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Chang B.,Japan National Institute of Infectious Diseases | Taguri T.,Nagasaki Prefectural Institute for Environmental Research and Public Health | Sugiyama K.,Shizuoka Institute of Environment and Hygiene | Amemura-Maekawa J.,Japan National Institute of Infectious Diseases | And 2 more authors.
Japanese Journal of Infectious Diseases | Year: 2010

Ethidium monoazide (EMA) and propidium monoazide (PMA) have been utilized for selective PCR amplification of DNA from viable bacterial cells. In this study, we compared the abilities of EMA and PMA, together with real-time PCR, to specifically distinguish dead Legionella cells from viable cells. Several experiments showed that PMA or EMA treatment could specifically prevent the PCR amplification of DNA from dead Legionella cells in water samples. However, a 4-fold higher concentration of PMA than EMA was required to achieve this effect. EMA may therefore be more useful for practical environmental investigations of Legionella. Source


Taira Y.,Nagasaki University | Taira Y.,Nagasaki Prefectural Institute for Environmental Research and Public Health | Hayashida N.,Nagasaki University | Tsuchiya R.,Nagasaki University | And 8 more authors.
PLoS ONE | Year: 2013

For the current on-site evaluation of the environmental contamination and contributory external exposure after the accident at the Chernobyl Nuclear Power Plant (CNPP) and the nuclear tests at the Semipalatinsk Nuclear Testing Site (SNTS), the concentrations of artificial radionuclides in soil samples from each area were analyzed by gamma spectrometry. Four artificial radionuclides (241Am,134Cs, 137Cs, and 60Co) were detected in surface soil around CNPP, whereas seven artificial radionuclides (241Am, 57Co,137Cs, 95Zr, 95Nb, 58Co, and 60Co) were detected in surface soil around SNTS. Effective doses around CNPP were over the public dose limit of 1 mSv/y (International Commission on Radiological Protection, 1991). These levels in a contaminated area 12 km from Unit 4 were high, whereas levels in a decontaminated area 12 km from Unit 4 and another contaminated area 15 km from Unit 4 were comparatively low. On the other hand, the effective doses around SNTS were below the public dose limit. These findings suggest that the environmental contamination and effective doses on the ground definitely decrease with decontamination such as removing surface soil, although the effective doses of the sampling points around CNPP in the present study were all over the public dose limit. Thus, the remediation of soil as a countermeasure could be an extremely effective method not only for areas around CNPP and SNTS but also for areas around the Fukushima Dai-ichi Nuclear Power Plant (FNPP), and external exposure levels will be certainly reduced. Long-term follow-up of environmental monitoring around CNPP, SNTS, and FNPP, as well as evaluation of the health effects in the population residing around these areas, could contribute to radiation safety and reduce unnecessary exposure to the public. © 2013 Taira et al. Source


Yoshikawa T.,Japan National Institute of Infectious Diseases | Fukushi S.,Japan National Institute of Infectious Diseases | Tani H.,Japan National Institute of Infectious Diseases | Fukuma A.,Japan National Institute of Infectious Diseases | And 25 more authors.
Journal of Clinical Microbiology | Year: 2014

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease with a high case fatality risk and is caused by the SFTS virus (SFTSV). A retrospective study conducted after the first identification of an SFTS patient in Japan revealed that SFTS is endemic to the region, and the virus exists indigenously in Japan. Since the nucleotide sequence of Japanese SFTSV strains contains considerable differences compared with that of Chinese strains, there is an urgent need to establish a sensitive and specific method capable of detecting the Chinese and Japanese strains of SFTSV. A conventional one-step reverse transcription-PCR (RT-PCR) (cvPCR) method and a quantitative one-step RT-PCR (qPCR) method were developed to detect the SFTSV genome. Both cvPCR and qPCR detected a Chinese SFTSV strain. Forty-one of 108 Japanese patients suspected of having SFTS showed a positive reaction by cvPCR. The results from the samples of 108 Japanese patients determined by the qPCR method were in almost complete agreement with those determined by cvPCR. The analyses of the viral copy number level in the patient blood samples at the acute phase determined by qPCR in association with the patient outcome confirmed that the SFTSV RNA load in the blood of the nonsurviving patients was significantly higher than that of the surviving patients. Therefore, the cvPCR and qPCR methods developed in this study can provide a powerful means for diagnosing SFTS. In addition, the detection of the SFTSV genome level by qPCR in the blood of the patients at the acute phase may serve as an indicator to predict the outcome of SFTS. Copyright © 2014, American Society for Microbiology. All Rights Reserved. Source


Yoshikawa T.,Japan National Institute of Infectious Diseases | Shimojima M.,Japan National Institute of Infectious Diseases | Fukushi S.,Japan National Institute of Infectious Diseases | Tani H.,Japan National Institute of Infectious Diseases | And 25 more authors.
Journal of Infectious Diseases | Year: 2015

Background. Severe fever with thrombocytopenia syndrome (SFTS) is a tick-borne acute infectious disease caused by the SFTS virus (SFTSV). SFTS has been reported in China, South Korea, and Japan as a novel Bunyavirus. Although several molecular epidemiology and phylogenetic studies have been performed, the information obtained was limited, because the analyses included no or only a small number of SFTSV strains from Japan. Methods. The nucleotide sequences of 75 SFTSV samples in Japan were newly determined directly from the patients' serum samples. In addition, the sequences of 7 strains isolated in vitro were determined and compared with those in the patients' serum samples. More than 90 strains that were identified in China, 1 strain in South Korea, and 50 strains in Japan were phylogenetically analyzed. Results. The viruses were clustered into 2 clades, which were consistent with the geographic distribution. Three strains identified in Japan were clustered in the Chinese clade, and 4 strains identified in China and 26 in South Korea were clustered in the Japanese clade. Conclusions. Two clades of SFTSV may have evolved separately over time. On rare occasions, the viruses were transmitted overseas to the region in which viruses of the other clade were prevalent. © 2015 The Author 2015. Source


Tsujimura K.,Nagasaki Prefectural Institute for Environmental Research and Public Health | Yamanouchi K.,Nagasaki Prefectural Institute for Environmental Research and Public Health
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2015

A simple and rapid detection method for tetrodotoxin (TTX), a powerful sodium channel blocker, in small volumes of the serum of patients with pufferfish poisoning, was developed using an ultrafiltration spin column. The separation and identification of TTX was performed by liquid chromatography (LC) with a multi-mode ODS column and tandem mass spectrometry. TTX and an internal standard (voglibose) were monitored and quantitated using ion transitions: the respective precursor-to-product ion combinations, m/z 320/162 for TTX and 268/92 in MRM mode. The recoveries of TTX and voglibose were 91.0–110.8% and 104.7–107.4%, respectively, and with high accuracy (intra-run, 4.35–5.29%; inter-run, 2.95–5.79%) and linearity (0.5–200 ng/ml serum: r = 0.9994). The lower limit of quantification was 0.5 ng/ml serum. In patients, maximum serum TTX concentrations were 30.2 ng/ml serum for patient 1 on day 0 and 56.1 ng/ml serum for patient 2 on day 1. These results are important for the treatment of patients and for the identification of poisoning as well as for the determination of the cause of the food poisoning. © 2015, © 2015 Taylor & Francis. Source

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