A retrospective clinical study of 64 cases with bacterial meningitus beyond the neonatal period in the department of pediatrics of St. Marys' Hospital (1985-1995) was conducted. Haemophilus influenzae (H.influenzae) (28 cases, 43.8%) and Streptococcus pneumoniae (S. pneumoniae) (23 cases, 35.9%) were common pathogens. The prognosis was classified into three groups; normal (42 cases, 65.6%), neurological sequelae (17 cases, 26.6%) and death (5 cases, 7.8%).
We analyzed the risk factors associated with their outcome. The body temperature at admission, platelet count, CSF examination (WBC, glucose, GOT, GPT) were prognostic factors. The prognosis of bacterial meningitis caused by S. pneumioniae was worse than those due to H. influenzae (p=0.0347).

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Toxic shock syndrome toxin-1 (TSST-1) and enterotoxins are important virulence factors produced by Staphylococcus aureus. It is reported that these toxins are associated with septic shock and toxic shock syndrome. We investigated the toxin production and coagulase types of 701 MRSA strains isolated in Sasebo City General Hospital between 1994 and 1996 TSST or/and enterotoxins were detected in 67% of all MRSA strains, and thous were detected in 88% of MRSA strains isolated from blood samples. 45% of all MRSA strains produced both TSST-1 and enterotoxin C, and 70% of MRSA strains obtained from blood produced those toxins. Frequency of TSST-1 or/and enterotoxin production by MRSA strains isolated from blood samples was significantly higher than that by MRSA strains isolated from urine and pharynx (p<0.05), and frequency of both TSST-1 and enterotoxin C production by MRSA isolates from blood was significantly higher than that by MRSA strains isolated from pharyngeal sample (p< 0.05).
This study indicated that investigation of virulence factors produced by MRSA might give the useful information on prevention and treatment of MRSA infection.

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The prevalence of hepatitis A virus (HAV) antibody in people has decreased from year to year in Japan. A sequential outbreak occurred in an institution for the mentally handicapped after the onset of symptoms. The anti-HAV IgM was found to be positive in 4 sera examined. The HAV antigen was detected in 3 out of 5 feces using ELISA. An existance of inhibitor in 2 negative specimens against the ELISA was suggested by the recovery test of added antigen. HAV RNA was extracted by CTAB method from feces and detected in 4 our of 5 specimens in PCR amplification and in all of 5 specimens in nested PCR amplification. The sequence of PCR products in the P1/P2 junction of the HAV genome revealed that the virus associated with the outbreak belongs to HAV subgenotype IA.
HAV RNA was detected in ELISA negative specimens and in the specimen from a patient days after the onset of symptoms using PCR amplification by CTAB method. These results indicate that PCR amplification was useful for the early diagnosis of hepatitis A infection.

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Serum anti-Bartonella henselae IgG and IgM antibody titers for the diagnosis of cat scratch disease (CSD) were determined by indirect fluorescence antibody (IFA) tests. B. henselae as antigen were harvested either by cocultivating with Vero cells (cocultivated B. henselae) or by cultivating without them (non-cocultivated B. henselae). Based on the results on 110 healthy adults, cut off values were set at 1: 32 for IgG, and <1: 20 for IgM antibodies. According to thesecriteria, IgG antibody was positive in 2.7% of the 110 adults, while nobody was positive for IgM antibody. The titers did not change depending on the types of antigen used.
On the other hand, IgG antibody titers against cocultivated B. henselae tended to be higher than those against non-cocultivated B. henselae in 33 CSD suspected patients; 75.8% of the patients were anti-B. henselae IgG positive when tested with cocultivated B. henselae as antigen, while only 48.5% of the same patients gave positive results with non-cocultivated B. henselae. Anti-B. henselae IgM antibody was positive in 24.2% of the 33 CSD suspected patients against both types antigen. Vero cells themselves seemed to nonspecifically bind some IgM (but not IgG).
We recommened cocultivated B. henselae as antigen for IgG IFA, and non-cocultivated B. henselae for IgM IFA in the serological tests of CSD.

