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Kitajima M.,University of Arizona | Hata A.,University of Tokyo | Yamashita T.,Aichi Prefectural Institute of Public Health | Haramoto E.,Yamanashi University | And 2 more authors.
Applied and Environmental Microbiology | Year: 2013

Aichi viruses (AiVs) have been proposed as a causative agent of human gastroenteritis potentially transmitted by fecal-oral routes through contaminated food or water. In the present study, we developed a TaqMan minor groove binder (MGB)-based reverse transcription-quantitative PCR (RT-qPCR) system that is able to quantify AiVs and differentiate between genotypes A and B. This system consists of two assays, an AiV universal assay utilizing a universal primer pair and a universal probe and a duplex genotype-specific assay utilizing the same primer pair and two genotype-specific probes. The primers and probes were designed based on multiple alignments of the 21 available AiV genome sequences containing the capsid gene. Using a 10-fold dilution of plasmidDNAcontaining the target sequences, it was confirmed that both assays allow detection and quantification of AiVs with a quantitative range of 1.0×101 to 1.0×107 copies/reaction, and the genotype-specific assay reacts specifically to each genotype. To validate the newly developed assays, 30 clinical stool specimens were subsequently examined with the assays, and the AiVRNA loads were determined to be 1.4×104 to 6.6×109 copies/g stool.Wealso examined 12 influent and 12 effluent wastewater samples collected monthly for a 1-year period to validate the applicability of the assays for detection of AiVs in environmental samples. The AiV RNAconcentrations in influent and effluent wastewater were determined to be up to 2.2×107 and 1.8×104 copies/liter, respectively. Our RT-qPCR system is useful for routine diagnosis of AiVs in clinical stool specimens and environmental samples. © 2013, American Society for Microbiology.


Suzuki M.,Aichi Prefectural Institute of Public Health | Suzuki M.,Nagoya University | Hosoba E.,National Hospital Organization Nagoya Medical Center | Matsui M.,Japan National Institute of Infectious Diseases | Arakawa Y.,Nagoya University
Journal of Clinical Microbiology | Year: 2014

Antimicrobial resistance issues have become a global health concern. The rapid identification of multidrug-resistant microbes, which depends on microbial genomic information, is essential for overcoming growing antimicrobial resistance challenges. However, genotyping methods, such as multilocus sequence typing (MLST), for identifying international epidemic clones of Acinetobacter baumannii are not easily performed as routine tests in ordinary clinical laboratories. In this study, we aimed to develop a novel genotyping method that can be performed in ordinary microbiology laboratories. Several open reading frames (ORFs) specific to certain bacterial genetic lineages or species, together with their unique distribution patterns on the chromosomes showing a good correlation with the results of MLST, were selected in A. baumannii and other Acinetobacter spp. by comparing their genomic data. The distribution patterns of the ORFs were visualized by agarose gel electrophoresis after multiplex PCR amplification and digitized. A. baumannii sequence types (STs) corresponding to international clones I and II were successfully discriminated from other STs and Acinetobacter species by detecting the distribution patterns of their ORFs using the multiplex PCR developed here. Since bacterial STs can be easily expressed as digitized numeric data with plus (+) expressed as 1 and minus (-) expressed as 0, the results of the method can be easily compared with those obtained by different tests or laboratories. This PCR-based ORF typing (POT) method can easily and rapidly identify international epidemic clones of A. baumannii and differentiate this microbe from other Acinetobacter spp. Since this POT method is easy enough to be performed even in ordinary clinical laboratories, it would also contribute to daily infection control measures and surveillance. Copyright © 2014, American Society for Microbiology. All Rights Reserved.


Ogata K.,Oita Prefectural Institute of Health and Environment | Ogata K.,University of Occupational and Environmental Health Japan | Narimatsu H.,Oita Prefectural Institute of Health and Environment | Suzuki M.,Aichi Prefectural Institute of Public Health | And 3 more authors.
Applied and Environmental Microbiology | Year: 2012

The incidence of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infection has been increasing; however, the sources of infection remain unclear. Therefore, we investigated the involvement of meat as a possible mediator of CA-MRSA infection.Weexamined the distribution ofMRSAstrains in commercially distributed raw meat samples (n=197) and diarrheal stool samples of outpatients (n=1,287) that were collected in Oita Prefecture, Japan, between 2003 and 2009 for routine legal inspections. FourteenMRSAstrains were isolated from three meat and 11 stool samples. Among these, seven isolates from three meat and four stool samples exhibited the same epidemiological marker profiles [coagulase type III, staphylococcal enterotoxin C, staphylococcal chromosomal cassette mec (SCCmec) type IV, ST8, spa type 606 (t1767), and toxic shock syndrome toxin-1 (TSST-1) producing type]. Furthermore, of the seven strains, three isolates from two meat samples and one stool sample collected in 2007 exhibited completely identical characteristics with respect to phage open reading frame (ORF) typing, pulsed-field gel electrophoresis, and drug susceptibility profiles. The results suggest that commercially distributed meat could play a role in the prevalence of CA-MRSA in the community. © 2012, American Society for Microbiology.


