Data concerning the human microbiota composition during Clostridioides (Clostridium) difficile infection (CDI) using next-generation sequencing are still limited. We aimed to confirm key features indicating tcdB positive patients and compare the microbiota composition between subgroups based on toxin gene load ( tcdB gene) and presence of significant diarrhea. Ninety-nine fecal samples from 79 tcdB positive patients and 20 controls were analyzed using 16S rRNA gene sequencing. Chao1 index for alpha diversity were calculated and principal coordinate analysis was performed for beta diversity using Quantitative Insights into Microbial Ecology (QIIME) pipeline. The mean relative abundance in each group was compared at phylum, family, and genus levels. There were significant alterations in alpha and beta diversity in tcdB positive patients (both colonizer and CDI) compared with those in the control. The mean Chao1 index of tcdB positive patients was significantly lower than the control group ( P <0.001), whereas there was no significant difference between tcdB groups and between colonizer and CDI. There were significant differences in microbiota compositions between tcdB positive patients and the control at phylum, family, and genus levels. Several genera such as Phascolarctobacterium , Lachnospira , Butyricimonas , Catenibacterium , Paraprevotella , Odoribacter , and Anaerostipes were not detected in most CDI cases. We identified several changes in the microbiota of CDI that could be further evaluated as predictive markers. Microbiota differences between clinical subgroups of CDI need to be further studied in larger controlled studies.

In this study, we aimed to evaluate the composition of the intestinal microbiota and level of fecal calprotectin in Clostridioides difficile-colonized patients. We included 102 C. difficile non-colonized (group I), 93 C. difficile colonized subjects (group II), and 89 diarrhea patients with C. difficile (group III). Chao1 index for alpha diversity and principal coordinate analysis was performed for beta diversity using QIIME. The mean relative abundance in each group was compared at the phylum and genus levels. Fecal calprotectin was measured using EliA calprotectin (Thermo Fisher Scientific). Group II showed significantly lower levels of Sutterella, Blautia, Ruminococcus, Faecalibacterium, Bilophila, and Ruminococcaceae and higher levels of Enterobacteriaceae compared to group I (p = 0.012, 0.003, 0.002, 0.001, 0.027, 0.022, and 0.036, respectively). Toxigenic C. difficile colonized subjects showed significantly lower levels of Prevotella, Phascolarctobacterium, Succinivibrio, Blautia, and higher levels of Bacteroides. The level of fecal calprotectin in group III was significantly higher than those in group I and group II (p < 0.001 for both). These data could be valuable in understanding C. difficile colonization process and the microbiota and inflammatory markers could be further studied to differentiate colonization from CDI.

Background: Although plasma free hemoglobin (fHb) test is important for assessing intravascular hemolysis, it is still dependent on the gold standard Harboe method using manual and labor-intensive spectrometric measurements at the wavelength of 380-415-450 nm. We established an automated fHb assay using a routine chemistry autoanalyzer that can be tuned to a wavelength of 380-416-450 nm. Methods:The linearity, precision, accuracy, correlation, and sample carryover of fHb measurement using TBA200FRneo method and manual Harboe method were evaluated, respectively. fHb values measured by manual Harboe method were compared with those measured by our new automated TBA200FRneo method.Results: fHb measurements were linear in the range of 0.05~38.75 µmol/L by TBA200FRneo and 0.05~9.69 µmol/L by manual Harboe method. Imprecision analysis (%CV) revealed 0.9~2.8% for TBA200FRneo method and 5.3~13.6% for the manual Harboe method. Comparison analysis showed 0.9986 of correlation coefficient (T BA200FRneo = 0.970 × Harboe + 0.12). In analytical accuracy analysis, the manual Harboe method revealed about 4 times higher average total error % (12.2%) than the TBA200FRneo automated method (2.8%). The sample carryover was −0.0016% in TBA200FRneo method and 0.0038% in Harboe method. Conclusions:In the measurement of fHb, the automated TBA200FRneo method showed better performance than the conventional Harboe method. It is expected that the automated fHb assay using the routine chemistry analyzer can replace the gold standard Harboe method which is labor-intensive and need an independent spectrophotometry equipment.

No abstract

Under the Korean Association of External Quality Assessment Service, autoimmune proficiency testings (PT) for six test items were performed in 2018-2019 for laboratory quality improvement. We conducted two trials per year and sent three PT materials for anti-nuclear antibody (ANA) testing and two PT materials for anti-mitochondrial antibody (AMA), anti-smooth muscle antibody (SMA), anti-thyroglobulin antibody (anti-Tg), anti-thyroperoxidase antibody (anti-TPO), and anti-double stranded DNA antibody (anti-dsDNA) testing in each trial. The analysis was conducted based on the information and results of each test item entered by the laboratory. The report comprised of a common report that showed the characteristics of all the participating laboratories and a laboratory-specific report that showed the assessment data of individual laboratories. The intended response rates for ANA, AMA, SMA, and anti-dsDNA qualitative tests were over 97.5%, 88.2%, 85.0%, and 90.4%, respectively. The coefficient of variations for anti-Tg and anti-TPO was 10.4%-70.1% and 16.6%-21.0%, respectively. The number of participating laboratories in 2019 was more than that in 2018. We believe this statistical report will be useful to interpret external PT results and set up autoimmune assays at each laboratory.