Hiroshi Ushijima

Publications of Hiroshi Ushijima

• Emergence of a new norovirus GII.6 variant in Japan, 2008-2009.

  Authors: Wisoot Chan-It, Aksara Thongprachum, Pattara Khamrin, Masaaki Kobayashi, Shoko Okitsu, Masashi Mizuguchi, Hiroshi Ushijima

  Journal of medical virology. 07/2012; 84(7):1089-96.

  Norovirus (NoV) is recognized as one of the most common causative agents of diarrhea disease in young children. A total of 187 fecal specimens collected from non-hospitalized children with acuteNorovirus (NoV) is recognized as one of the most common causative agents of diarrhea disease in young children. A total of 187 fecal specimens collected from non-hospitalized children with acute gastroenteritis in Shizuoka, Japan during July 2008 to June 2009 were investigated for the presence of diarrhea viruses by a multiplex RT-PCR. Diarrhea viruses were overall detected in 158 of 187 (84.5%). Of the viruses detected, NoV was the most prevalent (55.6%). Most of the NoV sequences belonged to GII.4 (53.8%). NoV GII.6 emerged as the second most common strain (40.4%). The full-length capsid sequences of five representative Shizuoka GII.6 strains were compared with all 12 GII.6 strains available in GenBank database between 1990 and 2009. At least three distinct GII.6 subclusters (a-c) appeared in different parts of the world. Shizuoka GII.6 strains formed their own subcluster c, distinct from other complete GII.6 reference sequences. The Shizuoka strains had significant amino acid divergence, particularly in the P2 domain up to 10.9-17.5% and contained eight unique mutations in the P domains, compared with subcluster a and b viruses. The homology model showed that the eight mutations were predicted to be located at the surface-exposed P1 and P2 domains. The data suggest the emergence of a new NoV GII.6 variant in Shizuoka, with a high level of genetic variation. J. Med. Virol. 84: 1089-1096, 2012. © 2012 Wiley Periodicals, Inc.

• Detection of human bocavirus 1 and 2 from children with acute gastroenteritis in Japan.

  Authors: Pattara Khamrin, Aksara Thongprachum, Hiroyuki Shimizu, Shoko Okitsu, Masashi Mizuguchi, Satoshi Hayakawa, Niwat Maneekarn, Hiroshi Ushijima

  Journal of medical virology. 06/2012; 84(6):901-5.

  This study reports the detection and the genetic characterization of HBoV, a new member in the Parvoviridae family. Among 177 fecal specimens collected from children with diarrhea younger than 5This study reports the detection and the genetic characterization of HBoV, a new member in the Parvoviridae family. Among 177 fecal specimens collected from children with diarrhea younger than 5 years in Japan, 11 (6.2%) were positive for HBoV. Co-infection with other enteric viruses (norovirus and adenovirus) was found in nine (81.8%) pediatric patients, while monoinfection by HBoV alone was detected in two (18.2%) cases. Nucleotide sequence analysis of 11 Japanese HBoV strains revealed that seven HBoV sequences were most closely related to other HBoV 1 reference strains, while the other four strains were similar to HBoV 2A. These results indicated that HBoV was one of the viral agents found in stool specimens collected from children with acute gastroenteritis and HBoV 1 and 2A circulated in Japan between 2009 and 2010 epidemic season. J. Med. Virol. 84:901-905, 2012. © 2012 Wiley Periodicals, Inc.

• Genotypic linkages of gene segments of rotaviruses circulating in pediatric patients with acute gastroenteritis in Thailand.