Suzuki M.,Aichi Prefectural Institute of Public Health | Yamada K.,Aichi Prefectural Institute of Public Health | Nagao M.,Kyoto University | Aoki E.,National Hospital Organization Nagoya Medical Center | And 7 more authors.
Emerging Infectious Diseases | Year: 2011

We tested 259 methicillin-resistant Staphylococcus aureus isolates and 2 USA300 ATCC type strains for susceptibility to bacitracin and neomycin contained in overthe- counter antibacterial ointments. Resistance to both bacitracin and neomycin was found only in USA300. The use of over-the counter antimicrobial drugs may select for the USA300 clone.


Nagao M.,Kyoto University | Iinuma Y.,Kyoto University | Suzuki M.,Aichi Prefectural Institute of Public Health | Matsushima A.,Kyoto University | And 3 more authors.
American Journal of Infection Control | Year: 2010

This report describes the first outbreak of methicillin-resistant Staphylococcus aureus USA300 in a general hospital ward in Japan, involving 6 health care workers and 4 patients. This report emphasizes the need for health care personnel to be alert that methicillin-resistant Staphylococcus aureus harboring Panton-Valentine leukocidin gene poses a threat for both nosocomial and occupational infection. Copyright © 2010 by the Association for Professionals in Infection Control and Epidemiology, Inc., and the Society for Healthcare. Epidemiology of America.


Sueta A.,Aichi Cancer Center Research Institute | Sueta A.,Kumamoto University | Ito H.,Aichi Cancer Center Research Institute | Kawase T.,Aichi Cancer Center Research Institute | And 10 more authors.
Breast Cancer Research and Treatment | Year: 2012

Genome-wide association studies (GWASs) have identified genetic variants associated with breast cancer. Most GWASs to date have been conducted in women of European descent, however, and the contribution of these variants as predictors in Japanese women is unknown. Here, we analyzed 23 genetic variants identified in previous GWASs and conducted a case-control study with 697 case subjects and 1,394 age- and menopausal status-matched controls. We fit conditional regression models with genetic variants and conventional risk factors. In addition, we created a polygenetic risk score, using those variants with a statistically significant association with breast cancer risk, and also evaluated the contribution of these genetic predictors using the c statistic. Eleven single-nucleotide polymorphisms (SNPs) revealed significant associations with breast cancer risk. A dose-dependent association was observed between the risk of breast cancer and the genetic risk score, which was an aggregate measure of alleles in seven selected variants, namely FGFR2-rs2981579, TOX3/TNRC9-rs3803662, C6orf97-rs2046210, 8q24-rs13281615, SLC4A7-rs4973768, LSP1-rs38137198, and CASP8-rs10931936. Compared to women with scores of 3 or less, odds ratios (ORs) for women with scores of 4-5, 6-7, 8-9, and 10 or more were 1.33 (95% confidence interval, 1.00-1.80), 1.71 (1.26-2.30), 3.01 (1.97-4.58), and 8.69 (2.75-27.5), respectively (P trend = 1.9 × 10 -9). The c statistic for a model including the genetic risk score in addition to the conventional risk factors was 0.6933, versus 0.6652 with the conventional risk factors only (P = 1.3 × 10 -4). Population-attributable fraction of the risk score was 33.0%. In conclusion, we identified a genetic risk predictor of breast cancer in a Japanese population. Risk models which include a genetic risk score are possibly useful in distinguishing women at high risk of breast cancer from those at low risk, particularly in the context of targeted prevention. © 2011 Springer Science+Business Media, LLC.


Goto T.,Aichi Prefectural Institute of Public Health
Yakugaku Zasshi | Year: 2010

One of the major roles of public health agencies is to ensure safe products for consumers through analysis of residual agricultural chemicals and veterinary drugs in foods. The use of agricultural chemicals and veterinary drugs in agriculture is necessary to improve the quality of the food produced. They play a beneficial role in providing a plentiful, low-cost supply of high-quality food. On the other hand, as a consequence of this use, the presence of residues in food that is a critical element of overall public health is unavoidable, and residues in food are of great importance in the evaluation of food quality. N-methyl carbamate pesticides have anticholinesterase activity and are used worldwide. The postcolumn HPLC method is widely used for the detection of N-methyl carbamate pesticides in food. However, this traditional method involves a time-consuming process that requires skillful techniques. Therefore we developed a new analysis method for N-methyl carbamate pesticides as described in detail in this paper. The new method, including sample preparation and determination, is simple and rapid and allows simultaneous determination of pesticides in food within a much shorter analysis time as compared with the traditional method. This should provide high-quality analysis and ensure safe products for consumers. © 2010 The Pharmaceutical Society of Japan.


