bluuebunny/crossref_metadata_2025 huggingface dataset:
Production and characterization of agglutinating monoclonal antibodies against predominant antigenic factors for Candida albicans
Authors: Miyakawa Y , Kagaya K , Fukazawa Y , Soe G
Published: January 2020

Two clones, CA4-2 and CA5-4, which produced agglutinating monoclonal immunoglobulin M (IgM) antibodies (MAbs) against mannan antigens of Candida albicans serotype A, were established. The specificity of each MAb was determined by slide agglutination tests for cross-reactivity patterns against the homologous and six other strains of Candida and a strain of Torulopsis: C. albicans serotype B, C. tropicalis, C. guilliermondii, C. krusei, C. parapsilosis, C. pseudotropicalis, and Torulopsis glabrata. The MAb produced by CA4-2 reacted with the homologous, C. tropicalis, and T. glabrata strains, whereas the MAb produced by CA5-4 reacted with the homologous, C. albicans serotype B, and C. tropicalis strains. These results are consistent with results obtained by comparative experiments with several strains of each serotype or species. Specificity of these two MAbs by agglutination was also consistent with the cross-reactivity patterns demonstrated by indirect immunofluorescence staining. The competitive binding experiments by immunofluorescence staining with two MAbs and polyclonal factor sera (PAb factors) 5 and 6 suggested that the MAb from clone CA4-2 did not completely correspond to PAb factor 6 and that the MAb from CA5-4 was distinct from PAb factor 5 in its manner of binding to determinants (the latter was designated 5b), Cross-reactivity patterns, however, furnished evidence that these two MAbs could replace the known PAb factors 6 and 5, respectively, as reagents for aid in the identification of the strains of C. albicans and their serotypes.
Sensitivity and specificity of enzyme-linked immunosorbent assay in the detection of antigen in tuberculous meningitis cerebrospinal fluids
Authors: Kadival G V , Mazarelo T B , Chaparas S D
Published: January 2020

A sandwich enzyme-linked immunosorbent assay was developed for its potential utility in the detection of antigen in the cerebrospinal fluid of patients with tuberculous meningitis. Cerebrospinal fluids examined included those from untreated (group Ia) and treated (group Ib) Mycobacterium tuberculosis meningitis, nonseptic central nervous conditions (group II) such as epilepsy, viral meningitis, and tetany, and nonmycobacterial septic meningitis (group III). The average levels of antigens determined and percent positive specimens, respectively, for each group were (group): Ia, 1.8 micrograms/ml and 75% positive; Ib, 0.37 microgram/ml and 36% positive; II, 0.036 microgram/ml and 100% negative; and III, 0.075 microgram/ml and 100% negative. The system developed employed hyperimmune polyclonal antibody raised against M. tuberculosis and Mycobacterium bovis BCG in burros and rabbits. Cross-reactivity by other mycobacterial species was very low; e.g., 5% for M. kansasii and less than 2% for M. intracellulare, M. avium, M. vaccae, and M. fortuitum. The test shows promise as a specific adjunct for the early diagnosis of tuberculous meningitis.
Persistence of chlamydial antibodies after pelvic inflammatory disease
Authors: Puolakkainen M , Vesterinen E , Purola E , Saikku P , Paavonen J
Published: January 2020

The persistence of chlamydial immunoglobulin G (IgG) antibodies and long-term sequelae of pelvic inflammatory disease (PID) were studied in 70 women who had been treated for PID 3 to 6 years previously. Fifty-one women had had PID associated with Chlamydia trachomatis infection (Chlamydia group), and 19 women had had PID not associated with C. trachomatis (non-Chlamydia group). Chlamydial IgG antibodies, as determined by the indirect immunofluorescence test with inclusions of C. trachomatis L2 as antigens, persisted at stable levels in 43% of the women for up to 6 years; 43% of the women showed a decrease in IgG titer, and 13% showed an increase. IgA antibody levels in serum correlated with IgG antibody levels in serum and with the presence of cervical IgA antibodies. Both serum antibodies and cervical IgA antibodies were more often found in the Chlamydia group. Forty-two percent of the women were infertile. Every fifth subsequent pregnancy was ectopic. The presence of cervical IgA antibodies might protect the women from tubal damage.
Nonweekend schedule for BACTEC drug susceptibility testing of Mycobacterium tuberculosis
Authors: Hawkins J E
Published: January 2020

