The basic studies of detection and quantification of soluble antigen of legionella from urine samples infected with legionella by ELISA were performed. Polyclonal antilegionella antibody was used as the first antibody and alkaline-phosphatase conjugated monoclonal antibody was used as the second antibody in this sandwich assay. Heat solube antigen of L. pneumophila serogroupl (106 CFU/ml) was detectable by conventional ELISA. Heat soluble antigen of L. pneumophila serogroupl (106 CFU/ml) was detectable by the fluorescent ELISA. The sensitivity of fluorescent ELISA method was better than that of the conventional ELISA. Soluble antigen in rabbit urine infected with L. pneumophila was detected from six hours after inoculation of bacteria, and the antigen titer which was equivalent to 106 CFU of heat treated L. pneumophila reached to the maximum at one day after inoculation. The detection of soluble antigen in the urine sample by ELISA was a rapid method and was very useful for the early clinical diagnoses of Leginella disease.
Recent researches on Rickettsia tsutsugamushi isolated from patients of scrub typhus in Japan demonstrated the existence of newly discovered serotypes, such as Shimokoshi, Kawasaki, and Kuroki, which are distinguished serologically from the prototype strains of Gilliam, Karp, and Kato. Additionally, recent observation suggests that, whereas Gilliam and Karp types are dominant in Honshu Island, almost all isolates in Kyushu Island are Kawasaki and Kuroki types. To know whether these serotypes of rickettsiae show identical or different susceptibility to chemotherapeutic agents, MIC values of 6 antibiotics were determined against 3 prototype strains and 15 isolates, such as 3, 2, and 1 isolates of Gilliam, Karp and Shimokoshi types, respectively, isolated in Niigata Prefecture, and 4 and 5 isolates of Kawasaki and Kuroki types, rescpectively, isolated in Miyazaki and Nagasaki Prefectures. The reuslts indicated that all strains tested showed MIC values within 0.021-0.391 μg/ml against tetracycline, minocycline, dimethylchlortetracycline, and leukomycin, and 0.39-6.25 μg/ml against chloramphenicol, whereas no inhibitory effect was observed with aminobenzylpenicillin even at the concentration of 100 μg/ml. Thus, significant differences are not demonstrated in susceptibility to these antibiotics among the isolates and prototype strains of R. tsutsugamushi.
A total of 385 strains of Pseudomonas aeruginosa isolated from patients in Gunma University Hospital was examined for the susceptibility to piperacillin (PIPC), cefsulodin (CFS), ceftazidime (CAZ), imipenem (IPM), aztreonam (AZT), gentamicin (GM), amikacin (AMK), norfloxacin (NALX). Phage type was determined using a set of 12 phages on 90 strains resistant to either drug, and on 37 strains sensitive to above mentioned drugs tested. The results are as follows: 1) Most of GM, AZT and CFS resistant strains were nontypable, and classified as Hh8. 2) This type was specially predominant within the GM resistant strains. 3) All the strains showing serotype (I) and phage type (Hh8) were resistant to both GM and PIPC. It can be concluded that these results were responsible for nosocomial infection.
In order to analyze the serological characteristics of Leptospira interrogans serogroup Icterohaemorrhagiae, monoclonal antibodeies were produced against the serovar smithi Smith strain and the serovar naam Naam strain by cell fusion technology and their properties were examined. Initially, 18 anti-Smith monoclonal antibodies (SMIMA) and 4 anti-Naam monoclonal antibodies (NAAMA) were made, and their serological properties examined by microagglutination test (MAT). Consequently, SMIMA-1 and NAAMA-1, reacting only with homologous strains, were made. Some antibodies recognized common antigen (s) widely distributed among the serogroup Icterohaemorrhagiae except for serovars weaveri, sarmin and tonkini; other antibodies showed different reaction patterns. Next using these monoclonal antibodies, the serological relationships existing among the leptospira serovars of the serogroup Icterohaemorrhagiae were examined. These were divided into three subgroups as follows (1) Serovars monymusk, birkini, mankarso, icterohaemorrhagiae, copenhageni, ndambari, ndahambukuje and budapest, which showed cross-reactivities with the serovar smithi, immunogen leptospira. (2) Serovars weaveri, sarmin and tonkini, which showed no crossreactivities with the serovar smithi. (3) Serovars gem, bogvere, mwogolo, naam and dakota, which showed intermediate characteristics. Also, in subgroup 1, serovars smithi, monymusk, birkini, mankarso, icterohaemorrhagiae and the copenhageni Shiromizu strain showed similar reaction patterns. Serovars copenhageni M20 and Shibaura strains, ndambari, ndahambukuje and budapest showed similar reaction patterns. Serovars monymusk and birkini showed strong cross-reactivities with serovar naam, while serovar icterohaemorrhagiae showed weak cross-reactivity. The antigenic determinants of leptospires were analyzed by the immunoblotting technique using monoclonal antibodies, and were found to react with 18 monoclonal antibodies within a range of molecular mass of approximately 23 kD irrespective of their serological characteristics. The 23-kD molecular mass band was not stained with Coomassie blue but did show a reaction upon silver staining. The band was heat-and proteinase K-resistant, but it was degraded upon treatment with sodium metaperiodate. Accordingly, the band seemed to be a lipopolysaccharide (LPS).