  Authors: Natthawan Chaimongkol, Pattara Khamrin, Rungnapa Malasao, Aksara Thongprachum, Hiroshi Ushijima, Niwat Maneekarn

  Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases. 05/2012;

  Rotavirus is a major cause of morbidity and mortality of infants and young children with diarrhea throughout the world. In Thailand, extensive studies of rotavirus infections have been reportedRotavirus is a major cause of morbidity and mortality of infants and young children with diarrhea throughout the world. In Thailand, extensive studies of rotavirus infections have been reported continually and rotavirus diarrhea remains a common illness. To monitor the epidemiological situation of rotavirus in Chiang Mai, Thailand, surveillance of rotavirus circulating in pediatric patients was conducted. A total of 160 fecal specimens collected from children hospitalized with diarrhea were tested for rotaviruses groups A, B, and C by RT-PCR and their genotypes were identified by multiplex PCR and nucleotide sequencing. Group A rotavirus was detected at 29.4% but none of group B and C was found in this study. Molecular characterizations of G- and P-genotypes revealed three different G-P combinations, G1P[8] was the most predominant genotype with the prevalence of 72.3% followed by G2P[4] at 19.2%, and G3P[8] at 8.5%. Phylogenetic analyses of VP7 and VP4 genes of the representative strains detected in the present study, G1, G2, G3, and P[4] and P[8], respectively, revealed that G1 belonged to G1-Ic and G1-II, G2 belonged to G2-II, and G3 belonged to G3-III-S4 lineages while P[4] and P[8] were identified as P[4]-V and P[8]-III lineages. Analyses of VP6, NSP4, and NSP5 genes demonstrated that these representative strains belonged to genotypes I1 and I2, E1 and E2, and H1 and H2, respectively. Analyzing the association of G- and P-genotypes with I, E, H genotypes revealed unique patterns of genotypic linkage. The G1P[8] and G3P[8] were intimately linked with I1, E1, H1 genotypes and displayed the genetic features of G1-P[8]-I1-E1-H1 and G3-P[8]-I1-E1-H1, respectively, while G2P[4] was closely linked to I2, E2, H2 genotypes and showed the genetic pattern of G2-P[4]-I2-E2-H2. This study provides epidemiological information and insight into the genetic background of rotaviruses circulating in pediatric patients in Chiang Mai, Thailand.

• Novel human adenovirus strain, bangladesh.

  Authors: Yuki Matsushima, Hideaki Shimizu, Atsuko Kano, Etsuko Nakajima, Yoko Ishimaru, Shuvra Kanti Dey, Yuki Watanabe, Fuyuka Adachi, Keiichiro Suzuki, Kohnosuke Mitani, Tsuguto Fujimoto, Tung Gia Phan, Hiroshi Ushijima

  Emerging infectious diseases. 05/2012; 18(5):846-8.

  We report a novel human adenovirus D (HAdV-65) isolated from feces of 4 children in Bangladesh who had acute gastroenteritis. Corresponding genes of HAdV-65 were related to a hexon gene of HAdV-10,We report a novel human adenovirus D (HAdV-65) isolated from feces of 4 children in Bangladesh who had acute gastroenteritis. Corresponding genes of HAdV-65 were related to a hexon gene of HAdV-10, penton base genes of HAdV-37 and HAdV-58, and a fiber gene of HAdV-9. This novel virus may be a serious threat to public health.

• Novel Human Adenovirus Strain, Bangladesh

  Authors: Yuki Matsushima, Hideaki Shimizu, Atsuko Kano, Etsuko Nakajima, Yoko Ishimaru, Shuvra Kanti Dey, Yuki Watanabe, Fuyuka Adachi, Keiichiro Suzuki, Kohnosuke Mitani, Tsuguto Fujimoto, Tung Gia Phan, Hiroshi Ushijima

  Emerging Infectious Diseases. 05/2012; 18(5):846-848.

  We report a novel human adenovirus D (HAdV-65) isolated from feces of 4 children in Bangladesh who had acute gastroenteritis. Corresponding genes of HAdV-65 were related to a hexon gene of HAdV-10,We report a novel human adenovirus D (HAdV-65) isolated from feces of 4 children in Bangladesh who had acute gastroenteritis. Corresponding genes of HAdV-65 were related to a hexon gene of HAdV-10, penton base genes of HAdV-37 and HAdV-58, and a fi ber gene of HAdV- 9. This novel virus may be a serious threat to public health.

• Phylogenetic analysis of rubella viruses in Vietnam during 2009-2010.