Determination of the drug susceptibility of Mycobacterium tuberculosis by conventional methods using an agar-based medium may take 3 weeks or more to complete. On the other hand, results on positive cultures are generally available in 4 to 7 days with the radiometric (BACTEC, Johnston Laboratories, Towson, Md.) procedure. One disadvantage to the latter is the requirement to determine the quantity of 14CO2 in each test vial on a daily basis from the day of inoculation. Growth index readings often must be made over weekends, adding to the work load of clinical laboratories during periods of reduced staff or necessitating compensatory pay or time. Susceptibility tests with streptomycin, isoniazid, ethambutol, and rifampin against 104 M. tuberculosis strains were performed by the submerged disk method, the recommended BACTEC method with daily growth index readings, and the radiometric procedure with readings delayed for 2 days after inoculation. Criteria for interpretation of "delayed" tests were established. Drug concentrations tested included some modifications of those available commercially. Overall agreement for the four drugs by the three methods was greater than 90%. We conclude that under our test conditions a schedule of inoculation of radiometric test vials on Friday with growth index readings commencing on Monday gives susceptibility results that correlate well with the daily BACTEC method and with a conventional 7H10 agar method.
Immunoglobulin and specific antibody responses to Rhodococcus (Corynebacterium) equi infection in foals as measured by enzyme-linked immunosorbent assay
Authors: Takai S , Kawazu S , Tsubaki S
Published: January 2020

Humoral immune response to intestinal Rhodococcus (Corynebacterium) equi in horses was studied by enzyme-linked immunosorbent assay. Anti-R. equi immunoglobulin M (IgM), IgG, and IgA antibodies were demonstrated in the healthy horse population. Adult horse levels of anti-R. equi IgM and IgG antibodies were reached by 5 to 9 weeks of age in two healthy newborn foals. R. equi was recovered from the foals in the range of 10(3) to 10(4) per g of intestinal contents. A 1-week-old foal was infected with R. equi by mouth daily for 9 weeks. The foal did not show any clinical signs of illness. Anti-R. equi IgM antibody values in the foal increased about 5 to 8 weeks after initial inoculation, similar to the naturally occurring immune response to intestinal R. equi. There were differences among the antibody responses to R. equi in healthy horses, foals with suspected infection, and infected foals. These results suggest that exposure to R. equi is widespread in the horse population and that intestinal R. equi is the most important source of antigenic stimulation for a naturally occurring immune response in horses.
Evaluation of a commercial cytotoxicity assay for detection of Clostridium difficile toxin
Authors: Nachamkin I , Lotz-Nolan L , Skalina D
Published: January 2020

A comparative study was performed to determine the accuracy of Clostridium difficile toxin detection. A commercial cytotoxicity assay (Bartels Immunodiagnostic Supply, Bellevue, Wash.) was compared with conventional microcytotoxicity assays, using Vero and MRC-5 cells. The Bartels system was found to be essentially equivalent to conventional cytotoxicity assays currently being performed for routine C. difficile toxin detection.
First case report from Argentina of fatal septicemia caused by Chromobacterium violaceum
Authors: Kaufman S C , Ceraso D , Schugurensky A
Published: January 2020

A case of Chromobacterium violaceum fatal septicemia is reported. The microorganism was isolated from six blood cultures and two suppurated skin lesions. It is the first such case described in Argentina, and it reinforces the need for prolonged treatment and careful clinical evaluation to ensure complete remission of human infections caused by this bacterium.
Leukocyte survival in cerebrospinal fluid
Authors: Steele R W , Marmer D J , O'Brien M D , Tyson S T , Steele C R
Published: January 2020

