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

Clostridium perfringens can produce many kinds of toxins and hydrolase, causing gas gangrene, enteritis and enterotoxemia in both human and animals. It is known that C. perfringens can produce more than 20 toxins and hydrolases. The different toxin types are associated with specific disease types. At present, molecular toxin-typing method by PCR has replaced the traditional serological typing method. In this study, we systematically summarize the types, basic characteristics, pathogenic mechanism and the relationship with disease of C. perfringens toxins to provide evidence for the establishment of rapid detection method, immune antigen screening, antibody preparation and research of related pathogenic mechanism.
Animals ; Humans ; Clostridium perfringens ; Antibodies ; Polymerase Chain Reaction

BACKGROUND: We attempted to determine the characteristics of diarrheal pathogens according to species, seasonal variations, and patient age using multiplex PCR for the epidemiologic study of diarrheal disease in Jeju Island. METHODS: From March 2015 to Feb 2017, stool specimens were collected from 537 diarrheal patients older than 16 years. Multiplex PCR was used to identify pathogens and found Group A Rotavirus, enteric Adenovirus, Norovirus GI/GII, Astrovirus, Salmonella spp., Shigella spp., Vibrio spp., Campylobacter spp., Clostridium difficile toxin B (CDB), Clostridium perfringens, Yersinia enterocolitica, Aeromonas spp., Escherichia coli O157:H7, and verocytotoxin-producing E. coli (VTEC). RESULTS: Pathogens were isolated from 221 of 537 samples (41.2%); 9.3% were positive only for viral pathogens; 30.2%, only for bacterial pathogens; and 1.7%, for both viral and bacterial pathogens. Bacteria were more prevalent in spring, summer, and autumn, but viral pathogens were more prevalent in winter. Overall prevalence were Campylobacter spp. (26.7%), Clostridium perfringens (23.9%); Norovirus GII (11.4%), CDB (8.2%), Aeromonas spp. (6.3%), Group A Rotavirus (5.1%), Salmonella spp. (3.9%), Astrovirus (3.9%), Norovirus GI (3.1%), Vibrio spp (2.7%), enteric Adenovirus (1.6%), Shigella spp. (1.2%), VTEC (1.2%), and Yersinia enterocolitica (0.4%). Group A Rotavirus and Norovirus GII were more prevalent in winter and early spring. Campylobacter spp., CDB, and C. perfringens were detected frequently, without seasonal variation. CONCLUSION: Bacterial pathogens are more prevalent than viruses in acute diarrhea in adults living in Jeju Island, especially in spring, summer, and autumn. Viral pathogens are prevalent in winter. Campylobacter spp., CDB, and Clostridium perfringens are the major pathogens occurring without seasonal variations. These data will be helpful in identifying diarrheal pathogens and for treatments and prevention strategies.
Adenoviridae ; Adult* ; Aeromonas ; Bacteria ; Campylobacter ; Clostridium difficile ; Clostridium perfringens ; Diarrhea ; Epidemiologic Studies* ; Epidemiology ; Escherichia coli ; Humans ; Multiplex Polymerase Chain Reaction* ; Norovirus ; Prevalence ; Rotavirus ; Salmonella ; Seasons ; Shiga-Toxigenic Escherichia coli ; Shigella ; Vibrio ; Yersinia enterocolitica

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

Clostridium perfringens causes diarrhea and other diseases in animals and humans. We investigated the prevalence, toxin gene profiles, and antibiotic resistance of C. perfringens isolated from diarrheic dogs (DD) and non-diarrheic dogs (ND) in two animal hospitals in Seoul, Korea. Fecal samples were collected from clinically DD (n = 49) and ND (n = 34). C. perfringens was isolated from 31 of 49 DD (63.3%) and 21 of 34 ND dogs (61.8%). All C. perfringens strains were positive for the α toxin gene, but not for the β, ε, or ι toxin genes; therefore, all strains were identified as type A C. perfringens. All isolates were cpe-negative, whereas the β2 toxin gene was identified in 83.9% and 61.9% of isolates from DD and ND, respectively. Most isolates were susceptible to ampicillin (94%), chloramphenicol (92%), metronidazole (100%), moxifloxacin (96%), and imipenem (100%). However, 25.0% and 21.2% of isolates were resistant to tetracycline and clindamycin, respectively. Molecular subtyping of the isolated strains was performed by using pulsed-field gel electrophoresis. Fifty-two isolates were classified into 48 pulsotypes based on more than 90% similarity of banding patterns. No notable differences were observed among the isolates from DD and ND.
Ampicillin ; Animals ; Bacterial Toxins ; Chloramphenicol ; Clindamycin ; Clostridium perfringens ; Clostridium ; Diarrhea ; Dogs ; Drug Resistance ; Drug Resistance, Microbial ; Electrophoresis, Gel, Pulsed-Field ; Hospitals, Animal ; Humans ; Imipenem ; Korea ; Metronidazole ; Prevalence ; Seoul ; Tetracycline

Gas gangrene, a subset of necrotizing myositis, is a bacterial infection that produces gas in tissues in gangrene. It is usually caused by Clostridium species, most commonly Clostridium perfringens. Streptococcus anginosus is a rare cause of gas gangrene, with very few cases reported. We report a rare case of traumatic gas gangrene caused by S. anginosus in a 57-year-old female with diabetes after being stabbed with scissors.
Bacterial Infections ; Clostridium ; Clostridium perfringens ; Diabetes Mellitus ; Female ; Gangrene ; Gas Gangrene* ; Humans ; Middle Aged ; Myositis ; Streptococcus anginosus* ; Streptococcus*

