Antibiotic susceptibility and genome analysis of Enterococcus species isolated from inpatients in one hospital with no apparent outbreak of vancomycin-resistant Enterococcus in Japan.

To prevent nosocomial infection, it is important to screen for potential vancomycin-resistant Enterococcus (VRE) among patients. In this study, we analyzed enterococcal isolates from inpatients in one hospital without any apparent outbreak of VRE. Enterococcal isolates were collected from inpatients at Hiroshima University Hospital from April 1 to June 30, 2021 using selective medium for Enterococci. Multilocus sequence typing, antimicrobial susceptibility testing, and whole-genome sequencing were performed. A total of 164 isolates, including Enterococcus faecium (41 isolates), Enterococcus faecalis (80 isolates), Enterococcus raffinosus (11 isolates), Enterococcus casseliflavus (nine isolates), Enterococcus avium (12 isolates), Enterococcus lactis (eight isolates), Enterococcus gallinarum (two isolates), and Enterococcus malodoratus (one isolate), were analyzed. We found one vanA-positive E. faecium, which was already informed when the patient was transferred to the hospital, nine vanC-positive E. casseliflavus, and two vanC-positive E. gallinarum. E. faecium isolates showed resistance to ampicillin (95.1%), imipenem (95.1%), and levofloxacin (87.8%), and E. faecalis isolates showed resistance to minocycline (49.4%). Ampicillin- and levofloxacin-resistant E. faecium had multiple mutations in penicillin-binding protein 5 (PBP5) (39/39 isolates) and ParC/GyrA (21/36 isolates), respectively. E. raffinosus showed resistance to ampicillin (81.8%), imipenem (45.5%), and levofloxacin (45.5%), and E. lactis showed resistance to ampicillin (37.5%) and imipenem (50.0%). The linezolid resistance genes optrA and cfr(B) were found only in one isolate of E. faecalis and E. raffinosus, respectively. This study, showing the status of enterococci infection in hospitalized patients, is one of the important information when considering nosocomial infection control of VRE.

First reported human case of isolation of Vagococcus fluvialis from the urine of a former zoo clerk in Japan: a case report.

BACKGROUND: Vagococcal infections are extremely rare in humans. There are limited studies on the optimal methods for identification, antimicrobial susceptibility testing, and clinical manifestations of vagococcal infections. Herein, we report a patient with a urinary tract infection who had Vagococcus fluvialis in the urine. CASE PRESENTATION: An 84-year-old man presented to our urology department with a fever that had persisted for several days. He previously worked as a zoo clerk. The patient underwent a left nephroureterectomy for ureteral cancer 5 years ago, and total cystectomy and right cutaneous ureterostomy for muscle-invasive bladder cancer 1 year prior. He was empirically treated with 500 mg of levofloxacin intravenously every 24 h for the urinary tract infection. V. fluvialis was detected in his urine samples and Pseudomonas aeruginosa was detected in his urine and blood samples. Two bacterial species were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. He was administered intravenous levofloxacin for approximately 1 week, followed by oral levofloxacin for another week, after which the infections were eradicated. CONCLUSIONS: To the best of our knowledge, this is the first report of V. fluvialis detected in human urine in Japan. Vagococcus spp. is commonly isolated from fish or animals, and based on the patient's work history, it is possible that the patient was a carrier because of transmission from animals.

A case of bacteremia caused by Dialister micraerophilus with Enterocloster clostridioformis and Eggerthella lenta in a patient with pyometra.

BACKGROUND: Infection by Dialister micraerophilus, an obligate anaerobic gram-negative bacillus, has rarely been described, and its clinical characteristics remain unclear. CASE PRESENTATION: We report a case of bacteremia caused by D. micraerophilus, Enterocloster clostridioformis, and Eggerthella lenta in a 47-year-old woman, associated with pyometra. D. micraerophilus was identified using 16S rRNA gene sequencing and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. D. micraerophilus was detected by polymerase chain reaction using D. micraerophilus-specific primers and E. clostridioformis and E. lenta was isolated from the drainage pus sample obtained from the pyometra uterus. The patient achieved a cure after abscess drainage and 2-week antibiotic treatment. CONCLUSIONS: To the best of our knowledge, this is the first report of D. micraerophilus bacteremia. D. micraerophilus may be associated with gynecological infections. Clinicians should consider both oral and gynecological sites when searching to identify the focus of D. micraerophilus infection.

