In a previous paper, we presented a basic study of an enzyme-linked immunosorbent assay with avidin-biotin system (A-B ELISA) for the detection of serum Candida antigen. In this report, the usefulness of A-B ELISA was evaluated in experimental systemic candidiasis of rabbits and patients with candidiasis. The serum Candida antigen was detected from the first day until death (7 days after inoculation) in a fetal rabbit. However, in non-fetal rabbits, the antigen was detected only on the early stage of infection. In clinical cases, the antigen was detected in 5 patients with candidiasis by the assay. They all were compromised hosts, neonates 2, acute lymphocytic leukemia 2 and one suspected of acute myelofibrosis. Blood samples from the 3 patients grew Candida. In one of them, the antigen had been detected while C. albicans was recovered from her blood. In another case, the antigen was detected persistently even after C. tropicalis disappeared from her blood due to a treatment with antifungal agents. Candida was not recovered from blood samples of the other two. However, we succeeded to detect the antigen from them. Judging from this result in addition to their clinical features and laboratory data, they were suspected of systemic candidiasis. When they were treated with antifungal agents, their clinical symptoms and laboratory findings were improved, and antigen disappeared. These results seem to suggest that A-B ELISA is useful for the early diagnosis of systemic candidiasis.
Three hundred and thirty-seven strains of Vibrio parahaemolyticus, that is, 207 from human feces and 130 from foods, and 22 strains of Vibrio alginolyticus from shellfishes were examined in vitro for their susceptibility to 11 antimicrobial agents; sulfonamide (SA), doxycycline (DOXY), chloramphenicol (CP), kanamycin (KM), erythromycin (EM), polymixin B (PB), ampicillin (ABPC), carbenicillin (CBPC), cephalothin (CET), latamoxef (LMOX) and nalidixic acid (NA). The results can be summerized as follows: 1) Forty-four out of 337 V. parahaemolyticus strains (13.1%) and 20 out of 22 V. alginolyticus strains (90.9%) were drug resistant. 2) As for the resistant strains of V. parahaemolyticus, 4.2% was resistant to ABPC, 3.3% to CBPC, 0.3% to CET, 6.8% to KM and 2.1% to SA, while 86.4% of drug resistant V. alginolyticus strains was resistant to ABPC, 95.5% to CBPC, 18.2% to CET, 0% to KM and 4.5% to SA. 3) Among the one-drug resistant strains of V. parahaemolyticus, those to KM-single were the most predominant (6.2%). 4) None of the examined strains were resistant to DOXY, CP, EM, NA and LMOX.
The value of hydrophobicity test based on salting out was examined by using Escherichia coli isolated from traveller's diarrhea in order to diagnose enterotoxigenic E. coli. In all, 821 E.coli isolates from 180 diarrheal patients were tested and the following results were obtained. 1) Out of 157 strains of positive hydrophobicity test, 137 and 20 strains were enterotoxigenic and non-enterotoxigenic E. coli, respectively. 2) Out of 664 strains of negative hydrophobicity test, 29 and 635 strains were enterotoxigenic and non-enterotoxigenic E.coli, respectively. 3) Among strains of positive hydrophobicity test, 54, 17 and 66 produced only heat-stable enterotoxin (ST), only heat-labile enterotoxin (LT) and both ST and LT, respectively. 4) Screening efficiency for identifying enterotoxigenic E.coli by salting out test was 83% in sensitivity and 97% in specificity. These data suggest that the hydrophobicity test using salting out is a rapid, inexpensive, and simple screeng test for enterotoxigenic E. coli.
The clinical efficacy and safety of HBK were compaired with those of amikacin (AMK) in 230 patients with respiratory infections (pneumonia or lung abscess, chronic respiratory tract infections) by means of a double-blind study at 50 institutions in Japan. Patients over 15 years old with apparent clinical signs and symptoms were administered HBK or AMK intramuscularly b. i. d. for 14 days with a daily dose of 200 mg (pot.) of HBK or 400 mg (pot.) of AMK. The parameters assessed were clinical efficacy, rate of improvement of clinical signs and symptoms, bacteriological response, appearance of side effect, abnormal laboratory findings and clinical usefulness. Clinical efficacy was analysed statistically in 199 patients (100 administered HBK, 99 administered AMK), but 31 patients out of a total of 230 patients were excluded. Side effects were also anlaysed in 213 patients (HBK: 108, AMK: 105) in whom judgement was possible. 1. On the basis of committee judgement, the overall clinical efficacy rate for pneumonia or lungabscess was 63.2% for HBK and 69.4% for AMK. Those for chronic respiratory tract infections were 46.5% for HBK and 60.0% for AMK. No significant difference was observed between the two drug groups. In the evaluation of clinical efficacy by the doctor, the overall efficacy ratesinpneumonia or lung abscess cases were 70.4% for HBK and 61.2% for AMK. Those for chronic respiratory tract infections were 55.8% for HBK and 62.0% for AMK. No significant difference was observed between the two drug groups. 2. With respect to the rate of improvement of clinical signs and symptoms, the HBKgroup was superior to the AMK group in the improvement of sputum volume five days after treatment (p<0.05) in patients with pneumonia or lung abscess. In patients with chronic respiratory tract infections, the AMK group was superior to the HBK group in the improvement of body temperature and CRPseven days after treatment (p<0.05). 3. The bacteriological eradication rate of causative organisms was 38.3% out of 51 patients treated with HBK and 37.5% out of 50 treated with AMK. There was no significant differencebetween the two groups. 4. Some side effects were observed in 10 patients out of 108 in the HBK group, butnone were observed in the AMK group. The HBK group displayed a significantly higher rate of side effects than the AMK group. Abnormal laboratory findings were observed at the rate of 23.4% forHBK and 20.0% for AMK, without significant differences between the two groups. 5. Regarding usefulness as judged by committee members, the respective rate for the HBK group and AMK group were 53.8% and 64.6%. Usefulness, as judged by the doctor in charge, was 63.0% for the HBK group and 63.3% for the AMK group. These assessments showed no significantdifferences between the two groups. From the above results, it was concluded that HBK is as equally useful as AMK for the drug treatment of respiratory infections.