  Authors: Dinh Nguyen Tran, Ngan Thi Kim Pham, Thi Thuy Trinh Tran, Pattara Khamrin, Aksara Thongprachum, Katsuhiro Komase, Satoshi Hayakawa, Masashi Mizuguchi, Hiroshi Ushijima

  Journal of medical virology. 04/2012; 84(4):705-10.

  Rubella virus (RV) usually causes a mild disease. However, infection during the first trimester of pregnancy often leads to severe birth defects known as congenital rubella syndrome (CRS). AlthoughRubella virus (RV) usually causes a mild disease. However, infection during the first trimester of pregnancy often leads to severe birth defects known as congenital rubella syndrome (CRS). Although wild-type RVs exist and circulate worldwide, their genotypes remain unknown in many countries. The aim of this study was to identify the molecular characteristics of RVs found in Vietnam during the years 2009-2010 and to provide the first data concerning RV genotypes in this country. Throat swab samples were collected between 2009 and 2010 from four CRS cases and nine rubella infection cases visiting one Children's Hospital and one outpatient clinic in Ho Chi Minh City. The 739-nucleotide coding region of the RV E1 gene recommended by the World Health Organization was amplified by reverse transcriptase PCR, and the resulting DNA fragments were then sequenced. Sequences were assigned to genotypes by phylogenetic analysis with RV reference strains. RV RNA was detected in 11 clinical specimens. Phylogenetic analysis of the sequences showed that all 11 strains belonged to 2B genotype. Several variations in amino acids were found, among which five changes were involved in the B and T cell epitopes. These data indicate that viruses of genotype 2B were circulating in Vietnam. The increasing information about RV genotype in Vietnam should aid in the control of rubella infection and CRS in this country.

• Drug Resistance Mutations in the HIV Type 1 Protease and Reverse Transcriptase Genes in Antiretroviral-Naive Vietnamese Children.

  Authors: Quang Duy Trinh, Ngan Thi Kim Pham, Nhut Tin Le Nguyen, Bao Quoc Lam, Kim Thoa Le Phan, Khanh Huu Truong, Yasuyuki Izumi, Shihoko Komine-Aizawa, Masashi Mizuguchi, Hiroshi Ushijima, Satoshi Hayakawa

  AIDS research and human retroviruses. 03/2012;

  Abstract Anti-HIV drugs have recently become available for the treatment of children infected with HIV in Vietnam; however, the genetic background of HIV-1 drug resistance in antiretroviral-naiveAbstract Anti-HIV drugs have recently become available for the treatment of children infected with HIV in Vietnam; however, the genetic background of HIV-1 drug resistance in antiretroviral-naive children has yet to be studied. Of the 104 HIV-1 CRF01-AE subtype strains that were previously isolated from antiretroviral-naive children from the provinces of southern Vietnam and hospitalized in Children Hospital 1 in Ho Chi Minh City from 2004 to 2005, 79 strains were used for amplification and sequence analyses of the protease and reverse transcriptase (RT) genes. Minor mutations were found in the protease gene, including L10I, I13V, G16E, M36I, D60E, I62V, I64V, L63P, H69K, V82I, and I93L. Of these mutations, M36I and H69K were detected in all of the strains that were studied. However, all of the amino acid changes in the protease gene were considered to be polymorphisms. In the RT gene, three major mutations were detected in six strains: the V75M mutation in one strain, the Y181C mutation in two strains, and the M184I mutation in three strains. The prevalence of primary or transmitted HIV drug resistance to all of the drugs and drug classes that were evaluated in this study was 7.6%. These findings provide a useful background for antiretroviral therapy in Vietnam and contribute reference data for the surveillance of HIV drug resistance around the world. This study suggests that the prevalence of HIVDR in Vietnam may have recently increased. The monitoring of HIV drug resistance in Vietnam is necessary.

• Analysis of base and codon usage by rubella virus.

