Three primer pairs, AV244Mon and 82b, AV-15Gr and AV-12Gr, and Beg and End, were used for the detection of standard astroviruses (serotype 1 to serotype 7) in cell culture and astroviruses isolated from 89 diarrheal stool specimens negative for pathogenic bacteria, rotavirus and adenovirus by the reverse transcription-polymerase chain reaction (RT-PCR). All the standard strains and two isolates were detected by primer pairs AV244Mon and 82b, and AV-15Gr and AV-12Gr. All serotypes except serotype 4 were detected by the primer pair Beg and End. The molecular sizes of the RT-PCR products obtained by Beg and End were almost the same among the different serotypes except serotype 6. Sequence analyses of the nucleic acid in the regions between the primer pairs AV244Mon and 82b, and between Beg and End from PCR products of the standard strains and from the isolates of the two stool specimens revealed that each serotype had a unique sequence. These results indicate that the RT-PCR is useful for detecting and genotyping astroviruses.
Recently, the increase in the number of resistant strains of Campylobacter jejuni to fluoroquinolone has been reported in European countries. We also studied antimicrobial susceptibilities of 600 clinical isolates of Campylobacter jejuni isolated during a 6 year period from 1989 through 1994 in four Tokyo Metropolitan Hospitals. The susceptibility to 6 antimicrobial agents, norfloxacin (NFLX), ofloxacin (OFLX), ciprofloxacin (CPFX), nalidixic acid (NA), erythromycin (EM) and tetracycline (TC) were examined. The overall resistant rates were as follows: NFLX, 45 strains (7.5%); OFLX, 45 strains (7.5%); CPFX, 44 strains (7.3%); NA, 62 strains (10.3%); EM, 4 strains (0.6%) and TC, 259 strains (43.2%). The number of resistant strains to fluoroquinolones and NA has increased significantly since 1993 in Japan, but the suscetibility to erythromycin has still remained the same level during the past 6 years. The susceptibility to TC was variable, and MICs gave a bimobal distribution, as pointed out previously. The resistance pattern of NFLX, OFLX, CPFX and NA were observed most frequently in those isolates.
The prevalence of Vibrio cholerae contamination in river water derived from 20 sites of 18 rivers in Kanagawa, Japan, was investigated during a period from July to September, 1987, and from one of the 20 sites in August, 1988 and in February, 1989. V. cholerae non-O1 was found in all samples at concentrations of 0.9->1, 400 MPN/100 ml. Higher amounts of the organism were observed in the samples from estuaries. V. cholerae O1 was detected in samples collected in August, 1988 and in February, 1989 at concentrations of 150 MPN/100 ml and 1.5 MPN/100ml, respectively. From 1989 to 1995, water samples were collected monthly from 10 sites of 10 rivers to detect V. cholerae. V. cholerae O1 and non-O1 were detected in 3.6%(30 of 840) and in 61.1%(513 of 840) in the water samples examined, respectively. Overall, V. cholerae was found in 62.9%(528 of 840). Both types, O1 and non-O1, of organisms were detected in 15 samples. These results indicated that river water was contaminated frequently with V. cholerae non-O1 and sporadically with V. cholerae O1 throughout the year. Only one strain of V. cholerae O1 out of 543 V. cholerae strains was found to be a producer of cholera toxin. During these studies, the selectivity of 3 media for V. cholerae O1 was evaluated, and PMT agar was found to be the best.
The present multicenter study was performed to evaluate the effect of recombinant human granulocyte-colony stimulating factor (rhG-CSF) on combination therapy using aztreonam (AZT) and clindamycin (CLDM) to treat severe infection in neutropenic patients with hematologic diseases. Forty-three neutropenic patients with infections (rhG-CSF group) were treated with AZT (2 g) and CLDM (600 mg) 2-3 times daily as well as rhG-CSF (Lenograstim or Filgrastim: 2-5μ/kg/day). The clinical efficacy of this regimen was compared to that obtained in 44 febrile neutropenic patients, with hematologic diseases, who received only AZT and CLDM in a previous study (historical control group). The overall efficacy rate was 69.8%(30/43) in the rhG-CSF group and 65.9%(29/44) in the historical control group. Although the neutrophil count was significantly increased and C-reactive protein tended to be lower in the rhG-CSF group, the daily maximum body temperature profiles of the 2 groups were nearly the same. These results suggest that rhG-CSF is of little benefit in the treatment of single infectious episodes in neutropenic patients, and that appropriate antibiotic therapy is more important.
We studied the micro-agglutination method (MAT) for the diagnosis of legionellosis. Serum samples were collected from 44 clinically legionellosis suspected patients (17 positive with indirect immunofluorescent antibody technique [IFA] and 27 IFA negatives) and 20 healthy adults (25-30 years old). MAT showed negative results with sera collected from healthy adults and IFA negative patients. 8 out of 17 cass of IFA positive patients showed positive results in MAT. The remaining 9 cases out of 17 were negative in MAT judging from our criteria (1: 256 in single serum or fourfold rise to 1: 128 in pair sera) MAT had good proportion to IFA in samples collected within three weeks after onset of each disease. All sera that became positive in MAT were sampled within four weeks after onset of ach illness. It was noted that MAT was mainly related to IgM-class antibodies. These relationships must be decided by investgating more cases of legionellosis. According to the result of this study, the MAT method was thought to be useful for rapid diagnosis of legionellosis.