Although there are several tools for serological diagnosis of group A streptococcal infection or its sequelae, the serological reactions are related to the extracellular products of group A streptococcus. Therefore, it is considered worthy to investigate the method of determination of antibodies in human sera to the group-specific polysaccharide (C-polysaccharide, C-poly.) which is one of the somatic antigen of group A streptococcus. Recently, passive hemagglutination test has been utilized on the measurement of anti-C-poly antibody. The present paper describes the fundamental studies on the measurement of the antibody against Cpoly in the immunized rabbit or human sera using the enzyme-liked immunosorbent assay (ELISA) technique. The purified C-poly antigen was extracted from cell walls of group A streptococcus. The C-poly was coupled with poly-L-lysine (PLL) using cyanuric chloride as coupling agent. The C-poly-PLL antigen was coated to microplate wells. The results obtained were as follows: 1. When 0.7μg/well of C-poly-PLL antigen reacted with 100μl of 1: 1, 000 dilution fluid of human sera, the highest titer was shown as O. D. value (410nm) on ELISA test. 2. Absorption test with whole cells of group A, B, C, D, and G streptococcus and Str. pneumoniae, purified group A C-poly and haptenic sugar (GlcNAc) indicated that the antibody specific to Nacetylglucosamine residues of C-poly is detected on ELISA test. 3. The inactivation (at 56°C for 30min) of serum did not affect on the measurement of antibody by ELISA test. 4. There was relatively correlation between anti-C-poly antibody levels of IgM class which were measured by ELISA and anti-C-poly antibody levels which were detected by passive hemagglutination test (r=0.62), but no correlation between the antibody levels of IgG class by ELISA and the antibody levels by passive hemagglutination test. 5. When the antibody titers (IgG class) against the C-poly-PLL in the sera of patients with acute post-streptococcal glomerulonephritis and rheumatic fever were compared with those in the sera of healthy individuals using ELISA test, the titers of the former sera were significantly higher than those of the latter sera (p<0.01). There were no differences, however, in the antibody titer of IgM class to the Cpoly-PLL between sera of patients and healthy individuals. The measurement of the anti-C-polysaccharide antibody by ELISA technique is considered to be a useful tool as one of the serological diagnosis of streptococcal sequelae.
Antibiotic susceptibility, β-lactamase activity and biotype of H. influenzae or H. parainfluenzae isolated from various clinical specimens from July 1982 to January 1984 were investigated. The result were as follows: 1. 231 strains of ABPC sensitive H. influenzae did not produce β-lactamase. 104 strains of H. influenzae were ABPC resistant, 101 of them produced β-lactamase and 3 of them did not produced. 2. 66 strains of β-lactamase producing H. influenzae were also resistant to SBPC, PIPC, MZPC.β-lactamase producing ABPC resistant strains were not resistant to cephems. CTX, CZX, CMX were very effective to β-lactamase producing ABPC resistant strains. 3. 5 strains of H. influenzae, which were resistant to CMZ, CTM by the disk diffusion method, were resistant to ABPC and MICs of CTM, CMZ, CTX, LMOX, AZT were elevated slightly. 4. Nitrocefin disk method and pH disk method could detect the β-lactamase of H. influenzae. 5. Marked differens of biotype distribution was not observed between β-lactamase producing ABPC resistant and β-lactamase non-producing ABPC sensitive H. influenzae. 6. Susceptibility pattern was almost identical, H. parainfluenzae isolated from respiratory tract with H. influenzae. 5 strains of H. parainfluenzae isolated from urinary tract were resistant to CTX, LMOX, AZT, but 2 strains of them produced β-lactamase.
