We report here a persistent form of Coxiella burnetii infection. There have been no prospective surveys of chronic C. burnetii infection reported in Japan. Until recently, it was not possible to distinguish between previous and current infection with serological tests for antibody to C. burnetii .The nested PCR method, however, allows us to appreciate the current infection by detecting C. burnetii DNA with high sensitivity. Inoculation method using an A/J mouse was performed to confirm the viability of C. burnetii.
To obtain an approximation of the prevalence of C. burnetii infection in the general population, we evaluated a random sample of patients with symptoms of continuous low-grade fever for one month or more. Analysis of 54 subjects with protracted debility and fatigue symptoms identified 13 subjects as carriers of C. burnetii (24.1%). There were no significant differences in age. C-reactive protein levels (0.69±1.19 mg/dl), white blood cell counts (6, 089±2, 189/μl), eosinophil (3.4±3.6%) between the patients with C. burnetii infection and infection-free subjects. All thirteen patients had experienced protracted low-grade fever (up to 37.5°C) for four months to seven years (30.5±27.7 months). Transthoracic echocardiography showed no evidence of endocarditis, or echosonography revealed no abnormal findings in the liver or kidneys. Although domestic animals constitute an important reservoir of C. burnetii, only two of the positive subjects had direct contact with them and none of the positive subjects were occupationally exposed to farm animals or common sources of infection. None had a history of hospitalizations for pneumonia or hepatic disease. Interestingly, five of the thirteen patients had a history of consulting a psychiatrist, and furthermore, one had a history of several admissions in a psychiatric hospital due to chronic fatigue symptoms. Ten of the patients had a high IgE titer (>295IU/ml), which shows a higher prevalence than in patients without C. burnetii (76.9%: 22.0%, P=0.001). Four of them had markedly elevated IgE levels, in excess of 2, 000IU/ml. The mean value of IgE was higher in the patients with C. burnetii infection than in infection-free subjects (1, 388±1, 706: 533±913IU/ml, p<0.045). Two subjects were rheumatoid factor positive and another three had autoimmune thyroiditis. Twelve of the 13 subjects provided written informed consent for treatment with minocycline (200mg/day). One month later, all subjects became asymptomatic and apyretic (37.1±0.43°C to 36.7±0.56°C p<0.025), and nested PCR did not identify C. burnetii DNA in serum samples. It should be noted that persistent symptoms including low-grade fever were observed for two weeks after the start of medication. Furthermore, three patients had persistent symptoms, and DNA detection by the nested PCR method became positive in all three patients within a few months.

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Diarrheagenic Escherichia coli are differentiated from non-pathogenic members with enterotoxin production, enteroinvasiveness and serotyping. However, the serotypic members are rarely sufficient to reliably identify a strain as diarrheagenic on E. coil. Recently, there are many definite articles which the adhesive E. coli strain against intestinal epithelial cells is enterovirulent.
In this study, 1, 748 E. coil isolates of diarrheagenic and non-diarrheagenic categories which belonged to EHEC, ETEC, EIEC EPEC and non-EPEC were examinated by PCR method for the presence of eaeA, aggR and bfpA regarding adherence factor genes, and astA of EASTl. The strains examined were recognized to variable carrying geno-patterns, and a large number of EHEC, EPEC and non-EPEC had carried either eaeA or aggR genes. In EHEC isolates, a carrying pattern with the most high frequency was only eaeA, and this type was recognized in the isolates of serotype O157, O26 and 0111. EPEC and non-EPEC isolates were recognized eaeA or aggR which harboring with astA or not. Of 508 EPEC isolates from human, a total of 137 isolates (27.0%) carried aggR, and a total of 74 isolates (14.6%) had eaeA, while of the 91 isolates from non-human were recognized aggR and eaeA with 2.2 % (2isolates) and 12.1% (11 isolates), respectively. Also, of 266 non-EPEC isolates from human, a total of 16 isolates (6.0%) carried aggR, and a total of 58 isolates (21.8%) had eaeA. On the other hand, 22 (7.0%) of 316 isolates examined from non-human had eaeA, however no isolate had aggR. Thirteen isolates of EIEC and 218 ETEC isolates were screened, and only 6 ETEC isolates had either eaeA or aggR. The astA gene was recognized in the isolates of all categories, and ETEC strains had more frequently. The bfpA gene was recognized with more frequently in a serotype O157: H45, which is obtained from human with diarrhea, however, this strain was not recognized a member of the EPEC serotype.
There is no diagnostic system for the strain of E. coil that cause diarrheal diseases, therefore more laboratories are unable to identify them. The authors had confirmed which PCR tequnique is a useful simple and rapid method for the detection of adherence factor genes on E. coil strains. From the these results, we showed a differentiation method using PCR technique which have relation with adherence factor, enterotoxin-production and invasiveness, and we firmly believe that application of the procedure is a reasonable and useful method for the identification of diarrheagenic E. coil.