Complete genome sequence of cfr(B)-carrying Enterococcus raffinosus isolated from bile in a patient in Japan.

OBJECTIVES: Linezolid is an antibiotic used to treat infectious diseases caused by vancomycin-resistant enterococci and methicillin-resistant Staphylococcus aureus. Recently, Enterococcus Spp.-carrying mobile linezolid resistance genes were reported. Herein, we report the complete genome sequence of Enterococcus raffinosus JARB-HU0741, which was isolated from a bile sample of a patient in Japan on May 5, 2021, and carries a linezolid resistance gene, cfr(B). Nevertheless, this isolate was susceptible to linezolid. METHODS: Whole-genome sequencing was performed using HiSeq X FIVE (Illumina) and GridION (Oxford Nanopore Technologies). The sequence reads were assembled using Unicycler v0.4.8, and the complete genome was annotated using DFAST v1.2.18. Antimicrobial resistance genes were detected with Abricate v1.0.1, using the ResFinder database. The minimum inhibitory concentrations (MICs) were determined using broth microdilution and interpreted according to the guidelines of the Clinical and Laboratory Standards Institute. RESULTS: E. raffinosus JARB-HU0741 contained a 3 248 808-bp chromosome and a 1 156 277-bp megaplasmid. cfr(B) was present in the Tn6218-like transposon, which was inserted into a gene encoding a PRD domain-containing protein present in the megaplasmid, but the isolate was susceptible to linezolid (MIC, 0.5 µg/mL). The Tn6218-like transposon was similar to the Tn6218 of Clostridioides difficile Ox3196 and the Tn6218-like transposon of Enterococcus faecium UW11733; however, three genes encoding a topoisomerase, an S-adenosylmethionine-dependent methyltransferase, and a TetR family transcriptional regulator were present in the previous Tn6218- or Tn6218-like transposon. CONCLUSION: This is the first report of the complete genome sequence of E. raffinosus carrying cfr(B). E. raffinosus carrying cfr(B) without linezolid resistance poses a threat, as it could serve as a reservoir for mobile linezolid resistance genes.

Complete genome sequence of optrA-carrying Enterococcus faecalis isolated from open pus in a Japanese patient.

OBJECTIVES: The occurrence of linezolid resistance in enterococci has recently increased. Here, we report the genomic characterization of Enterococcus faecalis strain JARB-HU0796-isolated from the open pus of a patient in Hiroshima, Japan-which shows nonsusceptibility to linezolid (MIC of 4 µg/mL). METHODS: JARB-HU0796 whole-genome sequencing was performed using short-read sequencing with Illumina Hiseq X Five and long-read sequencing using GridION. These reads were collected using the assembly pipeline Unicycler and annotated with DFAST. Antimicrobial resistance genes were detected using the Abricate and ResFinder databases, and the sequence type identified using PubMLST. The antimicrobial susceptibility of JARB-HU0796 was determined with the Eiken dry-plate QH02 system. RESULTS: The JARB-HU0796 complete genome contained a circular chromosome (2 722 585 bp) and two circular plasmids (85 996 bp and 58 872 bp). The chromosome harbours the optrA gene, which confers resistance to oxazolidinones and phenicols. JARB-HU0796 showed nonsusceptibility to linezolid and multidrug resistance to other antibiotics. MLST analysis identified JARB-HU0796 as ST476, similar to the optrA-positive E. faecalis ST476 isolates from swine (South Korea, 2020) and pet food (Switzerland, 2022). The optrA region of JARB-HU0796 is nearly identical to that of ST476 E. faecalis strain TZ2, isolated from humans (China, 2013). CONCLUSIONS: To the best of our knowledge, this is the first report of the complete genome sequence of E. faecalis ST476 carrying optrA on a chromosome isolated from a patient in Japan. The strain may have originated in animals, suggesting that the organisms acquired resistance to linezolid because the optrA gene may be closely spread between animals and humans.

