Objective: We analyze the characteristics of Clostridioides difficile (C. difficile) infection among diarrhea patients in Kunming from 2018 to 2020 and provide evidence for follow-up surveillance and prevention. Methods: A total of 388 fecal samples of diarrhea patients from four sentinel hospitals in Yunnan Province from 2018 to 2020 were collected. Real-time quantitative PCR was used to detect the fecal toxin genes of C. difficile. The positive fecal samples isolated the bacteria, and isolates were identified by mass spectrometry. The genomic DNA of the strains was extracted for multi-locus sequence typing (MLST). The fecal toxin, strain isolation, and clinical patient characteristics, including co-infection with other pathogens, were analyzed. Results: Among the 388 fecal samples, 47 samples with positive reference genes of C. difficile were positive, with a total positive rate of 12.11%. There were 4 (8.51%) non-toxigenic and 43 (91.49%) toxigenic ones. A total of 18 strains C. difficile were isolated from 47 positive specimens, and the isolation rate of positive specimens was 38.30%. Among them, 14 strains were positive for tcdA, tcdB, tcdC, tcdR, and tcdE. All 18 strains of C. difficile were negative for binary toxins. The MLST results showed 10 sequence types (ST), including 5 strains of ST37, accounting for 27.78%; 2 strains of ST129, ST3, ST54, and ST2, respectively; and 1 strain of ST35, ST532, ST48, ST27, and ST39, respectively. Fecal toxin gene positive (tcdB+) results were statistically associated with the patient's age group and with or without fever before the visit; positive isolates were only statistically associated with the patient's age group. In addition, some C. difficile patients have co-infection with other diarrhea-related viruses. Conclusions: The infection of C. difficile in diarrhea patients in Kunming is mostly toxigenic strains, and the high diversity of strains was identified using the MLST method. Therefore, the surveillance and prevention of C. difficile should be strengthened.
Humans ; Bacterial Toxins/genetics* ; Enterotoxins/genetics* ; Clostridioides difficile/genetics* ; Multilocus Sequence Typing ; Coinfection ; Bacterial Proteins/genetics* ; China/epidemiology* ; Clostridium Infections/epidemiology* ; Diarrhea/microbiology*

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

BACKGROUND: We evaluated the performance of four commercial nucleic acid amplification tests (NAATs: Xpert C. difficile, BD MAX Cdiff, IMDx C. difficile for Abbott m2000, and Illumigene C. difficile) for direct and rapid detection of Clostridium difficile toxin genes. METHODS: We compared four NAATs on the same set of 339 stool specimens (303 prospective and 36 retrospective specimens) with toxigenic culture (TC). RESULTS: Concordance rate among four NAATs was 90.3% (306/339). Based on TC results, the sensitivity and specificity were 90.0% and 92.9% for Xpert; 86.3% and 89.3% for Max; 84.3% and 94.4% for IMDx; and 82.4% and 93.7% for Illumigene, respectively. For 306 concordant cases, there were 11 TC-negative/NAATs co-positive cases and 6 TC-positive/NAATs co-negative cases. Among 33 discordant cases, 18 were only single positive in each NAAT (Xpert, 1; Max, 12; IMDx, 1; Illumigene, 4). Positivity rates of the four NAATs were associated with those of semi-quantitative cultures, which were maximized in grade 3 (>100 colony-forming unit [CFU]) compared with grade 1 (<10 CFU). CONCLUSIONS: Commercial NAATs may be rapid and reliable methods for direct detection of tcdA and/or tcdB in stool specimens compared with TC. Some differences in the sensitivity of the NAATs may partly depend on the number of toxigenic C. difficile in stool specimens.
Bacterial Proteins/genetics ; Bacterial Toxins/genetics ; Clostridium Infections/*diagnosis/microbiology ; Clostridium difficile/*genetics/isolation & purification ; DNA, Bacterial/*analysis/metabolism ; Enterotoxins/genetics ; Feces/*microbiology ; Humans ; *Multiplex Polymerase Chain Reaction ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

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

OBJECTIVEClostridium difficile is an obligate anaerobic Gram-positive bacillus, it can cause Clostridium difficile-associated diarrhea (CDAD). This study aimed to investigate the virulence genes and clinical features of CDAD in children by gene detection.