Antibody to delta antigen (anti-delta) was determined in the sera of 321 cases with hepatitis B virus (HBV) infection in Kure, Japan. The following results were obtained. 1) Of 312 cases with HBV infection who were tested for anti-delta, 18 (5.6%) were seropositive. HBV carriers with chronic liver disease had a greater frequency of seropositivity of and-delta than asymptomatic HBV carriers (7.0% vs 0%, p<0.05). 2) In various geographic regions of Kure, the east region had a significantly higher prevalence of anti-delta when compared with all other regions combined (9.8% vs 2.7%, p<0.025). HBV carriers with chronic liver disease in the east region had a greater frequency of seropositivity of and-delta than asymptomatic HBV carriers (15.5% vs 0%, p<0.05).
From 1975 to 1985 in Tokyo and the vicinity, 197 strains of Bordetella pertussis and 16 strains of Bordetella parapertussis isolated from patients of whooping cough. B. parapertussis strains were constituted about 7.5 per cent of the strains isolated from patients. Serotyping of the 197 strains of B. pertussis were carried out by the slide agglutination test using monospecific factor sera. Eleven strains (5.6%) belonged to serotype 1.2.3, 186 strains (94.4%) to 1.3. All the isolates of B. pertussis were highly susceptibel to piperacillin and cefoperazone, and also to erythromycin, minocycline, josamycin, latamoxef, tetracycline, amoxicillin, cefotaxime, ampicillin, and chloramphenicol, and were less susceptible to benzylpenicillin, nalidixic acid, cephalothin, streptomycin, cefatrizine, and sulfamonomethoxazol, and least susceptible to cefaclor and cephalexin. All the isolates of B. parapertussis were highly susceptible to piperacillin, cefoperazone, and minocycline, and also to latamoxef, erythromycin, and tetracycline, and were less susceptible to chloramphenicol, ampicillin, amoxicillin, nalidixic acid, cefotaxime, josamycin, sulfamonomethoxazol, benzylpenicillin, and cephalothin, and least susceptible to cefatrizine, cefaclor, streptomycin, and cephalexin. No drug-resistant strains were found.
The pathogenesis of Mycoplasma pneumoniae (M. pn.) induced pneumonia is poorly understood. In the present study, the cellular analysis in BALF and pathologic studies of the lung in both young and adult hamsters infected with M. pn. were performed to investigate the kinetics of inflammatory and immunologic events in M. pn. induced pneumonia. The results are as follows; 1. The titers of mycoplasmacidal antibody in serum of both hamsters were elevated in all cases. 2. Isolation of M. pn. from trachea and BALF of both hamsters reached its highest point of development on 7th day after inoculation, and the number of organisms isolated from trachea and BALF of young hamsters were as much as those of adult hamsters. 3. Transtracheal inoculation of M. pn. into adult hamsters caused bronchitis, perivasculitis and alveolitis characterized the infiltrations with polymorphonuclear leucocytes (PMNs) against a background of mononuclear cells on 7th day after inoculatin. In cellular analysis in BALF, high population of PMNs in BALF on 7th day and high population of lymphocytes in BALF on 21th day were recognized. 4. In case of young hamsters, there were much less severe lung lesions and there were almost no appearances of PMNs in BALF and lung tissues. These results indicate the importance of cell-mediated reaction in the pathogenesis of M. pn. infection, particularly the infiltration of PMNs appear to play an important role in the early stage of M. pn. induced pneumonia.
We report the first case of meningitis caused by Haemophilus influenzae type Ib resistant to ampicillin and chloramphenicol in Japan. A 1-year-8-month-old girl was admitted to our hospital with 2 days' history of fever elevation and vomiting. On admission the cerebrospinal fluid showed the white cell count of 30, 720/3 mm3, the glucose level of 16 mg/dl, and the pretein count of 177 mg/dl. The Gram stain of direct smears from CSF revealed numerous Gram-negative bacilli. Intravenous therapy was started with both ampicillin 333 mg/kg/day and chloramphenicol 83 mg/kg/day. On the next day culture of the initial CSF yielded heavy growth of H. infl. b. The rapid-β-lactamase test showed positive. Disc diffusion pattern showed resistance to ampicillin and susceptivity to chloramphenicol and latamoxef. Ampicillin and chloramphenicol therapy was changed to intravenous latamoxef (300mg/kg/day) therapy. The minimal inhibitory concentrations (MICs) were determined later by agar dilutions method. The inoculum size was approximately 106/cfu/ml. The result showed ABPC-MICs of 62.5μg/ml and CP-MICs of 12.5μg/ml, and revealed resistant to both ampicillin and chloramphenicol. And also the chloramphenicol acetyltransferase assay was positive. On 8th hospital day Computed Tomography was performed and subdural empyema was found in left frontal area. Burr holes were placed with no complications. She was received latamoxef until the twenty-fifth hospital day. She was dischaged on the twenty-eighth hospital day. She was readmitted at nine weeks after discharge because of the febrile convulsion. But her physical and neurological examinations, including computed tomography and electroencephalogram, were all within normal limits. We conclude that third generation cepalosporines are useful in cases of the meningitis caused by Haemophilus influenzae resistant to ampicillin and chloramphenicol.