Clinical characteristics of extrapulmonary nontuberculous mycobacteria infections in comparison with pulmonary infections: A single-center, retrospective study in Japan.

INTRODUCTION: The prevalence of nontuberculous mycobacteria (NTM) infections is increasing worldwide. Although NTM can affect extrapulmonary organs, studies on the clinical characteristics of extrapulmonary NTM are rare. METHODS: We retrospectively analyzed patients who were newly diagnosed with NTM infections at Hiroshima University Hospital between 2001 and 2021 to investigate species distribution, infected sites, and risk factors of extrapulmonary NTM compared to pulmonary NTM. RESULTS: Of the 261 NTM infections, 9.6% and 90.4% had extrapulmonary and pulmonary NTM, respectively. The mean ages of patients with extrapulmonary and pulmonary NTM were 53.4 and 69.3 years, 64.0% and 42.8% were male, 36.0% and 9.3% received corticosteroids, 20.0% and 0% had acquired immune deficiency syndrome (AIDS), and 56.0% and 16.1% had any immunosuppressive conditions, respectively. Younger age, corticosteroid use, and AIDS were associated with extrapulmonary NTM. In pulmonary NTM, Mycobacterium avium complex (MAC) accounted for 86.4% of NTM species, followed by M. abscessus complex (4.2%), whereas in extrapulmonary NTM, M. abscessus complex, MAC, M. chelonae, and M. fortuitum accounted for 36.0%, 28.0%, 12.0%, and 8.0%, respectively. Compared to pulmonary NTM, extrapulmonary NTM were significantly more likely to be rapid-growing mycobacteria (RGM) (56.0% vs. 5.5%). The most common sites of infection were the skin and soft tissues (44.0%), followed by the blood (20.0%), tenosynovium, and lymph nodes (12.0%). CONCLUSION: Younger age and immunosuppressive conditions are associated with extrapulmonary NTM, with a higher prevalence of RGM in extrapulmonary NTM than in pulmonary NTM. These results provide a better understanding of extrapulmonary NTM.

Prevalence of Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum detection in urine and respiratory tract samples in Hiroshima, Japan.

Background: Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum are commensal bacteria that are associated with colonization and infection of the urogenital tract. However, colonization of the respiratory tract by these microorganisms in adults has not been fully investigated. Methods: Urine and respiratory tract samples (sputum, tracheal aspirates, and bronchoalveolar lavage) of patients aged 20-80 years were analyzed to detect the presence of M. hominis, U. parvum, and U. urealyticum using a conventional PCR method. The samples were submitted to the microbiological clinical laboratory of Hiroshima University Hospital from December 1, 2021 to May 31, 2022. Results: In total, 334 urine and 238 respiratory tract samples were analyzed. The overall detection rates of M. hominis, U. parvum, and U. urealyticum were 2.9%, 1.7%, and 2.3% in male urine; 7.0%, 13.8%, and 1.9% in female urine; 2.2%, 0%, and 2.2% in male respiratory tract; and 0%, 2.0%, and 0% in female respiratory tract, respectively. In urine samples, the detection rates of M. hominis, U. parvum, and U. urealyticum were significantly higher (p < 0.001) for women (29/159; 18.2%) than for men (10/175; 5.7%); however, in respiratory tract samples, the detection rates were not significantly different (p = 0.70) between women (2/101; 2.0%) and men (5/137; 3.7%). Further, both the urine and respiratory samples of 83 patients were analyzed. Three male samples were positive for M. hominis or U. urealyticum, and M. hominis and U. urealyticum were matched in both the urine and respiratory tract samples: M. hominis (n = 1), U. urealyticum (n = 1), M. hominis + U. urealyticum (n = 1). Conclusion: M. hominis, U. parvum, and U. urealyticum were detected in the respiratory tract of not only the young patients, but also of patients aged 50-60 years. Further studies are required to understand the relationship of these microorganisms in urogenital and respiratory tract samples with extra-genital infections.

