Biological products, such as vaccines, blood products, antitoxins, and antivenoms, are released into the market following a lot release conducted by National Regulatory Authorities or National Control Laboratories, even if their manufacturing and marketing have been authorized. Independent lot release by regulatory authorities is not a procedure unique to Japan, but is performed worldwide. Previously, Japan carried out lot release mainly by laboratory tests, and the manufacturers’ in-house test records were used as a reference, not involved in the decision of lot release. Conversely, the international standard procedure promoted by the World Health Organization (WHO) includes a document review of the manufacturers’ summary protocols, and laboratory tests are listed as an optional procedure. To harmonize with the WHO recommended international method, Japan modified the procedure and introduced a document review in addition to laboratory tests for vaccines in 2012. Since then, substantial knowledge regarding vaccine quality has been obtained during the process of summary protocol reviewing. Here, we outline the current status of the lot release procedure in Japan. We shed light on its history and show recent research based on the knowledge obtained from the protocol review to improve efficiency of laboratory testing and international harmonization.
To assess the clinical characteristics of the rare Kluyvera ascorbata infection, we reviewed the medical records of patients from whom K. ascorbata was isolated from 2010 to 2016, and conducted a systematic review of the English and Spanish literature in PubMed for reports of K. ascorbata infection in humans from 1971 to 2018. A total of 43 cases (24 adults and 19 children) were enrolled: 3 at our hospital and 40 from the literature review. The urinary tract was the most common site of infection (44.2%, 19/43), followed by the bloodstream (27.9%, 12/43). There was no significant difference in the frequency of urinary tract infections (50% vs 36.8%; P = 0.388) and bloodstream infections (25% vs 31.6%; P = 0.633) in adults and children. Seventeen (60.7%, present in 28 of 43 cases) had nosocomial or healthcare-associated infections: 72.7% among children and 60% among adults. Superinfection developed in 20% (6 in 30 cases). The overall mortality was 12.1%. The antimicrobial agents mainly used in these 43 cases were third-generation cephalosporin, cefepime, piperacillin-tazobactam, ciprofloxacin, amikacin, and carbapenem. Most strains were resistant to ampicillin and first- and second-generation cephalosporins. K. ascorbata is a rare but significant clinical pathogen in adults and children.
In Japan, 92.6% of antibiotics consumed are oral agents; most of these are for outpatients. A significant proportion is known to be dispensed for children; however, the specific pattern of antibiotic prescription in accordance with clinical specialty is still unclear. The aim of our study was to identify the key targets for the optimization of oral antibiotic use in children. We analyzed data on oral antimicrobial prescription patterns for children ＜ 16 years old of age in 3 urban districts by using a national database in Japan. Oral prescriptions were categorized according to their class, spectrum, clinical specialty, and type of clinical setting. The antibiotic spectrum was categorized as narrow, broad, or ultra-broad. In total, 132,869,332 antibiotic prescriptions were collected for analysis. The proportions of narrow-spectrum, broad-spectrum, and ultra-broad-spectrum antibiotics were 10.9%, 73.7%, and 15.4% in primary care clinics and 23.4%, 71.1%, and 5.4% in hospitals, respectively. Prescriptions from pediatricians and otolaryngologists in primary care clinics were predominant in the 3 studied areas. Third-generation cephalosporins, quinolones, penems, and carbapenems were prescribed mostly by pediatricians and otolaryngologists. Ultra-broad spectrum antibiotics used in primary care clinics and antibiotics particular to each specialty were identified as key targets for the optimization of oral antibiotic use for pediatric outpatients.