  Authors: Yumei Zhou, Xianfeng Chen, Hiroshi Ushijima, Teryl K Frey

  Archives of virology. 02/2012;

  Rubella virus (RUBV), a small, plus-strand RNA virus that is an important human pathogen, has the unique feature that the GC content of its genome (70%) is the highest (by 20%) among RNA viruses. ToRubella virus (RUBV), a small, plus-strand RNA virus that is an important human pathogen, has the unique feature that the GC content of its genome (70%) is the highest (by 20%) among RNA viruses. To determine the effect of this GC content on genomic evolution, base and codon usage were analyzed across viruses from eight diverse genotypes of RUBV. Despite differences in frequency of codon use, the favored codons in the RUBV genome matched those in the human genome for 18 of the 20 amino acids, indicating adaptation to the host. Although usage patterns were conserved in corresponding genes in the diverse genotypes, within-genome comparison revealed that both base and codon usages varied regionally, particularly in the hypervariable region (HVR) of the P150 replicase gene. While directional mutation pressure was predominant in determining base and codon usage within most of the genome (with the strongest tendency being towards C's at third codon positions), natural selection was predominant in the HVR region. The GC content of this region was the highest in the genome (>80%), and it was not clear if selection at the nucleotide level accompanied selection at the amino acid level. Dinucleotide frequency analysis of the RUBV genome revealed that TpA usage was lower than expected, similar to mammalian genes; however, CpG usage was not suppressed, and TpG usage was not enhanced, as is the case in mammalian genes.

• Circulating of human bocavirus 1, 2, 3, and 4 in pediatric patients with acute gastroenteritis in Thailand.

  Authors: Pattara Khamrin, Rungnapa Malasao, Natthawan Chaimongkol, Nuthapong Ukarapol, Tipachan Kongsricharoern, Shoko Okitsu, Satoshi Hayakawa, Hiroshi Ushijima, Niwat Maneekarn

  Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases. 02/2012; 12(3):565-9.

  Human bocavirus (HBoV) is a newly identified human parvovirus that associated with respiratory and gastrointestinal diseases. Epidemiological surveillance of HBoV was conducted on fecal specimensHuman bocavirus (HBoV) is a newly identified human parvovirus that associated with respiratory and gastrointestinal diseases. Epidemiological surveillance of HBoV was conducted on fecal specimens collected from hospitalized children with diarrhea in Chiang Mai, Thailand in 2011. Among a total of 222 fecal specimens tested, 17 (7.7%) were positive for HBoV by PCR. Of the 17 HBoV positive samples, double- or triple-infections together with other enteric viruses were found in 10 (58.8%) pediatric patients, while monoinfection with HBoV alone was detected in seven (41.2%) cases. Mixed infection among HBoV with norovirus GII was frequently observed in this population. The partial VP1 nucleotide sequences of all 17 HBoV strains demonstrated that all four species of HBoV were found in the specimens tested. Eleven strains were HBoV1. Other three strains showed the sequence identity with HBoV2, which were most closely related to the HBoV2A. In addition, other two HBoV strains showed the highest level of nucleotide sequence identity with the HBoV3. It was surprisingly to observe that one Thai HBoV strain showed a unique characteristic similar to the HBoV4, a rare species of HBoV found in acute gastroenteritis patients. In summary, this study presents the genetic background information of HBoV circulated in acute gastroenteritis children in Chiang Mai, Thailand and it was clearly demonstrated that HBoVs circulated in this area were genetically diverse as all four species of HBoVs (HBoV1-4) were detected in the fecal specimens collected from pediatric patients admitted to the hospitals in this area.

• A wide variety of diarrhea viruses circulating in pediatric patients in Thailand.

  Authors: Natthawan Chaimongkol, Pattara Khamrin, Boonpa Suantai, Wilaiporn Saikhreang, Aksara Thongprachum, Rungnapa Malasao, Nuthapong Ukarapol, Tipachan Kongsricharoern, Hiroshi Ushijima, Niwat Maneekarn

  Clinical laboratory. 01/2012; 58(1-2):117-23.