Clinical characteristics of Actinotignum schaalii bacteremia in a Japanese tertiary hospital.

We retrospectively analyzed seven patients with Actinotignum schaalii bacteremia in a tertiary hospital in Japan. Pyelonephritis was the most frequent source of bacteremia, followed by Fournier's gangrene and pyometra. All patients with pyelonephritis had underlying urological conditions, ureteral stents, nephrostomy, ureteral stones, or ureterocele.

Vertebral osteomyelitis caused by Mycobacteroides abscessus subsp. abscessus resulting in spinal cord injury due to vertebral body fractures.

Nontuberculous mycobacteria (NTM) rarely cause vertebral osteomyelitis; however, the clinical characteristics of vertebral osteomyelitis caused by NTM are poorly understood due to its rarity. A 74-year-old man with lung cancer was treated with prednisolone for immune checkpoint inhibitor-associated immune-related adverse events. He had been experiencing mild back pain without febrile episodes for five months, and was admitted to the hospital for worsening back pain and progressive paraplegia. Magnetic resonance imaging showed spinal cord compression at T4-5 due to fractures of the T5 and T7 vertebral bodies. The culture of a sample of pus from the T7 vertebral body obtained at the time of spinal fusion surgery yielded the Mycobacteroides abscessus (M. abscessus) complex. The patient was diagnosed with vertebral osteomyelitis caused by M. abscessus complex and treated with clarithromycin, amikacin, and imipenem; clarithromycin was later replaced by sitafloxacin because of inducible macrolide resistance. However, his neurologic deficits were irreversible, and he died due to a deteriorating general condition. The strain was identified up to subspecies level as M. abscessus subsp. abscessus by hsp65 and rpoB sequencing and nucleic acid chromatography. Although vertebral osteomyelitis due to NTM is rare, delayed diagnosis can lead to serious complications or poor outcomes. A prolonged clinical course, less frequent fever, vertebral destruction or spinal deformity, neurological deficits, or immunosuppressed conditions might be suggestive of NTM vertebral osteomyelitis.

Laboratory identification and clinical characteristics of Streptococcus bovis/Streptococcus equinus complex bacteremia: a retrospective, multicenter study in Hiroshima, Japan.

BACKGROUND: Bacteremia due to the Streptococcus bovis/Streptococcus equinus complex (SBSEC) is associated with specific diseases, such as colorectal cancer and infective endocarditis. This study aimed to evaluate the clinical characteristics of SBSEC bacteremia and the accuracy of identification of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and phenotypic identification systems for SBSEC isolates. METHODS: We analyzed patients with SBSEC bacteremia retrospectively between 2012 and 2019 at three hospitals in Japan. We re-identified each SBSEC isolate using sequencing superoxide dismutase (sodA) analysis, MALDI-TOF MS using the MALDI Biotyper, and phenotypic identification using the VITEK2. RESULTS: During the study period, 39 patients with SBSEC bacteremia were identified. S. gallolyticus subsp. pasteurianus (SGSP, n = 29), S. gallolyticus subsp. gallolyticus (SGSG, n = 5), S. lutetiensis (SL, n = 4), and S. infantarius subsp. infantarius (n = 1) were identified using sodA sequencing analysis. Primary bacteremia (36%) was the most common cause of bacteremia, followed by infective endocarditis (26%) and biliary tract infections (23%). Colorectal cancer was associated significantly with SGSG bacteremia, while the sources of bacteremia were similar in each SBSEC subspecies. The MALDI Biotyper was significantly more accurate in identifying the SBSEC isolates at the subspecies level compared to the VITEK2 (92% vs. 67%, P = 0.010). In contrast, there were no significant differences in the rates of correct identification of the SBSEC isolates at the species level between the MALDI Biotyper and the VITEK2 (100% vs. 87%, P = 0.055). CONCLUSIONS: Bacteremia with SGSG was associated with colorectal cancer, and the sources of bacteremia were similar in each SBSEC subspecies. The MALDI-TOF MS was significantly more accurate in identifying SBSEC isolates at the subspecies level than the phenotypic identification systems. The accurate identification of SBSEC isolates using the MALDI-TOF MS and phenotypic identification systems was sufficient at the species level, but it was insufficient at the subspecies level. Therefore, it may be reasonable for clinicians to perform echocardiographies and colonoscopies in all patients with SBSEC bacteremia.