In recent years, there has been an increase of deep mycoses in the patients with a compromised host. We examined clinicopathologically deep mycoses out of 182 autopsied cases experienced in our department from 1981 to 1987, and results obtained are as follows: The incidence of pulmonary mycoses was 23.1% and it was higher in hematologic malignancies (42.3%) than solid ones (8.9%). The common mycoses were aspergillosis and candidiasis which were observed in 18 and 15 cases respectively, followed by mixed mycosis in 6, cryptococcosis in 2 and mucormycosis in one. Most of the patients received large doses of antimicrobial drugs, anticancer drugs and corticosteroids, and showed marked granulocytopenia and lymphocytopenia. It is suggested that the host defence mechanism against deep mycoses chiefly depend on granulocyte counts and function, and cellular immunity.
Five factor sera were prepared with strains of Acinetobacter calcoaceticus subspecies anitratus isolated from clinical specimens. With soft-agar technique using their factor sera, 281 (92.1%) out of 305 strains of Ac. anitratus were typable, 63 (22.4%) out of 281 strains reacted with single factor serum and the remaining strains (77.6%) reacted with polyvalent factor sera. Among them, strains reacted with factor sera of the types Ac-80, Ac-132 and Ac-147 reacted 46.1%. Next, higher frequencies of the strains reacting with the factor sera were in order of Ac-80 and Ac-132, Ac-80, Ac-132, Ac-147 and Ac-156, respectively. As the source of clinical specimens excluding blood and bile, the type Ac-80 and Ac-132 were involved in more than 60% of the strains. On the contrary, strains isolated from blood and bile tended to be non-typable by this technique.
Efficacy, sefety and usefulness of CS-807, a new oral cephem, in the treatment of bacterial penumonia, were evaluated by the double blind comparative method using cefaclor (CCL) as a reference drug. Patients with bacterial pneumonia were treated by oral administration of either 100 mg of CS-807 twice a day or of 500 mg of CCL three times a day for 14 days. The following results were obtained. 1) Out of a total of 186 patients, 144 patients (75 with CS-807: 69 with CCL) were used for comparative evaluation of clinical efficacy. Out of the 144 patients (71 with CS-807: 67 with CCL) had bacterial pneumonia and lung abscess (bacterial pneumonia group) and 6 (4 with CS-807: 2 with CCL) had mycoplasmal pneumonia and primary atypical pneumonia. There was no significant difference in characteristics of the patients between the two treatment groups except that there were a significant number of patients with chest pain in the CCL group than those in the CS-807 group. 2) The clinical efficacy rate judged by committee members in the total cases was 86.7%(65/75) for the CS-807 group and 81.2%(56/69) for the CCL group, respectively. As for the bacterial pneumonia group, the rate was 85.9%(61/71) for the CS-807 group and 82.1%(55/67) for the CCL group, respectively. In both analyses, no significant difference was found between the two grups. According to the judgement by the doctors in charge, the clinical efficacy rate in the total cases was 86.7%(65/75) for the CS-807 group and 83.8%(57/68) for the CCL group, respectively. As for the bacterial pneumonia group, the rate was 85.9%(61/71) for the CS-807 group and 84.8%(56/66) for the CCL group, respectively. There was no significant difference between the two treatment groups in both analyses. 3) As for the bacteriological effectiveness in the bacterial pneumonia group, the eradication rate was 100%(18/18) for the CS-807 group and 85.7%(24/28) for the CCL group, respectively. There was no significant difference between the two treatment groups. 4) The incidence of side effects was 0%(0/89) for the CS-807 group and 3.5%(3/85) for the CCL group, respectively. The incidence of abnormalities in laboratory findings was noted in 19 of 80 cases (23.7%) for the CS-807 group and 21 of 78 cases (26.9%) for the CCL group, respectively. In both analyses, there was no significant difference between the two groups. 5) The utility rate judged by committee members in the total cases was 86.7%(65/75) for the CS-807 group and 77.1%(54/70) for the CCL group, respectively. As for the bacterial pneumonia group, the rate was 85.9%(61/71) for the CS-807 group and 79.1%(53/67) for the CCL group, respectively. In both analyses, no significant difference was found between the two groups. According to the judgement by doctors in charge, the utility rate in the total cases was 86.7%(65/75) for the CS-807 group and 76.8%(53/69), respectively. As for the bacterial pneumonia group, the rate was 85.9%(61/71) for the CS-807 group and 78.8%(52/66) in the CCL group, respectively. There was no significant difference between the two treatment groups in both analyses. As described above, the clinical efficacy and safety of CS-807 at a daily dose of 200 mg were equal to those of CCL at a daily dose of 1500 mg. From these results, it is concluded that CS-807 is an oral antibiotic which is very useful in the treatment of bacterial pneumonia.