There is a paucity of data regarding the differentiating characteristics of patients with laboratory-confirmed and those negative for Middle East respiratory syndrome coronavirus (MERS-CoV) in South Korea. This hospital-based retrospective study compared MERS-CoV-positive and MERS-CoV-negative patients. A total of seven positive patients and 55 negative patients with a median age of 43 years (P = 0.845) were included. No statistical differences were observed with respect to their sex and the presence of comorbidities. At the time of admission, headache (28.6% vs. 3.6%; odds ratio [OR], 10.60; 95% confidence interval [CI], 1.22–92.27), myalgia (57.1% vs. 9.1%; OR, 13.33; 95% CI, 2.30–77.24), and diarrhea (57.1% vs. 14.5%; OR, 7.83; 95% CI, 1.47–41.79) were common among MERS-CoV-positive patients. MERS-CoV-positive patients were more likely to have a low platelet count (164 ± 76.57 vs. 240 ± 79.87) and eosinophil (0.27 ± 0.43 vs. 2.13 ± 2.01; P = 0.003). Chest radiography with diffuse bronchopneumonia was more frequent in MERS-CoV-positive patients than in negative patients (100% vs. 62.5%; P = 0.491). The symptoms of headache, myalgia, and diarrhea, as well as laboratory characteristics, including low platelet counts and eosinophil, and chest X-ray showing diffuse bronchopneumonia might enhance the ability to detect patients in South Korea infected with MERS-CoV.
We evaluated the prevalence of Anaplasma infection in 332 dogs from Ibaraki, Japan, using serological and molecular methods. An immunofluorescence antibody assay against Anaplasma phagocytophilum indicated that 7 of the 328 serum samples tested (2.1%) were positive for A. phagocytophilum. Screening by polymerase chain reaction (PCR) analysis demonstrated that 8 of the 331 peripheral blood samples tested (2.4%) were positive for Anaplasmataceae. Phylogenetic analysis of the partial 16S rRNA sequence of the PCR amplicons revealed that 6 sequences were most similar to the 16S rRNA sequence of a Wolbachia sp., and the remaining 2 to A. bovis. Further analysis by A. phagocytophilum-specific nested PCR demonstrated that 1 dog infected with A. bovis was also positive for A. phagocytophilum. This is the first study to report the dual infection of a dog in Japan with A. bovis and A. phagocytophilum.
We aimed to assess the 24-week virological and immunological success of the treatment of treatment-naive and treatment-experienced patients included in the Action against HIV in Istanbul (ACTHIV-IST) database. The ACTHIV-IST database was screened retrospectively from January 2012 to January 2014. The data for these patients such as age, sex, treatment-naive or treatment-experienced status, date of diagnosis, date of commencing antiretroviral therapy, antiretroviral therapy regimen, CD4+ cell count, and viral load before and after therapy were analyzed. In the 24th week of antiretroviral therapy, there were 40 (17.9%) and 29 (14.1%) virological and immunological failures, respectively. Virological failure (VF) was associated with a baseline viral load > 100,000 copies (p = 0.004). A CD4+ cell count lower than 200 cells/μl was not found to be associated with VF (p = 0.843). Immunological failure was substantially rare in patients with a baseline CD4+ cell count > 200 cells/μl (p = 0.005). Although an HIV-RNA ≤ 100,000 copies/ml was protective against VF in the 24th week, in individuals with an HIV-RNA > 100,000 copies/ml, VF was 3.2 times more likely to occur. Baseline VF was the most predictive parameter to estimate 24th week virological success and VF. VF is an important prognostic parameter resulting in CD4+ cell depletion, AIDS-related events, and increased mortality.
The increase in antimicrobial-resistant (AMR) bacteria caused by antimicrobial usage is a public health problem. We investigated the proportion of cephalexin (LEX)-resistant bacteria in fresh feces obtained from antimicrobial-free broilers in three flocks at ＜15, 15–40, and＞ 40 days old. DHL agar plates containing 25 μg/mL LEX (DHL-L) showed LEX-resistant bacteria in all flocks at ＜15 days old and in one flock at ＞ 40 days old. The bacterial counts on DHL and DHL-L were 105–108 colony forming units (CFU)/g feces and ＜102–105 CFU/g feces, respectively. We also assessed the proportion of AMR bacteria in feces collected at 5, 12, 19, 26, 33, and 40 days old from two flocks treated with amoxicillin at 5–7 days old and co-trimoxazole at 24–26 days old. The proportion of ampicillin (AMP)-resistant bacteria was elevated at 12 and 26–33 days old on DHL containing 50 μg/mL AMP, while no increase in LEX-resistant bacteria was observed on DHL-L. All isolates tested exhibited AMP resistance at 12 days old, while most exhibited resistance to both AMP and trimethoprim/sulfamethoxazole at 26–33 days old. Our results suggest that antimicrobial administration influenced the selection of AMR bacteria with cross- and coresistance.