To investigate the relationship between serum albumin level and incidence of febrile episodes and mortality in the elderly, we studied 748 patients hospitalized for over one year. The subjects included 123 males and 355 females with a mean age 81.2 years. The average serum albumin level was 3.79 g/dl and levels of serum albumin decreased with advancing age. The incidence of febrile episodes was 1.8 per year in patients with serum albumin levels over 4.1 g/dl, increasing with decline of serum albumin levels. The incidence of febrile episodes was 5.3 per year in patients with serum albumin levels under 3.0 g/dl. Patients with serum albumin levels under 3.0 g/dl displayed a high incidence of febrile episodes irrespective of age. Age adjusted in-hospital mortality was 40.4% during the observed period in patients with serum albumin levels under 3.0g/dl, significantly higher than that of the patients with serum albumin levels over 3.1 g/dl. Relative risk of febrile episode and mortality calculated using the patients with serum albumin levels over 4.1 g/dl as a control was 2.9 and 2.1, respectively, in the patients with serum albumin levels under 3.0 g/dl. These results indicate that serum albumin level is a simple, but strong, predictor of susceptibility of febrile episode and death. Patients with serum albumin levels under 3.0 g/dl may constitute a high risk group for febrile episode and death.
The hemolysin of Aeromonas sobria is one of the important virulence factors in this organism. Rapid detection and identification test for A. sobria is important for early and specific diagnosis of this infectious disease. We evaluated the Polymerase chain reaction (PCR) for the rapid detection of A. sobria. Two pairs of synthetic oligonucleotide primers (ASA1-s and a; AerAAS-s and a) were used in PCR technique to detect the different hemolysin genes (ASA1 and aerAAS) in A. sobria. The PCR identified 91% of ASAl-positive and 23.4% of aerAAS-positive strains in β-hemolytic A. sobria. Other species of Aeromonas, Plesiomonas shigelloides, Vibrio cholerae 01 and V. parahaemolyticus tested were negative in the PCR with two pairs of primers. The PCR technique for detection of two hemolysin genes suggested the possibility of application of this method for detection of A. sobria in A. sobria-associated infections.
Thirty-three stocks of Pseudomonas aeruginosa which is an important etiologic agent of opportunistic infecitons were clinically isolated. The pillin structural gene pilA of the stocks were amplified by polymerase chain reaction (PCR) and classified into 3 groups; 2000 bp (16 stocks; 48.5%), 1300 bp (11 stocks; 33.3%), 550 bp (6 stocks; 18.2%). The adhesiveness of the stocks to cultured human lung cancer origin calu-1 was also determined, their adhesion rate per cell were 39.2%, 24.8%, 22.1% in average respectively. Thus clinically most common 2000 pb group is remarkably easier to adhere to calu-1. Serotypes of the strains were examined to reveal the difference of the distribution that F, G, I types were dominant in 2000 pb group, but E types were major in 1300 bp and 550 bp groups. These data sugest that the gene arrangement of pilA influences adhesiveness to cultured cell and antigenicity of bacteria.
We experienced a SLE patient with TSS after delivery. A 32-year-old SLE patient was transferred to our division due to fever, diarrhea, erosive rash, pericardial effusion, myalgia, lowblood pressure, thorombocytopenia and hypoproteinemia which appeared two days after transvaginal delivery. At the time of admission, we considered these symptoms as the exacerbation of SLE, and treatment with high doses of steroid was started. It was when TSST-1-producing-MRSA was cultured from the vagina and uterus that TSS was suspected. 2 g/day of vancomycin was administered and her symptoms improved. As observed in this case, it is important to consider TSS as one of the complications seen with SLE patients after delivery.
We investigated the possible presence of DNA specific for Aspergillus species in seurm samples of two patients who were strongly suspected for invasive pulmonary aspergillosis (IPA) by a nested polymerase chain reaction (PCR) method. Both patients were diagnosed as having acute myelogenous leukemia and treated with induction chemotherapy. During chemotherapy-induced granulocytopenia, they complained of high fever, and the chest X-rays indicated infiltration shadows in their lungs. They were treated with antibiotics intravenously, but no clinical improvement was observed. As the results of the nested PCR were positive at the acute stage of infection, amphotericin B i. v. and granulocyte colony stimulating factor s.c. administrations were started in both cases. In case 1, the infectious disease improved and the nested PCR results turned negative after treatment. In Case 2, in spite of the progression of the disease, the nested PCR results turned negative during treatment. Although we consider this method very useful for the diagnosis of IPA, further prospective evaluation with a large clinical population sample is required.