Delays in the laboratory examination of cerebrospinal fluid are commonly encountered in clinical medicine. The present studies were designed to evaluate changes in cerebrospinal fluid leukocyte counts relative to time elapsed before analysis. Neutrophil counts decreased most rapidly, being 68 +/- 10% (standard error of the mean) and 50 +/- 12% of initial values at 1 and 2 h, respectively. Lymphocyte and monocyte numbers were not significantly altered until 3 h.
Characterization of Neisseria polysacchareae sp. nov. (Riou, 1983) in previously identified noncapsular strains of Neisseria meningitidis
Authors: Boquete M T , Marcos C , Sáez-Nieto J A
Published: January 2020

A study of 216 noncapsular strains of Neisseria meningitidis isolated from patients and carriers received in the Meningococcus Reference Laboratory between 1978 and 1984 is reported. The characterization of the strains consisted of biochemical tests for the following characteristics used for the differentiation of Neisseria species: oxidase, catalase, and beta-galactosidase activities; sugar degradation; nitrate and nitrite reduction; DNase activity; polysaccharide production with 5% sucrose; aminopeptidase activity; and growth in Thayer-Martin and Catlin media. Of the strains studied, 50 showed characteristics of a new taxon recently described (Neisseria polysacchareae). Characteristics that differentiated these strains from meningococcal isolates were polysaccharide production with 5% sucrose, gamma-glutamylaminopeptidase activity, and a requirement for cysteine or cystine for growth in Catlin medium. All of the N. polysacchareae strains identified were isolated from the nasopharynx of healthy carriers.
An unusual case of cutaneous listeriosis
Authors: Cain D B , McCann V L
Published: January 2020

Listeria monocytogenes was identified as the etiological agent in the cutaneous and febrile illness of a 64-year-old male who acquired the organism as a result of contact with the genital tract of a cow while assisting in the delivery of a stillborn calf.
Sensitivity of different assay systems for immunoglobulin M responses to varicella-zoster virus in reactivated infections (zoster)
Authors: Schmidt N J , Arvin A M
Published: January 2020

An immunoglobulin M response to varicella-zoster virus was detected in 70% of zoster patients by solid-phase radioimmunoassay, in 52% by indirect immunofluorescence, in 48% by neutralization on sucrose density gradient fractions, and in 27% by an antibody class capture enzyme immunoassay. The patients showed marked variations in their varicella-zoster virus immunoglobulin M responses detectable in the different assays.
Rotavirus-associated gastroenteritis in black infants in South Africa
Authors: Steele A D , Alexander J J , Hay I T
Published: January 2020

From March 1983 to February 1985, rectal swabs were collected from the black infants admitted to the gastroenteritis unit at Ga-Rankuwa Hospital, Pretoria, Republic of South Africa, to investigate the prevalence of rotavirus-associated gastroenteritis. Overall, 23.1% of the patients tested were found to be positive for rotaviral illness; a definite seasonal pattern emerged, showing a marked increase in the number positive during the autumn. Other factors of importance in terms of rotaviral illness included the age distribution of the patients, the presence of vomiting, and duration of illness.
Some antigenic properties of Haemophilus parasuis and a proposal for serological classification
Authors: Morozumi T , Nicolet J
Published: January 2020

We propose a serological classification of Haemophilus parasuis into at least five serovars, using an agar-gel-precipitation test with extracts from autoclaved cells. Thirty-two strains were examined, and it was possible to classify 26 of them. The specific antigens were thermostable and soluble and were not affected by pronase treatment but could be extracted by phenol, suggesting a polysaccharide. This polysaccharide seemed to be identical with the capsular substance of serovars 1, 2, and 3. In the presumably uncapsulated strains of serovars 4 and 5, the specific antigen was probably located in the outer membrane. The diversity of the specific substance within the different serotypes was shown by the differences in their electrophoretic migration patterns. Other extraction procedures showed that the washing supernatant and extracts at 60 and 100 degrees C were identical with the 121 degrees C extracts for serovars 1, 2, and 3. In serovars 4 and 5, washing antigens, if present, were different from 100 and 121 degrees C extracts. Other common antigens, presumably proteinaceous antigens, were detected after extraction at 60 and 100 degrees C. The slide and tube agglutination tests allowed classification only for the capsulated strains of serovars 1, 2, and 3. The specific agglutinogens were very sensitive to incubation temperature, and the absorption test showed them to be identical with the 121 degrees C precipitinogens.
Phenotypic characteristics of Aeromonas species isolated from adult humans
Authors: George W L , Jones M J , Nakata M M
Published: January 2020