Postoperative mediastinitis after cardiac surgery caused by Mycoplasma hominis: a case report.

BACKGROUND: Mycoplasma hominis is a human commensal bacterium of the urogenital tract, and extragenital infection caused by M. hominis has rarely been reported. The identification of M. hominis is challenging, and surgeons are generally not aware that this bacteria can cause postoperative infection. Here, we report a rare case of postoperative mediastinitis caused by M. hominis after cardiac surgery in an immunocompetent patient. CASE PRESENTATION: A 54-year-old man presented with pain and purulent discharge from the wound after aortic valve replacement and patent foramen ovale closure. However, Gram staining and culture of bacteria from the purulent discharge was negative, and empiric sulbactam/ampicillin therapy was not effective. This patient developed mediastinitis and rupture of a pseudoaneurysm of the ascending aorta caused by mediastinitis, and re-operation was performed. Then, postoperative mediastinitis caused by M. hominis or Ureaplasma species was suspected and bacterial cultures targeting these pathogens were performed. M. hominis was identified from abscess and tissue obtained from the surgical site and urine. A final diagnosis of postoperative mediastinitis caused by M. hominis was determined. The patient was initially treated with levofloxacin and then with minocycline for 3 weeks. The patient's clinical condition improved; the patient was transferred to another hospital. CONCLUSION: The role of M. hominis as a cause of postoperative infection might be underestimated in cardiac surgery. M. hominis should be considered when culture-negative purulent discharge is observed or there is no response to standard empiric treatment of postoperative infections.

The effect of pulsed-xenon ultraviolet disinfection on surfaces contaminated with vancomycin-resistant Enterococci in a Japanese hospital.

INTRODUCTION: Recently, a worldwide outbreak of vancomycin-resistant Enterococci (VRE) was reported. However, due to the low incidence of VRE infection and colonization, VRE contamination of hospital environments has not been fully investigated in Japan. METHODS: Surfaces were swabbed, before and after manual cleaning and after pulsed xenon ultraviolet (PX-UV) disinfection, in five patient rooms that had been occupied by patients colonized with VRE. Difference in the number of VRE-positive samples and VRE colony forming units (CFUs), before and after disinfection, for each cleaning method was estimated. RESULTS: We detected VRE contamination in 22/60 (37%) and 14/60 (23%) samples collected before and after manual cleaning, respectively. In contrast, VRE contamination was not detected in the samples collected after PX-UV disinfection. In addition, 3/5 (60%) spray nozzles of electric warm-water bidet toilet seats were found to be contaminated with VRE before terminal cleaning. Manual cleaning caused a significant decrease in the number of VRE CFUs compared with that before cleaning (P = 0.031). PX-UV disinfection also caused a significant decrease in the number of VRE CFUs compared to that of manual cleaning (P < 0.001). CONCLUSION: We identified hot spots of severe contamination, such as private bathrooms in patient rooms and areas around the bed of patients using diapers and required assistance. VRE contamination persisted even after terminal disinfection; PX-UV disinfection in addition to terminal disinfection was effective at eliminating VRE contamination. These results can be useful in controlling the spread of VRE infections in Japanese hospitals.

Pilot study on the decontamination efficacy of an installed 222-nm ultraviolet disinfection device (Care222™), with a motion sensor, in a shared bathroom.