Kobayashi S.,Aichi Prefectural Institute of Public Health | Fujiwara N.,Aichi Prefectural Institute of Public Health | Yasui Y.,Aichi Prefectural Institute of Public Health | Yamashita T.,Aichi Prefectural Institute of Public Health | And 2 more authors.
Archives of Virology | Year: 2012

Sapovirus (SaV) is a common cause of acute viral gastroenteritis worldwide, and SaV outbreaks have become more frequent in recent years. In January 2010, an outbreak of acute gastroenteritis due to SaV occurred in Aichi, Gifu and Mie Prefectures, Japan. The illness was strongly associated with eating a delivered box lunch prepared by one catering company. In total, 655 (17.1 %) of 3827 individuals developed gastroenteritic symptoms. SaV was detected in seven of the nine people who became ill and in seven of the 52 food handlers at the catering company, but all the tested samples were negative for norovirus and enteropathogenic bacteria. Sequence analysis of RT-PCR products indicated that the nucleotide sequences of SaV strains from the people who became ill and the food handlers were identical. The detected SaV strains were genogrouped as SaV genotype I.2. This was the largest foodborne outbreak of sapovirus in Japan. © 2012 Springer-Verlag.


Ueno E.,Aichi Prefectural Institute of Public Health
Journal of Pesticide Science | Year: 2015

In Japan, the Positive List System for the regulation of agricultural chemical residues in foods has been in force since May 29, 2006. Moreover, food poisoning caused by methamidophos-laced frozen gyoza dumplings came to light on Jan. 30, 2008. The news has set off alarms about the safety of not only perishables but also unexpected pesticides in processed foods. Therefore, a reliable systematic method was developed for determining pesticide residues in various foods. Firstly, ca. 200 target pesticides were selected by statistically analyzing the monitoring data in Aichi Prefecture. Secondly, a systematic method using plural separation and detection systems combining GC-MS/MS and LC-MS/MS as first priority was constructed. As the sample preparation method corresponding to the systematic method, a simple and easy acetonitrile extraction method, an auto-cleanup system combining GPC and mini-column SPE were developed. Thirdly, a new official multi-residue method that is able to determine a wide range of pesticides in various foods including fatty processed foods was developed. And finally, a comprehensive chromatographic detection system by dual-column GC-MS(/MS) and an interactive database without the use of pesticide standards was commercialized. © Pesticide Science Society of Japan.


Ito M.,Aichi Prefectural Institute of Public Health | Yamashita T.,Aichi Prefectural Institute of Public Health | Tsuzuki H.,Aichi Prefectural Institute of Public Health | Kabashima Y.,Aichi Prefectural Institute of Public Health | And 7 more authors.
Journal of Clinical Microbiology | Year: 2010

Between April 1999 and March 2008, a total of 4,976 stool specimens collected from patients with suspected viral infection through infectious agent surveillance in Aichi, Japan, were tested for the presence of human parechoviruses (HPeVs). We detected HPeVs in 110 samples by either cell culture, reverse transcriptase PCR (RT-PCR), or both. Serotyping either by neutralization test or by nucleotide sequence determination and phylogenetic analysis of the VP1 region and 5′ untranslated region (5′UTR) regions revealed that 63 were HPeV type 1 (HPeV-1), followed by 44 HPeV-3 strains, 2 HPeV-4 strains, and 1 HPeV-6 strain. The high nucleotide and amino acid sequence identities of the Japanese HPeV-3 isolates in 2006 to the strains previously reported from Canada and Netherlands confirmed the worldwide prevalence of HPeV-3 infection. Ninety-seven percent of the HPeV-positive patients were younger than 3 years, and 86.2% younger than 12 months. The clinical diagnoses of HPeV-positive patients were gastroenteritis, respiratory illness, febrile illness, exanthema, "hand, foot, and mouth disease," aseptic meningitis, and herpangina. Among 49 HPeV-positive patients with gastroenteritis, 35 were positive with HPeV-1 and 12 with HPeV-3, and out of 25 with respiratory illness, 11 were positive with HPeV-1 and 14 with HPeV-3. HPeV-3 seemed to be an important etiological agent of respiratory infection of children. While HPeV-1 was detected predominantly during fall and winter, the majority of the HPeV-3 cases were detected during summer and fall. A different pattern of clinical manifestations as well as seasonality suggested that there are different mechanisms of pathogenesis between HPeV-1 and HPeV-3 infections. Copyright © 2010, American Society for Microbiology. All Rights Reserved.

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