  Viral gastroenteritis has been recognized as one of the most common illnesses that affects infants and young children all over the world. A wide variety of viruses associated with the disease areViral gastroenteritis has been recognized as one of the most common illnesses that affects infants and young children all over the world. A wide variety of viruses associated with the disease are continually being reported. To investigate the epidemiological situation of diarrhea virus infection in Chiang Mai, Thailand, surveillance was conducted during January to December 2007. A total of 160 fecal specimens collected from pediatric patients admitted to the hospital with acute gastroenteritis were tested for the presence of group A, B, and C rotaviruses, norovirus, sapovirus, astrovirus, adenovirus, Aichi virus, enterovirus, bocavirus, and human parechovirus by RT-multiplex PCR. Of 160 fecal specimens tested, 85 (53.1%) were positive for diarrhea viruses. Of these, group A rotavirus was the predominant with a prevalence of 27.5%, followed by norovirus GII (11.9%), sapovirus (3.1%), enterovirus (2.5%), human parechovirus (1.9%), and norovirus GI, astrovirus, adenovirus (each 0.6%). Mixed-infections of 2 or 3 viruses were observed in 7 (4.4%) patients. However, none of groups B and C rotaviruses and Aichi virus were detected in this study. Monthly distribution analysis revealed that all those diarrhea viruses were detected continually throughout the year at a low level of infection except for group A rotavirus and norovirus infections which appeared to peak in a cool season in January-March and December, respectively. This surveillance revealed a wide variety of diarrhea viruses currently circulating in pediatric patients with acute gastroenteritis in Chiang Mai, Thailand.

• Detection and molecular characterization of cosavirus in adults with diarrhea, Thailand.

  Authors: Pattara Khamrin, Natthawan Chaimongkol, Rungnapa Malasao, Boonpa Suantai, Wilaiporn Saikhruang, Tipachan Kongsricharoern, Nuthapong Ukarapol, Shoko Okitsu, Hiroyuki Shimizu, Satoshi Hayakawa, Hiroshi Ushijima, Niwat Maneekarn

  Virus genes. 12/2011; 44(2):244-6.

  Human cosavirus (HCoSV) is a newly discovered virus in Picornaviridae family. At present it is not clear whether HCoSV is associated with diseases, including gastroenteritis in humans, asHuman cosavirus (HCoSV) is a newly discovered virus in Picornaviridae family. At present it is not clear whether HCoSV is associated with diseases, including gastroenteritis in humans, as epidemiological data is limited. Epidemiological surveillance of HCoSV was conducted on 150 fecal specimens collected from children and 150 samples from adults with diarrhea in Thailand by RT-PCR screening. HCoSV was found in a single adult specimen and not in any of the fecal specimens from children. This represents the first report of HCoSV infection in patients with diarrhea in Thailand. Extensive epidemiological surveillance of novel viruses associated with diarrhea in other populations may provide a better understanding of the distribution, genetic diversity, and association of the viral agents associated with acute gastroenteritis in humans.

• Sequence analysis of porcine kobuvirus VP1 region detected in pigs in Japan and Thailand.

  Authors: Shoko Okitsu, Pattara Khamrin, Aksara Thongprachum, Satoshi Hidaka, Sompreeya Kongkaew, Apisek Kongkaew, Niwat Maneekarn, Masashi Mizuguchi, Satoshi Hayakawa, Hiroshi Ushijima

  Virus genes. 11/2011; 44(2):253-7.