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Intravenously administered immune globulin therapy has been reported to be an effective treatment for serious patients with verotoxin-producing Escherichia coli infections while an efficacy for VT2 are under discussion. We therefore examined in vitro commercially available immune globulin preparations for the presence of anti-VT neutralizing antibodies in expectation of protecting a patient from serious complications as HUS. We examined 47 lots of-globulin prepared by 5 pharmaceutical companies in Japan. Of them 29 lots were prepared from γ imported blood and 18 lots were from Japanese donors' blood. They were prepared in each company following their own manufactures' process.
Neutralizing activity for VT1 and VT2 were determined by means of cytotoxicity using ACHN (renal adenocarcinoma, human) cells. All lots of γ-globulin preparations from imported blood completely neutralized 125pg/ml VT1 at the concentration of 12.5mg/ml, and no significant difference was found in the manufactures' process. On the other hand γ-globulin preparations from Japanese donors' blood neutralized VT1 five times less than the former. None of all preparations neutralized VT2.
Gammaglobulin preparations were produced from pooled human plasma. To know the difference of neutralizing activities in the source, we examined randomly selected 239 human plasma samples of which 51 were from Japanese donors' blood and 188 were from imported blood. Prevalences of neutralizing activity of VT1 in imported plasma and domestic plasma were10.6% and 2.0%, respectively. The prevalence of neutralizing antibody n these plasma samples reflects the different neutralizing activity of VT1 in γ-globulin preparations prepared from imported blood and Japanese donors' blood.
From these results, by selecting the lot of γ-globulin preparations or the material of plasma with highly neutralizing activity, intravenously administered immune globulin therapy may be effective for VT1. The lack of VT2-neutralizing activity in any γ-globulin preparation promotes us to develop humanized anti-VT2-monoclonal antibody for the prevention of HUS in high risk children.

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In this study we investigated the antifungal susceptibility of 285 strains of Candida albicansisolates at Kinki University Hospital from March 1995 to December 1996. The antifungal agents tested were fluconazole, miconazole, itraconazole, amphotericin B and flucytosine. The susceptibility testing were performed according to the broth microdilution method standardized by National Committee for Clinical Laboratory Standards (M27-T). Most isolates of C. albicansshowed relatively a low MIC value and the MIC90s were calculated at 1μg/ml; fluconazole, 0.125μg/mg; miconazole, 0.06μg/ml; itraconazole, 1μg/ml; amphotericin B, 0.25μg/ml; flucytosine. There was only one strain that showed high resistance against fluconazole and it showed cross-resistance against miconazole and itraconazole. There were two flucytosine resistant strains. The MICs of amphotericin B were tightly clustered and resistant strain were not observed.

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We clinically investigated a total of 288 cases of bacteremia for the past ten years, from January 1986 to December 1995, at the Second Department of Internal Medicine in the jikei University Hospital.
All of the subjects who had a positive reaction to blood culture or catheter tip culture were investigated for their basic disease, complications, and detected bacteria. Malignant tumors, chronic renal failure, diabetes mellitus, and hematologic disease were frequent by noted.
The cases due to primary infection were mainly respiratory organ infection or urinary tract infection, which were 47.8% of the total.
In 31.3% of the total, catheter tip cultures were positive.
Except for catheter related infection, Gram-positive coccus were detected in 40.3%, which was most frequent. Methicillin resistant Staphylococcus aureus (MRSA) were 8.1% and Staphylococcus epidermidis were 11.2%. In catheter related infection, Gram-positive coccus were detected in 59.9%, which was most frequent amongst them, MRSA was 17.2%, S.epidermidis was 16.2%.
The mortality of bacteremia was 12.5%, mainly from hematologic diseases, immunodeficiency due to long term steroid administration etc.
Accordingly, the more the advance of chemotherapy, the better the prognosis of septicemia is. Appearance of catheter related infection was unexpected frequent. Increase of immunocompromised host is thought to be one of the main factors in the outbreak of bacteremia.

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A rapid enzyme immunoassay membrane test, Directigen Flu A (Becton Dickinson, USA), was evaluated by using virus isolates and clinical specimens. The reference laboratory diagnosis was based on the results of virus isolation.
Directigen Flu A was reactive for all subtypes of human influenza A viruses, including reference strains of H1N1, H2N2 and H3N2. Moreover, H5N1 (Hongkong/156/97) was also detected by this kit. No cross reactivity was detected with other respiratory viruses.
Directigen Flu A showed positive reaction with the solution containing influenza A virus of 2.4×10³ pfu/assay.
The rapid test demonstrated 77.9% sensitivity and 98.4% specificity for testing of throat swabs from children with respiratory symptoms. It showed higher sensitivity and specificity (92.1% and 100%) for testing of nasopharyngeal aspirates.
Directigen Flu A should be useful for the rapid diagnosis of influenza A virus infection.