A 65-year-old female visited the emergency room for severe back pain radiating to the neck. Aortic dissection computed tomography revealed a ruptured liver abscess and large pneumoperitoneum. Although emergent percutaneous drainage of the liver abscess and aggressive resuscitation were performed, massive hemolytic anemia and disseminated intravascular hemolysis developed and she subsequently died, 11 hours after her visit to the emergency room. Clostridium perfringens was identified in a blood culture obtained at the emergency room. We report this case because refractory septic shock due to a liver abscess and massive intravascular hemolytic anemia caused by Clostridium perfringens in a healthy female is rare.
Aged ; Anemia, Hemolytic* ; Back Pain ; Clostridium perfringens* ; Clostridium* ; Disseminated Intravascular Coagulation ; Drainage ; Emergency Service, Hospital ; Female ; Hemolysis ; Humans ; Liver Abscess* ; Liver* ; Neck ; Pneumoperitoneum ; Resuscitation ; Shock, Septic*

BACKGROUND: The rapid and accurate detection of diarrheal pathogens is essential to prevent the spread of diarrheal diseases. Recently, a multiplex PCR assay was developed to simultaneously detect various bacterial and viral diarrheal pathogens. In this study, we investigated the frequency of detection of various potential pathogens causing diarrhea by using multiplex PCR and compared the results to the results of stool culture tests for bacteria and enzyme immunoassays (EIAs) for rotaviruses and Clostridium difficile toxin B (CDTB). METHODS: We retrospectively analysed the results for multiplex PCR, culture tests, and EIA obtained from stool specimens submitted to the laboratory from May 2013 to September 2014. Multiplex PCR was performed using the Seeplex diarrhea ACE detection kit (Seegene, Korea), which detects five viruses and eight bacteria. RESULTS: Among 890 stool specimens, 408 (45.8%) were found to be positive by PCR. The PCR positivity rate for bacteria and viruses was 31.1% (277/890) and 18.9% (161/890), respectively. The relative frequencies of microorganisms or toxins detected by PCR were, in decreasing order, CDTB 24.0%, Clostridium perfringens 20.6%, norovirus-GII 15.8%, rotavirus 11.3%, Campylobacter spp. 7.5%, enteric adenovirus 5.7%, and Salmonella spp. 5.1%. The concordance rate of the results obtained using the PCR and culture tests was 99.2% for Salmonella spp., 95.7% for Campylobacter spp., and. 79.8% for C. difficile . The concordance rates for rotaviruses and CDTB were 99.7% and 83.6%, respectively. CONCLUSIONS: The multiplex PCR method showed a high detection rate and is useful for the simultaneous detection of various diarrheal pathogens.
Adenoviridae ; Bacteria ; Campylobacter ; Clostridium difficile ; Clostridium perfringens ; Diarrhea ; Immunoenzyme Techniques ; Multiplex Polymerase Chain Reaction* ; Polymerase Chain Reaction ; Retrospective Studies ; Rotavirus ; Salmonella

Pleuropulmonary diseases caused by Clostridial species infections are rare, but have a mortality rate of up to 30%. Furthermore, older people are at greater risk of developing invasive clostridium infections, and the majority of reported cases of clostridium empyema have been attributed to iatrogenic trauma or aspiration. The authors report a case of spontaneous empyema caused by Clostridium perfringens. A 72-year-old woman was admitted to Kangwon National University Hospital for empyema. The patient had no history of trauma, a dental procedure, or aspiration, and was treated using empirical antibiotics and by drainage of pleural fluid. Bacteria species that cause empyema are usually not detected, but on the 4th day of admission, C. perfringens was isolated from the pleural space. The patient was continuously treated with antibiotics for C. perfringens and drainage, and was discharged 25 days after admission with almost a fully recovered status. Increased awareness of Clostrium species infection in the elderly is needed to ensure appropriate treatment.
Aged ; Anti-Bacterial Agents ; Bacteria ; Clostridium ; Clostridium Infections ; Clostridium perfringens* ; Drainage ; Empyema* ; Female ; Gangwon-do ; Humans ; Mortality

Necrotic enteritis (NE) caused by Clostridium (C.) perfringens commonly occurs in domestic broiler farms since antibiotic supplementation in poultry feed has been banned. We evaluated the antibacterial activities of medicinal plant extracts against C. perfringens isolates to select alternative compounds for preventing NE. We compared antibacterial activities using two methods and evaluated susceptibilities of the isolates based on minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Two (Fraxinus rhynchophylla Hance [FRH] and Geranium koreanum Kom. [GKK]) of the 30 plant extracts had potent antibacterial activities against C. perfringens ATCC 13124 in two assays. The MIC values for FRH and GKK against 20 C. perfringens isolates were 128~256 microg/mL and 32~128 microg/mL, respectively. The geometric MIC mean values for the two extracts were 147.2 microg/mL and 68.8 microg/mL, respectively. The MBCs for the two extracts against the same strains were 1,024~2,048 microg/mL and 256~1,024 microg/mL, respectively. The geometric mean MIC and MBC for GKK were about two-fold lower than those of FRH. The modified spot-on-lawn assay may be useful for measuring primary antibacterial potential. FRH and GKK are expected to be used as feed additives to prevent or treat NE in veterinary practice.
Chickens* ; Clostridium ; Clostridium perfringens* ; Enteritis ; Fraxinus* ; Geranium* ; Microbial Sensitivity Tests ; Plant Extracts ; Plants, Medicinal ; Poultry