  Porcine kobuvirus is a new candidate species of the genus Kobuvirus in the family Picornaviridae, and information is still limited. The identification of porcine kobuvirus has been performed by thePorcine kobuvirus is a new candidate species of the genus Kobuvirus in the family Picornaviridae, and information is still limited. The identification of porcine kobuvirus has been performed by the sequence analyses of the 3D region of the viruses. Therefore, the purpose of this study was to characterize the molecular properties of VP1 nucleotide sequences of the porcine kobuviruses isolated from porcine stool samples in Japan during 2009 and Thailand between 2006 and 2008. In addition, previous identification of a unique porcine kobuvirus; Japanese H023/2009/JP, which is a bovine kobuvirus-like strain based on sequence analysis of the 3D region, was also included in this study. All of the strains were amplified by the VP1-specific primer pair: the amplicons were subjected to direct sequencing and compared with the VP1 nucleotide sequences of reference strains. The VP1 sequences of strains from the GenBank database revealed high nucleotide sequence identity at 84.3-100%. On the other hand, the nucleotide identities among the 15 porcine kobuvirus strains analyzed in this study ranged from 78.8 to 99.8%. The results revealed that diversity of the strains in this study were higher than those of the strains in previous studies. Furthermore, it was found that the VP1 region of the bovine kobuvirus-like strain, H023/2009/JP, clustered with nine porcine kobuvirus strains that were isolated in Thailand and Japan. Since this strain was previously found to be closely related to bovine kobuviruses in the 3D gene region, it may be a natural recombinant.

• Genomic characterization of a novel human adenovirus type 31 recombinant in the hexon gene.

  Authors: Yuki Matsushima, Hideaki Shimizu, Tung Gia Phan, Hiroshi Ushijima

  The Journal of general virology. 08/2011; 92(Pt 12):2770-5.

  A novel human recombinant adenovirus of species A (HAdV-A31 MZ) was isolated from a patient with acute gastroenteritis in Japan. The complete genome of HAdV-A31 strain MZ contains 33 776 bp. AnalysisA novel human recombinant adenovirus of species A (HAdV-A31 MZ) was isolated from a patient with acute gastroenteritis in Japan. The complete genome of HAdV-A31 strain MZ contains 33 776 bp. Analysis of the hexon gene of HAdV-A31 MZ indicated that its hexon sequence is the result of a genetic recombination between those of HAdV-A31 and a close relative to HAdV-A12. The recombination sites were found around the border of hypervariable loops 1 and 2 in the hexon gene, which are the most important determinants for virus neutralization. Loops 1 and 2 of this virus were genetically related to HAdV-A12, whereas all other parts of the genome were highly similar to HAdV-A31. In order to understand the evolution of adenoviruses correctly and to avoid misidentification of HAdV types, we recommend characterizing not only the hexon gene, but also the penton base and fiber genes.

• Saffold cardioviruses in children with diarrhea, Thailand.

  Authors: Pattara Khamrin, Natthawan Chaimongkol, Nattika Nantachit, Shoko Okitsu, Hiroshi Ushijima, Niwat Maneekarn

  Emerging infectious diseases. 06/2011; 17(6):1150-2.

  To the Editor: Cardioviruses currently consist of at least 3 viruses: Theiler murine encephalomyocarditis virus, encephalomyocarditis virus, and Saffold virus (SAFV) (1-4). Saffold cardiovirus in theTo the Editor: Cardioviruses currently consist of at least 3 viruses: Theiler murine encephalomyocarditis virus, encephalomyocarditis virus, and Saffold virus (SAFV) (1-4). Saffold cardiovirus in the family Picornaviridae was isolated and identified from fecal specimens of a child with fever of unknown origin in the United States (3).

• Human bocavirus infection in children with acute gastroenteritis in Japan and Thailand.

  Authors: Ngan Thi Kim Pham, Quang Duy Trinh, Wisoot Chan-It, Pattara Khamrin, Shuichi Nishimura, Kumiko Sugita, Niwat Maneekarn, Shoko Okitsu, Masashi Mizuguchi, Hiroshi Ushijima

  Journal of medical virology. 02/2011; 83(2):286-90.