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“Quix^TM” is an immunochromato graphy-based direct detection kit for the E. coli O157 LPS antigen in the patient's stool. The present study was conducted to evaluate the efficacy of the kit for rapid diagnosis of enterohemorrhagic Escherichia coli (EHEC) O157 infection.
Sensitivity of the kit was determined using a pure culture of a clinical isolate of E. coli O157. Analytical sensitivity was found to be 5×10⁵ CFU/ml. When compared with the culture method using fecal samples of 64 patients and with bloody diarrhea, sensitivity and specificity were 95.0%(19/20) and 86.4%(38/44), respectively, and overall agreement to culture method was 89.1%(57/64). One patient was found positive by culture method while negative in the present method, where the sample contained a low number of the cells less than the detection limit. Four of the six patients with a negative result by culture method and positive in the present method, were confirmed E. coli O157 infection by positive IgM antibody response against the E. coli O157 LPS. The discrepancy between the two methods seemed to be attributable to antibiotic adminis adminis-tration. In one patient, Salmonella urbana (O30₁30₂) was detected. The O30₁ antigen of this bacterium is well known to be identical to the E. coli O157 antigen.
When the present method was compared with an ELISA-based E. coli O157 LPS antigen detection kit, sensitivity and specificity were 100%(11/11) and 82.1%(23/28), and overall agreement to ELISA method was 87.2%(34/39).
From these findings, Quix^TM is useful as a rapid diagnostic kit in the primary clinics, outpatient or bedside use. E. coli O157 LPS antigen in patient's fecal samples can be detected in about five minutes with this simple procedure. Early diagnosis using such a simple kit will largely contribute for the early treatment and prevention of severe complications of the E. coli O157 infection.

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Recent experience in Japan has suggested that influenza A encephalopathy may occur more frequently than generally appreciated and may have a grave prognosis. Influenza A encephalopathy has been managed to date only with supportive measures, because the efficacy of anti-viral therapy for encephalopathy has not efficacy of anti-viral therapy for encephalopathyhas not yet been documented. During the period from January 1996 to February 1998, we treated two cases of influenza A encephalopathy with amantadine (6mg/kg/day, p.o., for 7 days). Both became status epileptics and had loss of consciousness within 24hours after the onset of illness. They recovered without sequelae and had no side effects from amantadine treatment. Amantadine may be useful in the treatment of influenza A encephalopathy.

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The patient was born by emergency cesarean section forfetal distress at 35weeks gestation with a weight of 2740g. The early neonatal course was complicated by transient tachypnea and renal failure. He was receiving oxygen and diureticus in incubator for 5days and his condition was very improved on day 5. On day 7 he became lethargy and there was inability to tolerate feeding. Investigation of the cerebrospinal fluid revealed 8, 000 leukocytes/μl. S.marcescens was grown from cultures of both blood and cerebrospinal fluid. Treatment was started with cefotaxime and ampicillin every 6 hour. On day 14 the CT showed a brain abscess located parietooccipitally on the left side and diffuse infarction on the right side. On day 14 and 23 recurrence of increased leukocytes in the cerebrospinal fluid, high values of serum CRP and deteriolation of clinical symptoms were observed. It is thought that the episodes show rupture of the abscess into the lateral ventricle. On day 55 surgical drainage was performed for the hydrocephalus. On day 110 the abscess was not found in the brain CT scan. His psycomotor development 3 years later was equivalent to two years old and he had secondary epilepsy.

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A 76-year-old female was admitted to our hospital because of fever and the right pleural effusion. On the analysis of pleural effusion, the total cell count was 6720/μl with 95% lymphocytes, and ADA was 38.1U/1. The culture of pleural effusion was negative, and the smear and PCR for Mycobacterium were also negative. For examinations, we performed eterography that showed cicatricial strictures of intestine. X-ray examination of the colonated colonoscopoy showed ulcers (circular type), shortening of the colon, Bauhin's valve insufficiency and diverticulum-like deformity. Then, she was diagnosed as intestinal tuberculosis. The smear and PCR of biopsy specimens from the lesion were positive, and antituberculotic therapy was effective. Finally, the culture of pleural effusion for Mycobacterium tuberculosis was positive after 8 weeks. We thought intestinal examnaton may be useful for the diagnosis of tuberculosis, when lymphocyte-rich exudative pleural effusion of unknown etiology is seen.

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