Bacteremia ; microbiology ; Clostridium Infections ; complications ; Clostridium perfringens ; pathogenicity ; Female ; Hemolysis ; physiology ; Humans ; Sepsis ; microbiology

Anti-Bacterial Agents ; Clostridium Infections ; diagnosis ; microbiology ; Clostridium difficile ; genetics ; Drug Resistance, Bacterial ; genetics ; Humans

BACKGROUND: Antimicrobial susceptibility testing (AST) of Clostridium difficile is increasingly important because of the rise in resistant strains. The standard medium for the AST of C. difficile is supplemented Brucella agar (sBA), but we found that the growth of C. difficile on sBA was not optimal. Because active growth is critical for reliable AST, we developed a new, modified C. difficile (mCD) agar. C. difficile grew better on mCD agar than on sBA. METHODS: C. difficile isolates were collected from patients with healthcare-associated diarrhea. sBA medium was prepared according to the CLSI guidelines. Homemade mCD agar containing taurocholate, L-cysteine hydrochloride, and 7% horse blood was used. For 171 C. difficile isolates, we compared the agar dilution AST results from mCD agar with those from sBA. RESULTS: No significant differences were observed in the 50% minimal inhibitory concentration (MIC50) and 90% minimal inhibitory concentration (MIC90) of clindamycin (CLI), metronidazole (MTZ), moxifloxacin (MXF), piperacillin-tazobactam (PTZ), and rifaximin (RIX), but the values for vancomycin (VAN) were two-fold higher on mCD agar than on sBA. The MICs of CLI, MXF, and RIX were in 100% agreement within two-fold dilutions, but for MTZ, VAN, and PTZ, 13.7%, 0.6%, and 3.1% of the isolates, respectively, were outside the acceptable range. CONCLUSIONS: The MIC ranges, MIC50 and MIC90, were acceptable when AST was performed on mCD agar. Thus, mCD agar could be used as a substitute medium for the AST of C. difficile.
Anti-Infective Agents/*pharmacology ; Clostridium Infections/microbiology ; Clostridium difficile/*drug effects/isolation & purification ; Diarrhea/*microbiology ; Humans ; Microbial Sensitivity Tests/*methods

Clostridium difficile, an anaerobic toxigenic bacterium, causes a severe infectious colitis that leads to significant morbidity and mortality worldwide. Both enhanced bacterial toxins and diminished host immune response contribute to symptomatic disease. C. difficile has been a well-established pathogen in North America and Europe for decades, but is just emerging in Asia. This article reviews the epidemiology, microbiology, pathophysiology, and clinical management of C. difficile. Prompt recognition of C. difficile is necessary to implement appropriate infection control practices.
Asia/epidemiology ; Clostridium Infections/*diagnosis/epidemiology/microbiology/*therapy ; Clostridium difficile/genetics/*pathogenicity ; Europe/epidemiology ; Global Health ; Humans ; North America/epidemiology

We employed a multiplex polymerase chain reaction (PCR) coupled with capillary electrophoresis (mPCR-CE) targeting six Clostridium difficile genes, including tpi, tcdA, tcdB, cdtA, cdtB, and a deletion in tcdC for simultaneous detection and characterization of toxigenic C. difficile directly from fecal specimens. The mPCR-CE had a limit of detection of 10 colony-forming units per reaction with no cross-reactions with other related bacterial genes. Clinical validation was performed on 354 consecutively collected stool specimens from patients with suspected C. difficile infection and 45 isolates. The results were compared with a reference standard combined with BD MAX Cdiff, real-time cell analysis assay (RTCA), and mPCR-CE. The toxigenic C. difficile species were detected in 36 isolates and 45 stool specimens by the mPCR-CE, which provided a positive rate of 20.3% (81/399). The mPCR-CE had a specificity of 97.2% and a sensitivity of 96.0%, which was higher than RTCA (x = 5.67, P = 0.017) but lower than BD MAX Cdiff (P = 0.245). Among the 45 strains, 44 (97.8%) were determined as nonribotype 027 by the mPCR-CE, which was fully agreed with PCR ribotyping. Even though ribotypes 017 (n = 8, 17.8%), 001 (n = 6, 13.3%), and 012 (n = 7, 15.6%) were predominant in this region, ribotype 027 was an important genotype monitored routinely. The mPCR-CE provided an alternative diagnosis tool for the simultaneous detection of toxigenic C. difficile in stool and potentially differentiated between RT027 and non-RT027.
Clostridium Infections ; diagnosis ; Clostridium difficile ; genetics ; Electrophoresis, Capillary ; Feces ; microbiology ; Genes, Bacterial ; Humans ; Polymerase Chain Reaction ; Ribotyping ; Sensitivity and Specificity

Clostridium Infections ; complications ; Fatal Outcome ; Female ; Hemoglobinuria ; etiology ; Hemolysis ; Humans ; Middle Aged ; Sepsis ; complications ; microbiology

