We developed a reverse transcription and polymerase chain reaction (RT-PCR) method for detecting astrovirus serotypes 1, 2, 3, 5, 6 and 7 (but not serotype 4). Furthermore, we developed the specific primers for detecting serotypes 1 and 2, the most predominant serotypes in the world. Sensitivity of the first PCR with serotype common primers was about 10 times higher than that of enzyme immunoassay with monoclonal antibody (EIA-MAb). Sensitivity of the second PCR with the serotype-specific primers was even higher. The RT-PCR method was useful for detecting astroviruses from clinical samples, especially serotypes 1 and 2.

Rotavirus (44.7%), adenovirus (5.6%), small round structured viruses such as astrovirus (9.8%) and Norwalk like virus (6.9%) were detected by latex agglutination and reverse transcription polymerase chain reaction amplification from 378 stool samples in an outpatient clinic of Pediatrics in Maizuru area from 1991 to 1994. 70.0% were found from 6 months to 2 years old and 91.0% were detected from January to April. Serotype 1 astrovirus and group 2 Norwalk like virus were mainly found in all serotypes in each virus. Diarrhea, vomiting, nausea, fever, cough and rhinorrhea, and severity of diseases were examined in each case. Fever is significantly found in patients with rotavirus. Sporadic cases with small round structured viruses were recognized in the outpatient clinic of pediatrics.

ObjectiveA novel biocatalyst for Baeyer-Villiger oxidations is necessary for pharmaceutical and chemical industries, so this study aims to find a Baeyer-Villiger monooxygenase (BVMO) and to improve its stability by immobilization. ResultsAcetone, the simplest ketone, was selected as the only carbon source for the screening of microorganisms with a BVMO. A eukaryote, Fusarium sp. NBRC 109816, with a BVMO (FBVMO), was isolated from a soil sample. FBVMO was overexpressed in E. coli and successfully immobilized by the organic-inorganic nanocrystal formation method. The immobilization improved the thermostability of FBVMO. Substrate specificity investigation revealed that both free and immobilized FBVMO were found to show catalytic activities not only for Baeyer-Villiger oxidation of ketones to esters but also for oxidation of sulfides to sulfoxides. Furthermore, a preparative scale reaction using immobilized FBVMO was successfully conducted. ConclusionsFBVMO was discovered from an environmental sample, overexpressed in E. coli, and immobilized by the organic-inorganic nanocrystal formation method. The immobilization successfully improved its thermostability.

TESTPACK ROTAVIRUS(R), a simple 10 min enzyme immunoassay, was compared with reversed passive hemagglutination assay (RPHA) and polyacrylamide gel electrophoresis of virus RNA (RNA-PAGE) for the detection of rotaviral antigens in feces from 104 diarrheal children. The reults of TESTPACK ROTAVIRUS(R) were identical with those of RNA-PAGE. Discordant results of RPHA were further examined by latex agglutination and electron microscopy, and were finally decided to be negative. Pararotaviruses, adenoviruses and small round viruses were negative in the TESTPACK ROTAVIRUS(R) .

ObjectiveA novel biocatalyst for Baeyer–Villiger oxidations is necessary for pharmaceutical and chemical industries, so this study aims to find a Baeyer–Villiger monooxygenase (BVMO) and to improve its stability by immobilization. ResultsAcetone, the simplest ketone, was selected as the only carbon source for the screening of microorganisms with a BVMO. A eukaryote, Fusarium sp. NBRC 109816, with a BVMO ( F BVMO), was isolated from a soil sample. F BVMO was overexpressed in E. coli and successfully immobilized by the organic-inorganic nanocrystal formation method. The immobilization improved the thermostability of F BVMO. Substrate specificity investigation revealed that both free and immobilized F BVMO were found to show catalytic activities not only for Baeyer–Villiger oxidation of ketones to esters but also for oxidation of sulfides to sulfoxides. Furthermore, a preparative scale reaction using immobilized F BVMO was successfully conducted. ConclusionsFBVMO was discovered from an environmental sample, overexpressed in E. coli , and immobilized by the organic-inorganic nanocrystal formation method. The immobilization successfully improved its thermostability.