METHODFrom May 2012 to January 2013, the 121 inpatients in Beijing Children's Hospital who suffered from diarrhea after antibiotics treatment were detected for Clostridium difficile virulence genes including the five genes for pathogenic loci (tcdA, tcdB, tcdC, tcdD, tcdE) and the genes for binary toxin CDT (cdtA and cdtB) using polymerase chain reaction (PCR) in order to research the clinical features of CDAD, and analyze target products by sequencing.

RESULTSIn the 121 children with diarrhea, 60 (49.6%) were toxin B-positive,including 12 toxin A-positive and toxin B-positive (A+B+), 48 toxin A-negative but toxin B-positive (A-B+). The toxin A-positive but toxin B-negative (A+B-) specimens or binary toxin (cdtA and cdtB)-positive specimens were not detected. Of 60 tcdB-positive specimens, tcdC, tcdD and tcdE positive specimens were 24 (40%), 25 (42%), 24 (40%), respectively. The sequencing results of tcdA, tcdB, tcdC, tcdD, and tcdE gene were consistent with the reference sequence. In the 60 children with CDAD, infants (≤3 years) accounted for 62% (37/60). The duration of diarrhea was 3-77 days, and 42 (70%) cases had acute diarrhea; 39 (65%) patients had fever, 40 (67%) had anemia, 36 (60%) had abnormal white blood cell count, 30 (50%) had hypoalbuminemia, 25 (42%) had elevated C-reactive protein (CRP). The level of CRP in positive group was significantly higher compared to the negative group (45.0(16.0,89.0) mg/L vs. 19.0(14.5,41.5) mg/L, Z=-2.008, P=0.045). The level of plasma albumin in positive group was significantly lower compared to the negative group (35.3(29.7,39.8) g/L vs. 38.5(33.9,41.5) g/L, Z=-2.610, P=0.009). There were no significant differences in gender, age, duration of diarrhea, hospital staytime, time of using antibiotics and laboratory test between A+B+ group and A-B+ group (all P>0.05).

CONCLUSIONThe main virulence genotype of Clostridium difficile was toxin A-negative but toxin B-positive in this research. The clinical features of CDAD in children were acute diarrhea with fever. Laboratory examination showed that white blood cell count was abnormal, CRP was increased, hemoglobin and plasma albumin were reduced.

Anti-Bacterial Agents ; Bacterial Toxins ; genetics ; Beijing ; C-Reactive Protein ; Child ; Clostridium Infections ; microbiology ; Clostridium difficile ; genetics ; pathogenicity ; Diarrhea ; microbiology ; Fever ; Genes, Bacterial ; Genotype ; Humans ; Infant ; Leukocyte Count ; Polymerase Chain Reaction ; Virulence ; genetics

OBJECTIVETo analyze the incidence and treatment of Clostridium difficile infection (CDI) in pediatric patients with antibiotic-associated diarrhea (AAD).

METHODClinical data of totally 577 pediatric patients with AAD seen from January 2012 to January 2014 were collected; those children were divided according to age into 4 groups, 0.25 -1 year, >1 -4 years, >4 -12 years and >12 -18 years old groups, and 220 healthy children were enrolled as controls. CDI was tested by C. Diff Quik Chek Complete (QCC) and BD GeneOhm™ C. Diff Assay (BD-PCR) in all children, and the CDI incidence of four groups was added up. All pediatric patients with AAD were divided into mild, general and severe type according to different symptoms of diarrhea, and grading treatment, the general type and severe type of CDI children were treated with metronidazole and (or) vancomycin, afterwards, the results of grading treatment were analyzed.

