of sexually transmitted infections (STIs). We evaluated the performance of two multiplex real-time PCR assays for six STI pathogens. METHODS: DNA samples after being used to conduct PCR for STI pathogens were stored below −70℃. Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), Mycoplasma genitalium (MG), Mycoplasma hominis (MH), Ureaplasma urealyticum (UU), and Trichomonas vaginalis (TV) were detected by multiplex real-time PCR with GeneFinder STD I (CT/NG/UU)/II (MG/MH/TV) Multiplex Real-time PCR Kits (Infopia, Korea; GeneFinder assay) and Real-Q CT&NG/MH&TV/MG&UU Kits (BioSewoom, Korea; Real-Q assay). Discrepant results were resolved by another multiplex real-time assay, Anyplex II STI-7 Detection (Seegene, Korea). Any two positive results for the assays were considered true positive. RESULTS: Among 81 samples, the GeneFinder assay detected 63 pathogens from 45 cases (16 CT, 2 NG, 6 MG, 20 MH, 18 UU, and 1 TV) and Real-Q assay detected 66 pathogens from 47 cases (16 CT, 2 NG, 8 MG, 20 MH, 19 UU, and 1 TV). For the results of positive cases and negative cases, the overall concordance rate between the two multiplex real-time assays was 93.8% (Kappa=0.87). For each pathogen, the agreement rates of the two assays ranged from 97.5 to 100% (Kappa>0.8). CONCLUSION: There was no significant difference between the results of GeneFinder assay and Real-Q assay. Both multiplex real-time PCR assays can be useful methods for the detection of STI pathogens in clinical laboratories.
Chlamydia trachomatis ; DNA ; Korea ; Mycoplasma genitalium ; Mycoplasma hominis ; Neisseria gonorrhoeae ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction* ; Sexually Transmitted Diseases* ; Trichomonas vaginalis ; Ureaplasma urealyticum

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A Case of Misidentification of Aspergillus versicolor Complex as Scopulariopsis Species Isolated from a Homograft.

Hee Jae HUH ; Jang Ho LEE ; Kyung Sun PARK ; Tae Gook JUN ; I Seok KANG ; Yae Jean KIM ; Chang Seok KI ; Nam Yong LEE

Annals of Clinical Microbiology.2013;16(2):105-109. doi:10.5145/ACM.2013.16.2.105

We report a case of the isolation of the Aspergillus versicolor complex, initially misidentified by morphological characteristics as the Scopulariopsis species, from a homograft with a bicuspidalized pulmonary valve. An eighteen-month-old female, who had critical pulmonary stenosis, underwent pulmonary valve replacement. On postoperative day 8, she developed a fever, which did not respond to empiric broad-spectrum antibiotics. While no definitive source was identified, a filamentous fungus was isolated from the thawed homograft tissue culture prior to implantation on the operation day. The colonies were powdery green with white edges on Sabouraud dextrose agar. Microscopic examination showed septate hyphae with branched conidiophores and chains of spiny conidia, which suggested Scopulariopsis species. After direct sequencing of the internal transcribed spacer (ITS) regions, the fungus was identified as the A. versicolor complex. To our knowledge, the isolation of the A. versicolor complex from a homograft valve has not been previously described. This case shows that laboratory staff should be aware that microscopic morphology of the A. versicolor complex can resemble that of a number of other genera, including Scopulariopsis species.
Agar ; Anti-Bacterial Agents ; Aspergillus ; Bicuspid ; European Continental Ancestry Group ; Female ; Fever ; Fungi ; Glucose ; Humans ; Hyphae ; Pulmonary Valve ; Pulmonary Valve Stenosis ; Scopulariopsis ; Spores, Fungal ; Transplantation, Homologous

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