The phenotypic characteristics of 89 Aeromonas strains, most of which had been isolated from feces, were examined. Eighty-two percent of the isolates could be placed into one of four groups on the basis of five tests. The relationship of these groups to the three motile species of Aeromonas (Aeromonas caviae, A. hydrophila, and A. sobria) that have been isolated from humans is unclear. Because the means for identification of Aeromonas strains to the species level appear to be imprecise and because the role of each of these species in human diarrhea is unclear at this time, we recommend that identification of enteric Aeromonas isolates to the species level not be done routinely.
Colonization of newly arrived house staff by virulent staphylococcal phage types endemic to a hospital environment
Authors: Ballou W R , Cross A S , Williams D Y , Keiser J , Zierdt C H
Published: January 2020

The acquisition of hospital strains of Staphylococcus aureus by new house officers was studied in an 800-bed referral hospital over a 1-year period. S. aureus isolates, including three strains with characteristic phage patterns that had previously been documented to cause disease in patients and colonize hospital personnel, were recovered from the anterior nares of 35 of 54 newly arrived house officers. There was a significant correlation (r = 0.7475; P less than 0.02) between colonization with the dominant hospital strain (S) and exposure to the hospital environment over 12 months. No hospital-wide increase in infections owing to the S strain was seen during this period, which suggests that house staff acquired this strain from reservoirs within the hospital. The finding of colonization with virulent endemic S. aureus strains in house officers working on every ward of the hospital suggests that new strategies for control of S. aureus nosocomial infections must be considered and evaluated.
Enzyme-linked immunosorbent assays for virulence properties of Campylobacter jejuni clinical isolates
Authors: Klipstein F A , Engert R F , Short H B
Published: January 2020

To evaluate the capacity of enzyme-linked immunosorbent assays (ELISAs) to identify pathogenic strains among clinical fecal isolates of Campylobacter jejuni, 40 consecutively obtained strains from 39 sick patients and 1 asymptomatic person were tested by respective ELISAs for enterotoxin production in culture filtrates and for the invasive virulence antigen of bacterial cells. Of the 40 strains, 14 produced the enterotoxin; 15 strains, two of which were also enterotoxigenic, were invasive; and 11 strains had no detectable virulence property. The presence or absence of these virulence properties was confirmed by the demonstration that viable cells of all 12 randomly selected enterotoxigenic or invasive strains tested, but none of 9 nonpathogenic strains tested, caused fluid secretion in rat ligated ileal loops. All 12 patients examined who were infected with an invasive strain had grossly or microscopically evident blood cells in their stools or both, whereas none of those infected with an enterotoxigenic strain had overtly bloody diarrhea, and only 1 of 8 patients examined had microscopically evident blood cells in the stool. Twelve of the invasive, five of the enterotoxigenic, and three of the nonpathogenic strains also produced small amounts of cytotoxin, but there was no correlation between cytotoxin production and an abnormal response in rat ligated ileal loops. These observations show that enterotoxin production or invasiveness or both can be detected by ELISAs in three-fourths of C. jejuni fecal isolates and that there is usually a relationship between the specific pathogenic property of the infecting strain and the clinical mainfestations.
Rapid lysostaphin test to differentiate Staphylococcus and Micrococcus species
Authors: Geary C , Stevens M
Published: January 2020