Infection with Clostridium difficile is a growing concern because of the increasing prevalence and spread of nosocomial infections. Emergence of the hypervirulent 027/NAP1/BI strain is also notable. Existing diagnostic methods have low sensitivity or are time-consuming. Therefore, establishing a rapid and accurate microbiological diagnostic assay is needed. We evaluated the Xpert C. difficile assay (Xpert CD assay; Cepheid, USA) to detect toxigenic C. difficile. This assay is a real-time multiplex PCR assay that can be used to detect toxigenic C. difficile strains and differentiate the C. difficile presumptive 027/NAP1/BI strain. A total of 253 loose stool specimens were collected and toxigenic cultures, VIDAS C. difficile A & B assays (VIDAS CDAB assay; bioMerieux, France), and the Xpert CD assay were performed. In comparison to toxigenic cultures, the sensitivity, specificity, and positive and negative predictive values were 100%, 94.6%, 83.1%, and 100%, respectively, for the Xpert CD assay and 40.8%, 98.0%, 100%, and 88.9%, respectively, for VIDAS CDAB assay. Because of the low prevalence of the PCR ribotype 027 in Korea, the evaluation of the usefulness of the Xpert CD assay for screening for the 027 strain was limited. The Xpert CD assay provides great sensitivity in diagnosing toxigenic C. difficile infection. In addition, this method has excellent usability because it is simple and fast.
Clostridium Infections/*diagnosis/epidemiology/microbiology ; Clostridium difficile/genetics/*isolation & purification/metabolism ; Face/microbiology ; Humans ; Multiplex Polymerase Chain Reaction ; Prevalence ; Reagent Kits, Diagnostic/*standards ; Sensitivity and Specificity

BACKGROUND: Clostridium difficile is a major cause of antibiotic-associated diarrhea. The objective of this study was to characterize clinical isolates of C. difficile obtained from various regions in Korea with regard to their toxin status, molecular type, and antimicrobial susceptibility. METHODS: We analyzed a total of 408 C. difficile isolates obtained between 2006 and 2008 from 408 patients with diarrhea in 12 South Korean teaching hospitals. C. difficile toxin genes tcdA, tcdB, cdtA, and cdtB were detected by PCR. Molecular genotyping was performed by PCR ribotyping. Antimicrobial susceptibilities of the 120 C. difficile isolates were assessed by agar dilution methods. RESULTS: Among 337 toxigenic isolates, 105 were toxin A-negative and toxin B-positive (A-B+) and 29 were binary toxin-producing strains. PCR ribotyping showed 50 different ribotype patterns. The 5 most frequently occurring ribotypes comprised 62.0% of all identified ribotypes. No isolate was susceptible to cefoxitin, and all except 1 were susceptible to piperacillin and piperacillin-tazobactam. The resistance rates of isolates to imipenem, cefotetan, moxifloxacin, ampicillin, and clindamycin were 25%, 34%, 42%, 51%, and 60%, respectively. The isolates showed no resistance to metronidazole or vancomycin. CONCLUSIONS: This is the first nationwide study on the toxin status, including PCR ribotyping and antimicrobial resistance, of C. difficile isolates in Korea. The prevalence of A-B+ strains was 25.7%, much higher than that reported from other countries. Binary toxin-producing strains accounted for 7.1% of all strains, which was not rare in Korea. The most prevalent ribotype was ribotype 017, and all A-B+ strains showed this pattern. We did not isolate strains with decreased susceptibility to metronidazole or vancomycin.
Clostridium Infections/microbiology ; Clostridium difficile/classification/*genetics/isolation & purification ; Diarrhea/microbiology ; *Drug Resistance, Bacterial ; Enterotoxins/*genetics ; Genetic Variation ; Genotype ; Hospitals, University ; Humans ; Microbial Sensitivity Tests ; Republic of Korea ; Ribotyping

Clostridial septicemia usually occurrs in patients with immunocompromised diseases such as diabetes and malignancy. Clostridial liver abscess is very rare but highly fatal. We experienced a case of Clostridial septicemia due to liver abscess in a 73-year-old man. He was presented with fever and chills. On admission, abdominal CT scan showed about 35 mm sized hypoattenuated lesion with multiple central air-bubbles. After the diagnosis of liver abscess, the patient underwent prompt empirical antimicrobial therapy and percutaneous drainage. In spite of early therapy, the patient had gone into shock and death.
Aged ; Clostridium/*isolation & purification ; Clostridium Infections/diagnosis/*microbiology ; Drainage ; Humans ; Liver/radiography ; Liver Abscess/complications/*diagnosis/microbiology ; Male ; Sepsis/complications/*diagnosis ; Tomography, X-Ray Computed

Pigbel remains a likely significant cause of morbidity and mortality in the highlands of Papua New Guinea (PNG), two decades after the administration of pigbel vaccination ceased. There is a need for an effective surveillance program for pigbel to better understand the disease burden and to target communities for preventive strategies. This paper reviews the epidemiology, pathogenesis, recent history and current data on the burden of pigbel in PNG. We propose a surveillance program based on clinical recognition of likely cases and laboratory confirmation using an ELISA assay for Clostridium perfringens type C beta-toxin. Research aimed at validating this approach in the clinical setting is outlined.
Clostridium Infections/epidemiology/*microbiology/*prevention & control ; Clostridium perfringens/pathogenicity ; Enteritis/epidemiology/*microbiology/*prevention & control ; Health Services Needs and Demand ; Humans ; Incidence ; Papua New Guinea/epidemiology ; Population Surveillance