  A total of 329 fecal specimens, which had been known to be negative for rotavirus, adenovirus, norovirus, sapovirus, and astrovirus, and which were collected from infants and children with acuteA total of 329 fecal specimens, which had been known to be negative for rotavirus, adenovirus, norovirus, sapovirus, and astrovirus, and which were collected from infants and children with acute gastroenteritis in Japan and Thailand during 2005-2008 were screened for human bocavirus (HBoV). HBoV was detected by PCR with a primer pair that amplified the NP1 region of its genome and was genotyped by sequencing of the VP1/VP2 region. Of the 329 samples tested, 6 (1.8%) were positive for HBoV. Of these, five samples were collected from Japan and one sample was from Thailand, and the detection rates of HBoV in each country were 2% and 1.2%, respectively. For the detected HBoV, the capsid VP1/VP2 gene of all HBoV strains was successfully sequenced. Four Japanese HBoV strains studied were clustered into group 1, while the remaining Japanese strain and a unique Thai strain belonged to group 2. No severe acute gastroenteritis associated with HBoV was noted. This study provides better understanding on the epidemiology of HBoV infections in children with acute gastroenteritis in Japan and Thailand.

• Detection of human parechovirus in stool samples collected from children with acute gastroenteritis in Japan during 2007-2008.

  Authors: Ngan Thi Kim Pham, Wisoot Chan-It, Pattara Khamrin, Shuichi Nishimura, Hideaki Kikuta, Kumiko Sugita, Tsuneyoshi Baba, Atsuko Yamamoto, Hideaki Shimizu, Shoko Okitsu, Masashi Mizuguchi, Hiroshi Ushijima

  Journal of medical virology. 02/2011; 83(2):331-6.

  Of 477 stool specimens, which had been screened for rotavirus, adenovirus, norovirus, sapovirus and astrovirus, collected from infants and children with acute gastroenteritis in pediatric clinicsOf 477 stool specimens, which had been screened for rotavirus, adenovirus, norovirus, sapovirus and astrovirus, collected from infants and children with acute gastroenteritis in pediatric clinics encompassing five localities (Sapporo, Tokyo, Maizuru, Osaka, and Saga) in Japan from July 2007 to June 2008, 247 negative samples (51.7%) were subjected to screening for human parechovirus. Human parechovirus (HPeV) was detected by RT-PCR using a primer pair to amplify 5'UTR region of its genome and was genotyped by sequencing of the VP1 gene. HPeV was detected in 20 of 247 specimens tested, and the detection rate was found to be 8.1%. Seventeen of the 20 strains that tested positive for HPeV were sequenced successfully the VP1 gene. The majority of the HPeV strains (n = 15) could be identified as HPeV1, and the remaining 2 strains could be typed as HPeV3. By phylogenetic and identical matrix analyses of HPeV VP1 sequences, HPeV1 should be divided into two lineages, and all of the Japanese studied HPeV1 strains belong to the lineage 2 accordingly. This is the first report of the circulation of HPeV, especially HPeV1 in Japan.

• A single-tube multiplex PCR for rapid detection in feces of 10 viruses causing diarrhea.

  Authors: Pattara Khamrin, Makiko Okame, Aksara Thongprachum, Nattika Nantachit, Shuichi Nishimura, Shoko Okitsu, Niwat Maneekarn, Hiroshi Ushijima

  Journal of virological methods. 02/2011; 173(2):390-3.

  A novel multiplex polymerase chain reaction assay was developed to identify 10 viruses in a single tube. The assay was targeted to detect group A and C rotaviruses, adenovirus, norovirus GI,A novel multiplex polymerase chain reaction assay was developed to identify 10 viruses in a single tube. The assay was targeted to detect group A and C rotaviruses, adenovirus, norovirus GI, norovirus GII, sapovirus, astrovirus, Aichi virus, parechovirus, and enterovirus. A total of 235 stool samples were collected from infants and children with acute gastroenteritis in Kyoto, Japan, from 2008 to 2009, then tested by this novel multiplex PCR and compared with a multiplex PCR described previously, which used 3 primer sets. The novel multiplex PCR could detect the targeted viruses in 111 of the 235 (47.2%) stool samples. Of these, 9 out of 10 types of viruses were identified, including group A rotavirus, norovirus GII, enterovirus, sapovirus, adenovirus, parechovirus, group C rotavirus, astrovirus, and norovirus GI. In contrast, the multiplex PCR that used 3 sets of primers could detect the targeted viruses in 109 of the 235 (46.4%) stool samples. Among these, 8 types of viruses were identified, including group A rotavirus, norovirus GII, enterovirus, adenovirus, parechovirus, group C rotavirus, sapovirus, and astrovirus. The results suggested that the new multiplex PCR is useful as a rapid and cost effective diagnostic tool for the detection of major pathogenic viruses causing diarrhea.