BACKGROUND: Toilet surfaces are contaminated with pathogens, and they may be a vector for disease transmission. In this study, we investigated the efficacy of an automated 222-nm ultraviolet C (UVC) disinfection device "Care222™," with a motion sensor, for removing bacterial contamination in a shared bathroom. METHODS: Two automated UVC devices, deactivated by motion sensors, were mounted on the ceiling of two bathrooms; the emission window of the UVC device was covered in the non-treated bathroom. After irradiation, samples were collected from five surfaces at four time points/day for 5 days, and colony-forming units (CFUs) of aerobic bacteria (AB) were determined. The irradiation time was also measured. RESULTS: UVC source deactivation time did not significantly differ between the bathrooms. There was a significant difference in the total AB CFUs between the treated and non-treated bathrooms. In the treated bathroom, the CFUs of AB of the toilet seat, control panel of the electric toilet seat, and top of the toilet paper holder were significantly lower than those of the control. The CFUs of AB at 9:00, 15:00, and 18:00 h in the treated bathroom were significantly lower than those of the control. CONCLUSIONS: The automated 222-nm UVC disinfection device with a motion sensor significantly reduced AB surface contamination of a shared bathroom.

Clinical and microbiological characteristics of Eggerthella lenta bacteremia at a Japanese tertiary hospital.

Eggerthella lenta is an important cause of anaerobic bloodstream infections and is associated with high mortality. However, there are few reports of E. lenta infection in Japan. This study aimed to evaluate the clinical and microbiological characteristics of bacteremia caused by E. lenta in Hiroshima, Japan. We retrospectively analyzed E. lenta bacteremia patients at the Hiroshima University Hospital between January 2012 and December 2020. During the study period, 14 patients with E. lenta bacteremia were identified. All E. lenta isolates were cultured in anaerobic bottles, and the median time to blood culture positivity was 52.9 h. In most cases (85.6%), the source of E. lenta bacteremia was associated with intra-abdominal infections, and colon perforation was the most frequent source of E. lenta bacteremia (42.9%, n = 6). Antimicrobial susceptibility testing showed high minimal inhibitory concentrations (MIC) of piperacillin-tazobactam (TZP) and 100% susceptibility to ampicillin-sulbactam, carbapenems, and metronidazole. This study demonstrates that E. lenta bacteremia is associated with intra-abdominal infections, particularly colon perforation, and a high MIC of TZP. When gram-positive anaerobes are detected in the blood cultures of patients with severe intra-abdominal infections, clinicians should suspect E. lenta, and it may be better to change antimicrobial agents from TZP.

Efficacy of pulsed xenon ultraviolet disinfection of multidrug-resistant bacteria and Clostridioides difficile spores.

BACKGROUND: Contamination of healthcare environments by multidrug-resistant organisms (MDRO) and Clostridioides difficile is a risk for healthcare-associated infections. The efficacy of pulsed xenon ultraviolet (PX-UV) disinfection in healthcare environments has been described previously. However, there are few reports about PX-UV disinfection in Japan. The aim of this study was to investigate in vitro the efficacy of PX-UV disinfection of MDRO and C. difficile spores commonly isolated in Japanese hospitals. METHODS: We investigated reductions in microbial counts after exposure to PX-UV of the following clinically-isolated organisms on seeding agar plates: methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecium, carbapenemase-producing Klebsiella pneumoniae, extended spectrum ß-lactamase-producing Escherichia coli, multidrug resistant Acinetobacter baumannii, and C. difficile spores. We also visually assessed the attenuation of disinfection by shielding of MRSA and carbapenemase-producing K. pneumoniae from PX-UV exposure. RESULTS: PX-UV disinfection for 5 min induced >5-log colony-forming units (CFU)/cm2 growth inhibition of all the MDRO. PX-UV disinfection for 15 min induced >3-log CFU/cm2 growth inhibition of C. difficile spores. Where a plate was shielded from PX-UV exposure the bacteria showed confluent growth, but no colonies were observed on unshielded (exposed) parts of the plates. CONCLUSION: This study shows the efficacy of PX-UV disinfection against clinical MDROs. C. difficile spores were more resistant to PX-UV disinfection than vegetative bacteria. Further evaluation of the efficacy of PX-UV disinfection in reducing the contamination of real-world surfaces and the incidence of healthcare-associated infections is needed.

