Performance of the Assays for Hepatitis B Virus DNA using the µTASWako g1 in Comparison with the COBAS TaqMan Test.

BACKGROUND: The quantification of hepatitis B virus (HBV) DNA is used to monitor antiviral treatment for HBV infection. Recently, an HBV-DNA quantification reagent and assay protocol have been developed for the µTASWako g1 (Fujifilm Wako Pure Chemical Corporation, Osaka, Japan), a fully automated genetic analyzer that uses PCR-capillary electrophoresis. We evaluated the performance of the newly developed µTASWako HBV-DNA assay using standard and clinical samples. METHODS: The performance of µTASWako HBV-DNA was evaluated using 3rd World Health Organization International Standard for HBV. Thereafter, we evaluated the correlation between the serum HBV DNA concentrations obtained using the µTASWako HBV-DNA and the Roche Cobas AmpliPrep/COBAS TaqMan HBV test, version 2.0 (CAP/CTM HBV test v.2.0) using 190 serum samples from possible HBV carriers. RESULTS: The limit of detection of the µTASWako HBV-DNA was 7.1 IU/mL and that of the CAP/CTM HBV test v.2.0 was 14.6 IU/mL. Seventy-six of the 190 samples yielded values between 1.3 - 7.8 log IU/mL from both assays. The correlation between the results of the assays for these samples was good, with a Deming regression equation of y = 0.929x + 0.041, 95% confidence intervals (CI) for the slope and intercept of 0.892 - 1.12 and -0.474 - 0.110, respectively, and a correlation coefficient r = 0.924. In the low concentration range of 1.3 - 4.0 log IU/mL (n = 64), the Deming regression equation was y = 0.893x + 0.126, and the 95% CIs for the slope and intercept were 0.915 - 1.342 and -0.930 - 0.025, respectively, and r = 0.809. There was also a close correlation for HBV DNA genotype C (n = 41), with a Deming regression equation of y = 0.975x - 0.048, 95% CIs of the slope and intercept of 0.872 - 1.183 and -0.591 - 0.188, respectively, and r = 0.950. The correlations of the four HBV DNA level categories (not detected, < LLOQ (< 1.3 log IU/mL), 1.3 - 3.2 log IU/mL, and ≥ 3.3 log IU/mL) between the two assays for the 190 samples was also compared, and the overall concordance rate was 81.6% (155/190), with a κ statistic of 0.73 (standard error, 0.040; 95% CI, 0.647 - 0.803). CONCLUSIONS: The µTASWako HBV-DNA has a performance comparable with that of the CAP/CTM HBV test, v.2.0. Thus, µTASWako HBV-DNA is useful for the monitoring of HBV infection.

Antimicrobial Susceptibility to 27 Drugs and the Molecular Mechanisms of Macrolide, Tetracycline, and Quinolone Resistance in Gemella sp.

Gemella is a catalase-negative, facultative anaerobic, Gram-positive coccus that is commensal in humans but can become opportunistic and cause severe infectious diseases, such as infective endocarditis. Few studies have tested the antimicrobial susceptibility of Gemella. We tested its antimicrobial susceptibility to 27 drugs and defined the resistant genes using PCR in 58 Gemella strains, including 52 clinical isolates and six type strains. The type strains and clinical isolates included 22 G. morbillorum, 18 G. haemolysans (GH) group (genetically indistinguishable from G. haemolysans and G. parahaemolysans), 13 G. taiwanensis, three G. sanguinis, and two G. bergeri. No strain was resistant to beta-lactams and vancomycin. In total, 6/22 (27.3%) G. morbillorum strains were erythromycin- and clindamycin-resistant ermB-positive, whereas 4/18 (22.2%) in the GH group, 7/13 (53.8%) G. taiwanensis, and 1/3 (33.3%) of the G. sanguinis strains were erythromycin-non-susceptible mefE- or mefA-positive and clindamycin-susceptible. The MIC90 of minocycline and the ratios of tetM-positive strains varied across the different species-G. morbillorum: 2 µg/mL and 27.3% (6/22); GH group: 8 µg/mL and 27.8% (5/18); G. taiwanensis: 8 µg/mL and 46.2% (6/13), respectively. Levofloxacin resistance was significantly higher in G. taiwanensis (9/13 69.2%) than in G. morbillorum (2/22 9.1%). Levofloxacin resistance was associated with a substitution at serine 83 for leucine, phenylalanine, or tyrosine in GyrA. The mechanisms of resistance to erythromycin and clindamycin differed across Gemella species. In addition, the rate of susceptibility to levofloxacin differed across Gemella sp., and the quinolone resistance mechanism was caused by mutations in GyrA alone.