A total of 1, 888 patients attending to the obstetrical and gynecological outpatient clinic with various complains were attempted to be diagnosed Chlamydia trachomatis (C. trachomatis) infections by EIA Chlamydiazyme. Furthermore, the serum IgG and IgA titers were measured by ELISA and IFA in 30 Chlamydiazyme and 30 Micro Trak positive cases. 8 cases which were Chlamydiazyme positive and ELISA negative were confirmed by isolation for C. trachomatis using cycloheximide treated HeLa 229 cells. 1) Positive rate in Chlamydiazyme was 9.5% in pregnant women hoping a delivery, 24.2% in prostitutes working in massarge parlor so-called soap-land, 23.1% in students including high school girls. Positive rate in Chlamydiazyme showed fairly high level than the result in Micro Trak reported by us already. 2) Micro Trak positive cases revealed all positive (16 cases) in ELISA, but Chlamydiazyme was 73.3% in same method. 3) IgA titer over ten times in IFA was 66.7% in Micro Trak positive cases and 53.3% in Chlaamydiazyme positive cases. 4) 8 cases showed positive in Chlamydiazyme and negative in ELISA were completely all negative in isolation. In addition, these cases showed less than 80 times in IFA (IgG) and under 5 times in IFA (IgA). From these results, we concluded that Chlamydiazyme is a good for the screening of C. trachomatis infection in uterine cervix, but for the final diagnosis should be performed by directimmunofluorescent test as well as Micro Trak. Furthermore, an elevation of sensitivity on the immunofluorescent test is expected, hereafter.
A 20 year old woman was admitted to our hospital because of fatigue and persistent low grade fever lasting for a 8 month period. Physical examination on admission revealed neither elevation of temperature, lymphadenopathy nor jaundice. Laboratory studies showed increased leukocyte counts with absolute lymphocytosis including atypical lymphocytes, an elevation of GOT, GPT and LDH levels in the serum, a negative heterophil agglutination test and mild hepato-splenomegaly on the radiological scintigram. Anti-cytomegalovirus antibody of IgM class (ELISA) was markedly elevated, while antibodies against hepatitis associated antigen and EB-virus were negative. Histological findings on liver biopsy showed sinusoidal infiltration of mononuclear cells, however, typical inclusions of cytomegalovirus were not identified. Cytomegalovirus was isolated from urine specimens.
A 33-year-old male was sent to our clinic for vomiting, shock, and a skin rash. Descrete pink papules became larger and purpuric within 24 hours, some of which had a central hemorrhage. Physical and laboratory examinations revealed meningitis, herpes labialis, septic shock, DIC, adrenocortical insufficiency, and cardiac failure with evidences of myocardial infarction. N.meningitidis, serogroup B and herpes simplex virus type 1 were identified in the cultured blood and exudates of the swollen lips on the second hospital day, respectively. The patient recovered with medications of ABPC (6-8g/day), heparin, FOY, antithrombin III, and with intensive care for cardiac-septic shock. Hemolytic activities of the patients's compliment were less than 12/CHSO during the course. Screening for each component of the compliments and the reconstruction test of the hemolytic activity with addition of purified seventh component of compliment (C7) disclosed that this patient is a congenital deficiency of C7.