A rapid, simple lysostaphin lysis susceptibility test to differentiate the genera Staphylococcus and Micrococcus was evaluated. Of 181 strains from culture collections, 95 of 95 Staphylococcus strains were lysed, and 79 of 79 Micrococcus strains were not lysed. The seven Planococcus strains were resistant. Clinical isolates (890) were tested with lysostaphin and for the ability to produce acid from glycerol in the presence of erythromycin. Overall agreement between the methods was 99.2%. All clinical Micrococcus strains (43) were resistant to lysostaphin, and all clinical Staphylococcus strains (847) were susceptible. Seven of the Staphylococcus strains did not produce acid from glycerol in the presence of erythromycin. This lysostaphin test provides results in 2 h. It is easier to perform than previously described lysostaphin lysis methods. It is also more rapid and accurate than the glycerol-erythromycin test.
Growth of Aeromonas species on enteric agars
Authors: Desmond E , Janda J M
Published: January 2020

The efficacy of eight routine enteric agars for supporting the growth of 32 strains of Aeromonas spp. (17 A. hydrophila strains, 8 A. sobria strains, and 7 A. caviae strains) was investigated. The plating efficiency of Aeromonas spp. on these media varied greatly (range, 0 to 100%), as did their colony size when compared with that on noninhibitory medium (5% sheep blood agar). Plating efficiency on seven of these eight media appeared to be strain- and not species dependent. Overall, eosin-methylene blue and Hektoen enteric agars showed low plating efficiencies for A. hydrophila, whereas both A. sobria and A. caviae were severely inhibited on brilliant green agar. When all these species are considered collectively, deoxycholate, MacConkey, and xylose lysine deoxycholate appeared to be the most satisfactory routine agars for Aeromonas spp. recovery when used in conjunction with blood agar.
Specific circulating immune complexes in amoebic liver abscess
Authors: Vinayak V K , Purnima  , Singh K , Venkatwswarlu K , Nain C K , Mehta S K
Published: January 2020

A micro enzyme-linked immunosorbent assay was used to detect the presence of amoebic antigen in polyethylene-glycol-precipitated circulating immune complexes (CIC). A cutoff dilution of greater than 1:160 for the precipitates was taken to be of clinical significance. Among the patients with amoebic liver abscess, 93% (14 of 15 confirmed cases) had indications of amoebic antigen in CIC. In 57% of the suspected cases of amoebic liver abscess, amoebic antigen could be detected in CIC. The specificity of the technique for the demonstration of specific amoebic antigen is indicated by the undetectable levels of amoebic antigen in CIC in serum samples from nine cases of nonamoebic hepatic diseases and 10 apparently healthy subjects. It is thus believed that the specific diagnosis of an individual case of amoebic liver abscess can be made by demonstrating specific amoebic antigen in CIC.
Paleoepidemiologic investigation of Legionnaires disease at Wadsworth Veterans Administration Hospital by using three typing methods for comparison of legionellae from clinical and environmental sources
Authors: Edelstein P H , Nakahama C , Tobin J O , Calarco K , Beer K B , Joly J R , Selander R K
Published: January 2020

Multilocus enzyme electrophoresis, monoclonal antibody typing for Legionella pneumophila serogroup 1, and plasmid analysis were used to type 89 L. pneumophila strains isolated from nosocomial cases of Legionnaires disease at the Veterans Administration Wadsworth Medical Center (VAWMC) and from the hospital environment. Twelve L. pneumophila clinical isolates, obtained from patients at non-VAWMC hospitals, were also typed by the same methods to determine typing specificity. Seventy-nine percent of 33 VAWMC L. pneumophila serogroup 1 clinical isolates and 70% of 23 environmental isolates were found in only one of the five monoclonal subgroups. Similar clustering was found for the other two typing methods, with excellent correlation between all methods. Enzyme electrophoretic typing divided the isolates into the greatest number of distinct groups, resulting in the identification of 10 different L. pneumophila types and 5 types not belonging to L. pneumophila, which probably constitute an undescribed Legionella species; 7 clinical and 34 environmental VAWMC isolates and 2 non-VAWMC clinical isolates were found to be members of the new species. Twelve different plasmid patterns were found; 95% of VAWMC clinical isolates contained plasmids. Major VAWMC epidemic-bacterial types were common in the hospital potable-water distribution system and cooling towers. Strains of L. pneumophila which persisted after disinfection of contaminated environmental sites were of a different type from the prechlorination strains. All three typing methods were useful in the epidemiologic analysis of the VAWMC outbreak.
Imipenem susceptibility testing with a commercially prepared dry-format microdilution tray
Authors: Staneck J L
Published: January 2020