In Turkey, the Measles Elimination Program has been implemented since 2002. The aim of this study was to evaluate the measles-specific antibody levels of mothers admitted to a hospital for birth and their infants, to determine the factors influencing the antibody levels of both, and to evaluate the transplacental transport ratio. We selected healthy women who came to the hospital for birth and their healthy newborns. We collected blood samples from 1,547 mothers and 1,529 infants. The protective prevalence of measles antibody levels of mothers was 80% (95% confidence interval [CI]: 78–82%) and that of newborns was 85% (95% CI: 83–86%). The antibody levels of mothers and newborns were positively linearly correlated (R: 0.922, p < 0.001) and were associated with parity (p < 0.001). The ratio of neonatal to maternal antibody levels increased with gestational age. The protective levels were 1.6 times higher (95% CI: 1.1–2.4) in mothers ≥ 32 years of age and 2.1 times higher (95% CI: 1.4–3.3) in naturally immune mothers. Two factors affecting the antibody levels of newborns were the mothers’ antibody levels and their immunization status. The antibody level of mother was the most significant factor that influenced the infant’s antibody level. Vaccination of women before pregnancy could enhance passive antibody protection by increasing the level of transplacental transmission.
A capnophilic Gram-negative rod-shaped bacterium was recovered from the urine of an octogenarian male patient with acute pyelonephritis. The isolate was found to produce CTX-M-2-type extended-spectrum β-lactamase. Interestingly, the isolate failed to grow on modified Drigalski (BTB) and MacConkey agar media, even under CO2-enriched atmosphere. Our analysis revealed that the pH-indicator dyes, bromothymol blue, and/or crystal violet that were incorporated into the agar media inhibited the growth of the isolate. Although routine identification methods using Vitek® 2 Compact systems were unsuccessful, the isolate was identified as Proteus mirabilis by 16S rRNA sequencing and MALDI-TOF MS analysis. The carbonic anhydrase (CA) region spanning approximately 2,000 bp upstream to 2,000 bp downstream, which is responsible for the CO2 requirement, was not amplified, which could be attributed to the large-scale deletion or mutation of the DNA sequences containing the CA gene region. In fact, revertants with the ability to grow without CO2 were not detected. However, a revertant that was capable of growing in both BTB and MacConkey agar was detected at frequencies less than 10-9. Therefore, the genes responsible for the highly sensitive reactions of the isolate to pH indicator dyes is not likely to be linked to the CA genes.
Mucormycosis is an opportunistic infection occurring in immunocompromised hosts with hematological malignancies. Mortality due to mucormycosis in patients with hematological malignancies is high. However, the clinical symptoms of mucormycosis are poorly characterized, and diagnosis is difficult due to the lack of specific culture or serological markers or antigens. We present two cases in which nested polymerase chain reaction with specific primers was used in the serum of patients with hematological malignancies.
Anaplasma phagocytophilum is an obligate intracellular bacterium that causes human granulocytic anaplasmosis (HGA), an emerging tick-borne infectious disease. This bacterium expresses various 44-kDa major outer membrane proteins encoded by the p44/msp2 multigene family to avoid the host immune system. We previously detected A. phagocytophilum p44/msp2 from the tick Haemaphysalis longicornis in Mie Prefecture, Japan in 2008. In this study, we further investigated a total of 483 H. longicornis ticks (220 adults and 263 nymphs) collected from the Mie Prefecture by PCR targeting p44/msp2 to characterize the p44/msp2 multigene family of A. phagocytophilum. Six of the 483 ticks tested were PCR-positive for A. phagocytophilum p44/msp2, and these positive individuals were at the nymph stage of the tick life cycle. Cloning, sequencing, and phylogenetic analyses of the amplicons revealed that the 11 p44/msp2 clones obtained from the positive ticks shared a 54.9%–99.3% amino acid sequence similarity with the 27 previously identified clones from HGA patients in Japan. In particular, 6 p44/msp2 clones displayed the highest similarities (97.2%–99.3%) with 3 previously identified clones (FJ417343, FJ417345, FJ417357). Thus, the data from this study provide important public health information regarding A. phagocytophilum infection transmitted by H. longicornis ticks, especially at the nymph stage.