We have established a method of quantitative detection and rapid identification of human adenoviruses (hAdVs). Using LightCycler PCR with a primer set, we were able to amplify 554 bp of the hexon gene from each of 51 prototype strains of hAdVs. The sensitivity of LightCycler PCR was 10 copies of hAdV DNA/reaction. When LightCycler PCR was performed using a set of primers, hAdV was positive for 74.4% (99 of 133) of conjunctivitis patients and for 27.3% (81 of 297) of respiratory infection patients. We also attempted to measure hAdV in the potentially contaminated eye drops used by patients, detecting 5.4 ؋ 10 2 to 1.6 ؋ 10 Human adenoviruses (hAdVs) of the genus Mastadenovirus of the family Adenoviridae infect billions of people worldwide and cause various diseases such as conjunctivitis, respiratory infectious disease, diarrhea in infants and young children, hemorrhagic cystitis, etc. (1,8,33,39,40). Most of these diseases heal naturally, but sometimes the infection may also provoke serious illnesses such as pneumonia caused by AdV type 7 (AdV-7) or epidemic keratoconjunctivitis (EKC) due to . These more serious outcomes occur in all age groups and can possibly trigger highly contagious nosocomial infections (5,6,19,27,42). Therefore, it is important to establish a rapid method of virological diagnosis. Furthermore, hAdV infection in immunosuppressed patients, such as graft recipients and immunodeficient patients including those with AIDS, has been a major problem in recent years and has been lethal in many cases (7,9,10,17,23,40,41). Nosocomial infection is also a serious problem which may require restriction of hospitalization and closing of hospital wards (15). Therefore, it is essential to monitor these viruses, and a rapid method of identifying serotypes is urgently needed.hAdVs were initially grouped into six subgenera (A to F) on the basis of several biochemical and biophysical criteria (1, 39). In 1999, reclassification on the basis of nucleotide and deduced amino acid sequences was approved by the International Committee on Taxonomy of Viruses; under this reclassification, the 51 serotypes of hAdVs in the genus Mastadenovirus were grouped into six species, hAdV-A to hAdV-F (38). Virus isolation followed by a neutralization test has been the standard method of serotyping (39); however, these procedures are complicated and time-consuming, and the standardized antisera are in limited supply. Recent advances in amplifying hAdV genes and decoding nucleotide sequences have allowed us to develop a PCR-restriction fragment length polymorphism method with which we have succeeded in distinguishing 14 hAdVs including AdV-3, -4, -8, -19a, and -37, all of which cause eye infections in humans (31). Furthermore, nucleotide sequence analysis of the partial hexon genes (916 bp) of all 33 prototype strains in hAdV-D and hAdV-E allowed us to construct a database for the phylogeny-based identification of hAdVs from patients with conjunctivitis (34). However, this method requires several overlapping sequences to determine 9...

We isolated 872 strains of mumps virus from naso-pharyngeal secretions in seven different districts of Japan from January 2000 to July 2001. Among them, 57 strains were geno-typed by nucleotide sequencing in part of the hemagglutinin-neuraminidase (HN) and small hydrophobic (SH) protein regions. Four different genotypes (B, G, K, and L) of mumps virus were co-circulating in Japan and the distribution of genotypes varied in geographically different districts. Two new clusters designated as genotypes K and L had more than 7% nucleotide variation in the SH gene. Among the 57 strains, 11 were classified as B, 35 as G, three as K, and eight as L, which was mainly isolated in Tokyo. We also examined 104 stains isolated in a clinic in Mie prefecture from 1993 to 2003. Genotype B was the indigenous strain and genotype K was introduced in 1994. Genotypes B and K co-circulated in the 1990s and were replaced by genotype G in 2000. There was no significant change in neutralizing test antibody titers against genotypes B, G, K, and L using seven post-vaccination sera with Hoshino strain (genotype B) and these four genotypes had a different antigenicity from genotype A. We should continue to watch on mumps virus molecular epidemiology.

Although the effectiveness of oseltamivir against influenza virus infection is well known, there has been no report analyzing the detailed time course of fever following the drug treatment in children. Oseltamivir was prescribed for 4 days to every child with a positive result for rapid immunological test for influenza virus during 2002--2003, 2003--2004, and 2004--2005 epidemics. Only those who were 1-12 years of age and prescribed oseltamivir within 24 hr after the onset of fever were included in the analysis. The numbers of children with type A/H3N2 disease for the three seasons were 64, 77, and 33, and those with type B disease were 102, 4, and 86, for the respective seasons. The period until normalization of temperature was obtained from six-hourly recordings of body temperature. By multiple regression analysis, temperature periods were longer in type B than in type A/H3N2 disease, negatively associated with age, and positively with maximal body temperature (all: P < 0.001). The effectiveness of oseltamivir on body temperature in type B disease was less apparent in the 2004--2005 than in the 2002--2003 season, irrespective of age. No such between-season difference was observed for Type A/H3N2 disease. Frequencies of ineffective cases with biphasic fever (19.6% and 43.0% during 2002--2003 and 2004--2005 seasons) were significantly higher in type B than in type A/H3N disease (12.0% and 11.8%, respectively). The effectiveness of oseltamivir depends on a child's age, maximal body temperature and the virus type. This study confirmed recent reports indicating decreased effectiveness of oseltamivir against type B disease.

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