• Novel recombinant sapovirus in Bangladesh.

  Authors: Shuvra Kanti Dey, Masashi Mizuguchi, Shoko Okitsu, Hiroshi Ushijima

  Clinical laboratory. 01/2011; 57(1-2):91-4.

  Recombination of RNA viruses plays an important part in molecular epidemiological study, virus evolution, vaccine design, and viral control programs. Sapovirus, a member of the family Caliciviridae,Recombination of RNA viruses plays an important part in molecular epidemiological study, virus evolution, vaccine design, and viral control programs. Sapovirus, a member of the family Caliciviridae, is one of the major causative agents of viral gastroenteritis affecting all age groups. Sapovirus capsid and polymerase regions were amplified by PCR using specific primers. PCR products were sequenced directly and sequence analysis was performed using CLUSTAL X, SimPlot, and MEGA 4 software package. Based on the genetic analysis, a novel, naturally occurring recombinant sapovirus strain was identified in Bangladesh. Breakpoint analysis of the recombinant sapovirus showed that the recombination site was at the open reading frame ORF1/ORF2 overlap. We described the genetic characterization of a novel, naturally occurring recombinant sapovirus and provided the first evidence of recombination in sapovirus in Bangladesh.

• Seroprevalence of rubella and measles antibodies in Lao PDR.

  Authors: Manilay Phengxay, Yuko Hayakawa, Tung Gia Phan, Kumi Uneno-Yamamoto, Keiko Tanaka-Taya, Phengta Vongphrachanh, Katsuhiro Komase, Hiroshi Ushijima

  Clinical laboratory. 01/2011; 57(3-4):237-44.

  Nowadays, measles and rubella epidemics still occur worldwide, particularly in developing countries. The present study investigated seroprevalence of rubella and measles IgG antibodies by using aNowadays, measles and rubella epidemics still occur worldwide, particularly in developing countries. The present study investigated seroprevalence of rubella and measles IgG antibodies by using a comercial enzyme immunoassay. The findings helped to understand the current status of these infections in school children in Vientiane Capital, Lao PDR. The seropositive rate for measles antibody was 97.6%. Most importantly, more than half of 411 healthy children (56.4%) were rubella seronegative and susceptible to acquired rubella infection. The results highlight a high proportion of rubella-seronegative children, suggesting that they are susceptible to rubella and face the risk of congenital rubella syndrome. It was the first study to report on seroprevalence of rubella in Vientiane Capital, Lao PDR.

• Viral molecular characterization of the first congenital rubella syndrome case in Vietnam.

  Authors: Dinh Nguyen Tran, Minh Phuc Vu, Manh Tuan Ha, Tran Phuong Linh Giang, Katsuhiro Komase, Masashi Mizuguchi, Hiroshi Ushijima

  Clinical laboratory. 01/2011; 57(5-6):397-401.

  Rubella virus (RV) infection during the first trimester of pregnancy often leads to severe birth defects known as congenital rubella syndrome (CRS). A premature newborn male was clinically diagnosedRubella virus (RV) infection during the first trimester of pregnancy often leads to severe birth defects known as congenital rubella syndrome (CRS). A premature newborn male was clinically diagnosed as CRS with cataracts, congenital heart defects, microcephaly, hepatosplenomegaly, and thrombocytopenia. The infection was confirmed serologically and molecularly. The RV was characterized and clustered with the 2B genotype. The integrated description of clinical features, serological and molecular data provide a baseline for diagnosis and control of rubella and CRS in Vietnam. This is the first report of molecular investigation of wildtype RV strain in Vietnam, thus contributing to the documentation of RV's worldwide distribution.