First report of New Delhi metallo-ß-lactamase-1-producing Acinetobacter soli in Japan.

New Delhi metallo-ß-lactamase (NDM)-producing gram-negative rods, including Acinetobacter species, are a global problem but have rarely been isolated in Japan. To our knowledge, this is the first study to isolate an NDM-1-producing Acinetobacter soli strain, KUH106, in Japan. We analyzed this strain using next-generation sequencing to examine the plasmid carrying NDM-1. This plasmid, named pKUH106_NDM1, is 41,135 bp in length and contains genetic contexts with the structure ISAba14-aph(3')-VI-ISAba125-blaNDM-1ble-MBL. Comparative analysis of the plasmid revealed that it resembled the plasmids of Acinetobacter detected in various countries, such as the A. soli isolate from Taiwan and the Acinetobacter baumannii isolate from a healthcare facility in Osaka Prefecture, Japan. These results suggest that blaNDM-1 may spread via this plasmid in Acinetobacter species. This phenomenon needs to be confirmed through the genetic analysis of A. baumannii and other carbapenem-resistant Acinetobacter species. In particular, blaNDM-1 and other resistance genes must be investigated, and the spread of these genes in the community must be cautioned.

Purpura fulminans with Lemierre's syndrome caused by Gemella bergeri and Eikenella corrodens: a case report.

BACKGROUND: Gemella bergeri is one of the nine species of the genus Gemella and is relatively difficult to identify. We herein describe the first case of septic shock due to a Gemella bergeri coinfection with Eikenella corrodens. CASE PRESENTATION: A 44-year-old Asian man with a medical history of IgG4-related ophthalmic disease who was prescribed corticosteroids (prednisolone) presented to our hospital with dyspnea. On arrival, he was in shock, and a purpuric eruption was noted on both legs. Contrast enhanced computed tomography showed fluid retention at the right maxillary sinus, left lung ground glass opacity, and bilateral lung irregular opacities without cavitation. Owing to suspected septic shock, fluid resuscitation and a high dose of vasopressors were started. In addition, meropenem, clindamycin, and vancomycin were administered. Repeat computed tomography confirmed left internal jugular and vertebral vein thrombosis. Following this, the patient was diagnosed with Lemierre's syndrome. Furthermore, he went into shock again on day 6 of hospitalization. Additional soft tissue infections were suspected; therefore, bilateral below the knee amputations were performed for source control. Cultures of the exudates from skin lesions and histopathological samples did not identify any pathogens, and histopathological findings showed arterial thrombosis; therefore it was concluded that the second time shock was associated with purpura fulminans. Following this, his general status improved. He was transferred to another hospital for rehabilitation. The blood culture isolates were identified as Gemella bergeri and Eikenella corrodens. Gemella bergeri was identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry and confirmed by 16S rRNA gene sequencing later. The primary focus of the infection was thought to be in the right maxillary sinus, because the resolution of the fluid retention was confirmed by repeat computed tomography. CONCLUSIONS: Gemella bergeri can be the causative pathogen of septic shock. If this pathogen cannot be identified manually or through commercial phenotypic methods, 16S rRNA gene sequencing should be considered.

[Efforts of medical technologists for cancer patients--a questionnaire survey following a lecture meeting: "cancer treatment and clinical examinations" for cancer patients and their families].

As interprofessional work in cancer treatment becomes increasingly important, medical technologists are required to play active roles as part of the team. The cancer center of our hospital organized a lecture meeting, "The 6th Lecture Meeting: Living Together," for cancer patients and their families, and a medical technologist presented a lecture entitled: "Cancer treatment and clinical examinations." According to the results of a questionnaire survey conducted following the meeting, most participants were able to understand the lecture and were satisfied with it. Based on the opinions expressed by meeting participants and the questionnaire results, medical technologists initiate the following services and activities: 1. They explain the results of white blood cell and neutrophil counts of patients on chemotherapy and/or radiation therapy; and 2. provide medical examinations and consultation at outpatient chemotherapy centers. We believe that these efforts will help improve cancer treatment and further contribute to interprofessional health care.