RESULTThe number of pediatric patients with AAD were 178, 177, 132 and 90 in 0. 25 - 1 year, > 1 -4 years, > 4 - 12 years and > 12 - 18 years old group, respectively. The positive rate of CDI (22. 0% (39/177)) in > 1 -4 years old AAD patients was very significantly higher compared to the controls (4% (4/91), P < 0. 001), the rate of CDI (21. 2% (28/132)) in > 4 - 12 years old AAD pediatric patients was significantly higher compared to the controls (4% (2/53), P = 0. 004), the rates of CDI in 0. 25 - 1 year and > 12 - 18years old AAD groups were not significantly different from that of the controls (P >0. 05). There were 285 mild type AAD children (no CDI children), 176 general type AAD children (including 47 CDI children), and 116 severe type AAD children (including 81 CDI children). After grading and symptomatic treatment, there were 16 recurrent diarrhea in 128 CDI patients (severe type AAD), and the rest recovered. Two cases were transferred for referral treatment, 2 cases died, and the rest 12 recurrent diarrhea children fully recovered after administration of metronidazole, vancomycin, probiotics and symptomatic treatment.

CONCLUSIONThe > 1 -12 years old AAD children had higher CDI rate than healthy children; administration of metronidazole and (or) vancomycin was effective for CD infection.

Adolescent ; Anti-Bacterial Agents ; therapeutic use ; Case-Control Studies ; Child ; Child, Preschool ; Clostridium Infections ; drug therapy ; Clostridium difficile ; Diarrhea ; microbiology ; Humans ; Incidence ; Metronidazole ; therapeutic use ; Probiotics ; therapeutic use ; Vancomycin ; therapeutic use

Hepatocellular carcinoma (HCC) is a critical global health issue and the third most common cause of cancer-related deaths worldwide. The majority of patients who present HCC are already at an advanced stage and their tumors are unresectable. Sorafenib is a multi-kinase inhibitor of the vascular endothelial growth factor pathway and was recently introduced as a therapy for advanced HCC. Furthermore, studies have shown that oral sorafenib has beneficial effects on survival. However, many patients experience diverse side effects, and some of these are severe. Liver abscess development has not been previously documented to be associated with sorafenib administration in HCC. Here, we report the case of a HCC patient that developed a liver abscess while being treated with sorafenib.
Anti-Bacterial Agents/therapeutic use ; Antineoplastic Agents/adverse effects/*therapeutic use ; Carcinoma, Hepatocellular/*drug therapy/radiography ; Clostridium/isolation & purification ; Clostridium Infections/drug therapy/microbiology ; Humans ; Liver Abscess/etiology/*microbiology ; Liver Neoplasms/*drug therapy/radiography ; Male ; Middle Aged ; Niacinamide/adverse effects/*analogs & derivatives/therapeutic use ; Phenylurea Compounds/adverse effects/*therapeutic use ; Tomography, X-Ray Computed

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

Clostridium (C.) difficile is a common cause of nosocomial diarrhea in horses. Vancomycin and metronidazole have been used as standard treatments but are only moderately effective, which highlights the need for a novel alternative therapy. In the current study, we prepared antiserum of equine origin against both C. difficile toxins A and B as well as whole-cell bacteria. The toxin-neutralizing activities of the antibodies were evaluated in vitro and the prophylactic effects of in vivo passive immunotherapy were demonstrated using a conventional mouse model. The data demonstrated that immunized horses generated antibodies against both toxins A and B that possessed toxin-neutralizing activity. Additionally, mice treated with the antiserum lost less weight without any sign of illness and regained weight back to a normal range more rapidly compared to the control group when challenged orally with 10(7) C. difficile spores 1 day after serum injection. These results indicate that intravenous delivery of hyperimmune serum can protect animals from C. difficile challenge in a dose-dependent manner. Hence, immunotherapy may be a promising prophylactic strategy for preventing C. difficile infection in horses.
Animals ; Antibodies, Bacterial/blood/*immunology/therapeutic use ; Bacterial Proteins/immunology/therapeutic use ; Bacterial Toxins/immunology/therapeutic use ; Clostridium Infections/microbiology/prevention & control/*veterinary ; Clostridium difficile/*immunology ; Enterotoxins/immunology/therapeutic use ; Female ; Horse Diseases/microbiology/*prevention & control ; Horses ; Immune Sera/*immunology ; Immunization, Passive/*veterinary ; Mice ; Mice, Inbred C57BL ; Spores, Bacterial/immunology