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We evaluated the usefulness of a multiplex-PCR method for differentiation of Entamoeba histolytica and Entamoeba dispar, which are morphologically indistinguishable species. Cultured trophozoites of E. histolytica HM-1: IMSS and E. dispar SAW were used as the positive control. Seven human fecal samples, from which E. histolytica-like cysts were detected by microscopic examination, and three intestinal protozoan parasites, Cryptosporidium parvum HNJ-1, Giardia intestinalis Portland-1, and Blastocystis hominis Nand II, were used for the evaluation of sensitivity and specificity of the PCR method. The other PCR method, which has been used for the diagnosis of amebic infections in Japan, was also performed by using the same samples for the evaluation. In comparison with the conventional PCR method, the multiplex-PCR showed 1) higher sensitivity, 2) the size of diagnostic fragments of PCR products was clearly different in both Entamoeba species, 3) it was possible to perform PCR using a single tube per sample, and then to save the amount of DNA polymerase, 4) no diagnostic amplification products were found in other intestinal protozoan parasites, and 5) E. histolytica specific fragment was amplified in all clinical samples examined. In conclusion, it is considered that the multiplex-PCR method is a useful tool for detection of both Entamoeba species DNA from fecal samples and for the distinction between E. histolytica and E. dispar.

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Faropenem (FRPM) is an only penem antibiotics. Though it has been reported that FRPM had good efficacy (overall efficacy rate: 82.0%) against patients with complicated urinary tract infection, FRPM has not been frequently used for UTI patients. This multicenter clinical study was designed to compare FRPM 300mg 3 times daily to Levofloxacin (LVFX), which is the standard treatment for patients with UTI, 100mg 3 times daily for 7days in the treatment of urinary tract infections in patients with neurogenic bladder and/or benign prostatic hypertrophy.
A total of 60 patients with significant bacteriuria and pyuria were included in this study. Overall efficacy rate (excellent plus moderate) was achieved in 90.6% (29/32) of patients treated with FRPM versus 82.1% (23/28) of those treated with LVFX. The ratios of eliminated bacteriuria and cleared pyuria were 71.9% and 56.3% of patients treated with FRPM, and 64.3% and 75.0% of those treated with LVFX. These data were not significant difference. In conclusion, FRPM 300mg 3 times daily is at least as effective as LVFX 100mg 3 times daily in patients with complicated urinary tract infection.