Performance of laboratory tests for detection for Clostridioides difficile: A multicenter prospective study in Japan.

BACKGROUND: The optimal and practical laboratory diagnostic approach for detection of Clostridioides difficile to aid in the diagnosis of C. difficile infection (CDI) is controversial. A two-step algorithm with initial detection of glutamate dehydrogenase (GDH) or nucleic acid amplification test (NAAT) alone are recommended as a predominant method for C. difficile detection in developed countries. The aim of this study was to compare the performance of enzyme immunoassays (EIA) detecting toxins A and B, NAAT detecting the toxin B gene, and GDH compared to toxigenic culture (TC) for C. difficile as the gold standard, in patients prospectively and actively assessed with clinically significant diarrhea in 12 medical facilities in Japan. METHODS: A total of 650 stool specimens were collected from 566 patients with at least three diarrheal bowel movements (Bristol stool grade 6-7) in the preceding 24 h. EIA and GDH were performed at each hospital, and NAAT and toxigenic C. difficile culture with enriched media were performed at the National Institute of Infectious Diseases. All C. difficile isolates recovered were analyzed by PCR-ribotyping. RESULTS: Compared to TC, the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of EIA were 41%, 96%, 75% and 84%, respectively, and for NAAT were 74%, 98%, 91%, and 92%, respectively. In 439 specimens tested with GDH, the sensitivity, specificity, PPV, and NPV were 73%, 87%, 65%, and 91%, and for an algorithm (GDH plus toxin EIA, arbitrated by NAAT) were 71%, 96%, 85%, and 91%, respectively. Among 157 isolates recovered, 75% of isolates corresponded to one of PCR-ribotypes (RTs) 002, 014, 018/018", and 369; RT027 was not isolated. No clear differences in the sensitivities of any of EIA, NAAT and GDH for four predominant RTs were found. CONCLUSION: The analytical sensitivities of NAAT and GDH-algorithm to detect toxigenic C. difficile in this study were lower than most previous reports. This study also found low PPV of EIAs. The optimal method to detect C. difficile or its toxins to assist in the diagnosis of CDI needs further investigation.

Clostridioides (Clostridium) difficile infection burden in Japan: A multicenter prospective study.

Clostridioides (Clostridium) difficile is the leading cause of healthcare-associated infectious diarrhea in the developed world. Retrospective studies have shown a lower incidence of C. difficile infection (CDI) in Japan than in Europe or North America. Prospective studies are needed to determine if this is due lack of testing for C. difficile or a true difference in CDI epidemiology. A prospective cohort study of CDI was conducted from May 2014 to May 2015 at 12 medical facilities (20 wards) in Japan. Patients with at least three diarrheal bowel movements (Bristol stool grade 6-7) in the preceding 24 h were enrolled. CDI was defined by positive result on enzyme immunoassay for toxins A/B, nucleic acid amplification test for the toxin B gene or toxigenic culture. C. difficile isolates were subjected to PCR-ribotyping (RT), slpA-sequence typing (slpA-ST), and antimicrobial susceptibility testing. The overall incidence of CDI was 7.4/10,000 patient-days (PD). The incidence was highest in the five ICU wards (22.2 CDI/10,000 PD; range: 13.9-75.5/10,000 PD). The testing frequency and CDI incidence rate were highly correlated (R2 = 0.91). Of the 146 isolates, RT018/018″ was dominant (29%), followed by types 014 (23%), 002 (12%), and 369 (11%). Among the 15 non-ICU wards, two had high CDI incidence rates (13.0 and 15.9 CDI/10,000 PD), with clusters of RT018/slpA-ST smz-02 and 018"/smz-01, respectively. Three non-RT027 or 078 binary toxin-positive isolates were found. All RT018/018" isolates were resistant to moxifloxacin, gatifloxacin, clindamycin, and erythromycin. This study identified a higher CDI incidence in Japanese hospitals than previously reported by actively identifying and testing patients with clinically significant diarrhea. This suggests numerous patients with CDI are being overlooked due to inadequate diagnostic testing in Japan.