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*

Objective: To establish and optimize PCR methods for the gene encoding of Clostridium perfringens β₂ toxin (cpb₂) and atypical-cpb₂ (aty-cpb₂), analyze the epidemiological characteristics and genetic polymorphism of the cpb₂ of Clostridium perfringens in 9 Chinese areas from 2016 to 2021. Methods: The cpb₂ of 188 Clostridium perfringens strains were examined by PCR; the cpb₂ sequences were acquired by whole-genome sequencing to analyze the genetic polymorphism. Using Mega 11 and the Makeblastdb tool, a phylogenetic tree, and cpb₂-library based on 110 strains carrying the cpb₂ were produced. Using the Blastn technique, a comparison was made to discover sequence similarity between consensus-cpb₂ (con-cpb₂) and aty-cpb₂. Results: The specificity of PCR assay for the cpb₂ and aty-cpb₂ was verified. The PCR results for cpb₂ amplification were highly consistent with the whole-genome sequencing approach (Kappa=0.946, P<0.001). A total of 107 strains from nine regions in China carried cpb₂, 94 types A strains carried aty-cpb₂, 6 types A strains carried con-cpb₂, and 7 types F strains carried aty-cpb₂. The nucleotide sequence similarity between the two coding genes was 68.97%-70.97%, and the similarity between the same coding genes was 98.00%-100.00%. Conclusions: In this study, a specific PCR method for cpb₂ toxin was developed, and the previous PCR method for detecting aty-cpb₂ was improved. aty-cpb₂ is the primary gene encoding of β₂ toxin. There is a significant nucleotide sequence variance between the various cpb₂ genotypes.
Humans ; Clostridium perfringens/genetics* ; Clostridium Infections ; Bacterial Toxins/genetics* ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Genetic

Clostridioides ; Clostridioides difficile/isolation & purification* ; Clostridium Infections/epidemiology* ; Guidelines as Topic ; Humans

OBJECTIVE: To describe the profile, management, and outcome of adult patients admitted for tetanus. METHODOLOGY: A search of the hospital and department database was conducted for patients with admitting or final diagnosis of tetanus. Patients below 19 years old, with alternative diagnosis, incomplete or unavailable records, transferred to another institution, and went home against medical advice were excluded. Data on patient demographics, source of infection, symptom severity, and management were obtained. Data was described using proportions and averages. RESULTS: Thirty-two patients were included in the study. Twenty-seven were male with a mean age of 45.63 ± 13.39. All cases had no history of tetanus vaccination. The most common focus of infection was acute injuries. More than half of patients would be diagnosed within 72 hours of symptom onset. The most common symptoms on presentation were trismus, rigidity, dysphagia, and spasms. On admission, Cole severity for 11 (34.4%) cases was mild, 17 (53.1%) moderate, and 4 (12.5%) severe. Prophylactic tracheostomy was performed in 31 patients and 19 (59.4%) were placed on assisted ventilation. All cases were treated with metronidazole. A benzodiazepine was started in all cases for spasm control. Baclofen, magnesium sulfate, and antiepileptic drugs such as carbamazepine were also used. Twenty (62.5%) patients suffered from complications, the most common of which was nosocomial pneumonia, which resulted in longer ICU and hospital stays. Eleven patients exhibited progression in Cole severity: 8.6% at stage 1, 57.1% at stage 2, and 34.3% cases were at stage 3. Mortality rate was 19% with the proportion increasing with Cole severity. CONCLUSION We describe the presentation and course of patients admitted for tetanus in our institution. Most cases were males and all cases had no vaccination history. Trismus was the most common presenting symptom. Metronidazole was the antibiotic of choice and benzodiazepines were the mainstay for spasm control. Majority of cases were mild to moderate in severity on presentation. Advanced disease stages were associated with higher fatality rates. Complications were associated with longer ICU and hospital stay. These findings suggest that prevention of progression of disease severity and complications must be the focus of tetanus protocols to shorten hospital stay and decrease mortality rate. Promoting vaccination of at-risk adults is recommended to lower the incidence of tetanus.
Tetanus ; Clostridium tetani ; Clostridium Infections

Intestinal microbiome closely relates with human health and disease, which plays a critical role in the immune response, homeostasis, drug metabolism and tumorigenesis. Imbalances in the composition and function of these intestinal microbes associate with diseases. Fecal microbiota transplantation (FMT) is an established successful treatment modality for recurrent Clostridium difficile infection (CDI). The safety profile and potential therapeutic advantages of FMT for diseases associated with dysbiosis and immune dysfunction have led to many publications, mainly case series. The literature on the use of FMT for hematologic diseases is very limited, however, immune thrombocytopenic purpura（ITP）, CDI and aGVHD after HSCT were reported to be improved by FMT. The aim of this review is to briefly summarize the research current state, procedures and clinical application of FMT.
Clostridium Infections ; Clostridium difficile ; Dysbiosis ; Fecal Microbiota Transplantation ; Hematologic Diseases ; Humans ; Treatment Outcome