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Minimum Inhibitory Concentration (MIC) of 4 new-quinolone antibacterial reagents and 4 macrolide antibiotics against periodontopathic bacteria 21 species 24 strains of standard strains including oral related bacteria and clinical isolated (1993-1999) 20 strains of Porphyromonas gingivalis, 7 strains of Actinobacillus actinomycetemcomitans were determined. Minocycline HCl was used as the comparative reagent. Macrolide showed moderate antibacterial activity against standard strains of oral related bacteria except Fusobacterium nucleatum. New-quinolone showed excellent activity against Eikenella corrodens and Actinobacillus actinomycetemcomitans. Concerning to the clinical isolated strains, Clarithromycin effected P.gingivalis (MIC₉₀ 0.1μg/ml) and Levofloxacin and Ciprofloxacin showed high-potency antibacterial activity against clinical isolated Actinobacillus actinomycetemcomitans (MIC₉₀ 0.013-0.025μg/ml).

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We investigated the effectiveness of oseltamivir treatment against influenza virus infection in children. We treated 131 patients (mean age, 5.8±3.6 years) with oseltamivir (4mg/kg/day for 5 days) during the 200-2002 epidemic. All of the patients had been diagnosed with influenza using a rapid diagnosis kit. When treatment was initiated within 48 hours of the onset of fever, 44%of the patients became afebrile (<37.5°C) within one day, and 86%of them recovered within two days. The average duration of fever after the initiation of oseltamivir treatment was 1.7 days. Oseltamivir was equally effective against both influenza type A and type B. No differences in the effectiveness of oseltamivir treatment were observed between young children (<4 years of age) and school-aged children (>6years of age). No obvious side effects were observed.

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We report three dengue fever cases, infected during a group tour to the Philippines. A 58-year old male experienced sudden onset of high fever 5 days after returning to Japan, followed by rash and thrombocytopenia. The other 2 cases experienced similar symptoms. Clinically suspected from the travel history, incubation time and the state of dengue fever epidemic in the Philippines, dengue virus infection was confirmed by the laboratory tests. The incidence and geographical distribution of dengue virus infection have greatly increased in recent years. There have been reports of Japanese travelers who visited dengue endemic countries, infected and developed symptoms after returning home. Dengue virus infection should be included in the differential diagnosis of the patients who develop high fever and rash after returning from tropical areas.

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A 20-year-old male was admitted to our hospital with the chief complaints of high fever and pain around his right hip joint. He had his right knee injured with a slight abrasion three weeks before. The diagnosis of suppurative lymphadenitis of inguen was made, and intravenous cefotiam was started. Despite these treatments his fever continued, general edema and dry cough appeared. Arterial blood gas showed severe hypoxia and chest X ray revealed marked cardiomegaly associated with ground-glass opacity over bilateral lower pulmonary fields. Slight renal insufficiency was also observed. On the fifth hospital day, the culture specimens of both blood and pus from the abrasion on admission yielded Streptococcus pyogenes. His condition was diagnosed as severe group A Streptococcus infection, then antibiotics were switched to intravenous administration of high dose aminobenzyl penicillin and clindamycin in combination with protease inhibitor, urinastatin. After these treatments, his condition improved and he was discharged from the hospital after one month.
Group A Streptococcus may cause uncommon but life-threatening infection such as septicemia. Early recognition of the disease and prompt initiation of appropriate treatment may lead to successful outcome.

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A case of human parvovirus B19 (B19) infection is reported. A 6-year-old previously healthy girl was admitted to our hospital complaining of slight fever and petechial hemorrhage on her neck, trunk and the proximal parts of extremities. On admission, the platelet count was within normal range (180×10³/μl) but white blood cells and reticlulocytes were moderately suppressed (2.4×10³/μl and 1‰, respectively). The purpura disappeared in a week and the blood cell counts fully recovered without any specific treatment. Detection of B19 DNA and anti-B19 IgM antibody in the serum on admission led to the final diagnosis. Since the cellular receptor for B19, the blood group P antigen, is expressed on vascular endothelial cells as well as erythroid progenitor cells, the purpura was considered to be the result of direct vascular injury. She was very unique as she subsequently exhibited papular-purpuric gloves and socks syndrome and erythema infectiosum during follow-up. This case may provide a new insight into the pathogenesis of cutaneous manifestations of B19 infection.

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