Although international clinical guidelines generally recommend bacterial stool cultures for patients with acute diarrhea, stool cultures are frequently being requested by physicians regardless of the likelihood of a bacterial infection. This study was conducted to improve the practice of requesting stool cultures by analyzing patterns for stool culture requests by physicians. We retrospectively reviewed 235 stool cultures of patients who visited Gyeongsang National University Hospital from January to February 2017. We analyzed the period of time after which the stool culture was requested after admission, stool characteristics, wet smear, and concomitant tests performed. 38.7% of stool culture requests were made within 3 days of admission. Stool form analysis showed that 36.6% of stools were watery and loose, and 18.8% were firm. Furthermore, >20 leukocytes per high-power field were found only in 0.4% of the wet smears. Among the stool culture requests, 78.7% were prescribed Clostridium difficile culture or toxin tests at the same time. In addition, 13.6% were prescribed diarrhea-causing viral tests as well. Only stool cultures were requested in 10.2% of the cases. Physicians rarely ensure that the adequate criteria are met when requesting for stool cultures. It is necessary to decrease unnecessary diagnostic practices to maintain the quality of care by establishing reliable rejection criteria and the physicians have valid reasons for requesting stool cultures.
Bacterial Infections ; Clostridium difficile ; Diarrhea ; Humans ; Leukocytes ; Quality Improvement ; Retrospective Studies

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

Fecal microbiota transplantation (FMT) has become a research focus of biomedicine and clinical medicine in recent years. The clinical response from FMT for different diseases provided evidence for microbiota-host interactions associated with various disorders, including Clostridium difficile infection, inflammatory bowel disease, diabetes mellitus, cancer, liver cirrhosis, gut-brain disease and others. To discuss the experiences of using microbes to treat human diseases from ancient China to current era should be important in moving standardized FMT forward and achieving a better future. Here, we review the changing concept of microbiota transplantation from FMT to selective microbiota transplantation, methodology development of FMT and step-up FMT strategy based on literature and state experts' perspectives.
Clostridium Infections ; therapy ; Fecal Microbiota Transplantation ; methods ; standards ; Host Microbial Interactions ; Humans ; Inflammatory Bowel Diseases ; therapy ; Metabolic Diseases ; therapy

PURPOSE: Timely antibiotic therapy in selected cases of diarrhea associated with bacterial infections can reduce the duration and severity of illness and prevent complications. The availability of a predictive index before identification of causative bacteria would aid in the choice of a therapeutic agent. METHODS: The study included patients admitted to the pediatrics unit at Konyang University Hospital for acute inflammatory diarrhea from August 1, 2015 to July 31, 2016 who underwent multiplex polymerase chain reaction testing. Of 248 patients, 83 had positive results. The clinical symptoms and blood test results were examined in 61 patients with Campylobacter spp. (25 patients), Salmonella spp. (18 patients), and Clostridium perfringens (18 patients) infections. The mean age of the 61 patients (male:femal=31:30) was 84.0±54.8 months, and the mean hospital stay was 4.6±1.7 days. RESULTS: There were no statistical differences in sex, age, clinical symptoms, or signs. Patients with Campylobacter infection were significantly older (P=0.00). C-reactive protein (CRP) levels in patients with Campylobacter infection were higher than those in the other 2 groups, at 9.6±6.1 mg/dL. The results of receiver-operating characteristic curve analysis showed that the cutoff age was ≥103.5 months (sensitivity, 72%; specificity, 86%) and the CRP cutoff level was ≥4.55 mg/dL (sensitivity, 80%; specificity, 69%). CONCLUSION: Age (≥103.5 months) and higher CRP level (≥4.55 mg/dL) were good predictors of Campylobacter enterocolitis. If neither criterion was met, Campylobacter enterocolitis was unlikely (negative predictive value 97.2%). When both criteria were met, Campylobacter enterocolitis was highly likely.
Bacteria ; Bacterial Infections ; C-Reactive Protein ; Campylobacter Infections ; Campylobacter* ; Child* ; Clostridium perfringens ; Diarrhea ; Enteritis* ; Enterocolitis ; Hematologic Tests ; Humans ; Length of Stay ; Multiplex Polymerase Chain Reaction ; Pediatrics ; Salmonella ; Sensitivity and Specificity

BACKGROUND/AIMS: Clostridium difficile infection (CDI) has been reported to be a cause of flare-ups in patients with ulcerative colitis (UC). We evaluated the prevalence and clinical outcomes of CDI in patients with UC hospitalized for flare-ups. METHODS: This was a prospective, multicenter study including 7 academic teaching hospitals in Korea. All consecutive patients with UC admitted for disease flare-up were enrolled. We detected the presence of CDI by using enzyme immunoassay, real-time polymerase chain reaction (RT-PCR) for toxin genes, and sigmoidoscopy. RESULTS: Eighty-one consecutive patients with UC were enrolled from January 2014 to December 2015. Among 81 patients, 8 (9.9%) were diagnosed with CDI. Most of the cases were identified by RT-PCR. Enzyme immunoassay was positive in 3 of 8 patients, and only 1 had typical endoscopic findings of pseudomembranous colitis. There were no differences in demographic data, length of hospital stay, or colectomy rate between patients with and without CDI. CONCLUSIONS: CDI was not a rare cause of flare-up in patients with UC in Korea. However, CDI did not appear to affect the course of UC flare-up in Korean patients. RT-PCR was sensitive in detecting CDI and can be considered a diagnostic tool in patients with UC flare-up.
Clostridium difficile* ; Clostridium Infections ; Clostridium* ; Colectomy ; Colitis, Ulcerative* ; Enterocolitis, Pseudomembranous ; Hospitals, Teaching ; Humans ; Immunoenzyme Techniques ; Korea* ; Length of Stay ; Polymerase Chain Reaction ; Prevalence ; Prospective Studies* ; Real-Time Polymerase Chain Reaction ; Sigmoidoscopy ; Ulcer*