MICs of imipenem, concurrently generated in commercially prepared microdilution trays containing predried antibiotic dilutions (Sensititre), and in a standard agar dilution assay (as recommended by the National Committee for Clinical Laboratory Standards, Villanova, Pa.), were within +/- 1 twofold dilution for 94% of 226 bacterial isolates. Imipenem biological activity remained stable over 5 months of tray storage at room temperature against Pseudomonas aeruginosa ATCC 27853. Activity of imipenem was shown by microdilution testing with 890 clinical isolates to be high, with only 4% of isolates having MICs of greater than or equal to 16 micrograms/ml (in vitro resistance).
11th serogroup of Legionella pneumophila isolated from a patient with fatal pneumonia
Authors: Thacker W L , Benson R F , Wilkinson H W , Ampel N M , Wing E J , Steigerwalt A G , Brenner D J
Published: January 2020

A Legionella-like organism (strain 797-PA-H; ATCC 43130) was isolated from a specimen taken from an endotracheal tube of a patient 4 days before death and from the left lung at autopsy. Growth characteristics were consistent with those for Legionella species. Strain 797-PA-H gave negative test results with available direct immunofluorescence assay conjugates and with slide agglutination test antisera prepared against the 22 Legionella species and 35 serogroups now recognized. Minimal reactivity (1 to 2 +) was observed with both tests by using reagents prepared against the Legionella-like organism Lansing 3. Reciprocal absorption studies, however, showed that the cross-reactive antibodies could be removed easily. Physiologic, gas-liquid chromatographic, and DNA hybridization tests revealed that the strain belonged to the species Legionella pneumophila. Therefore, strain 797-PA-H was designated as the type strain of a new L. pneumophila serogroup, serogroup 11.
Reclassification of Cladosporium bantianum in the genus Xylohypha
Authors: McGinnis M R , Borelli D , Padhye A A , Ajello L
Published: January 2020

It is proposed that the dematiaceous hyphomycete Cladosporium bantianum (Saccardo) Borelli be transferred to the genus Xylohypha (Fries) Mason as Xylohypha bantiana (Saccardo) McGinnis, Padhye, Borelli, et Ajello. This new combination is necessary because X. bantiana produces conidiophores that are indistinguishable from its vegetative hyphae and one-celled, smooth-walled conidia that are borne in long, infrequently branched chains. The blastoconidia do not possess darkly pigmented hila. In contrast, members of the genus Cladosporium Link produce erect, distinct conidiophores and one- to four-celled smooth-to-rough-walled conidia that occur in short, frequently branched, fragile chains. The blastoconidia have darkly pigmented hila. Cladosporium trichoides Emmons is a later synonym of X. bantiana.
Surveillance of mice for antibodies to murine cytomegalovirus
Authors: Anderson C A , Murphy J C , Fox J G
Published: January 2020

The sera of 256 mice from nine commercial sources were screened for antibodies to murine cytomegalovirus (MCMV) because a surveillance of this virus has not been reported in the literature for over a decade. Although no evidence of antibodies to MCMV were detected by complement fixation or nuclear anticomplement immunofluorescence, 54.7% of these sera did have antibodies that were detected by enzyme-linked immunosorbent assay. These data emphasize the need for proper containment of laboratory mice to prevent the potential outbreak of acute MCMV infection. Including MCMV antibody surveillance by enzyme-linked immunosorbent assay in routine health monitoring of mice and imparting these findings in an analysis of the role of MCMV on interpretation of experimental results is advised.
Factors affecting Staphylococcus epidermidis growth in peritoneal dialysis solutions
Authors: McDonald W A , Watts J , Bowmer M I
Published: January 2020

Staphylococcus epidermidis is the most frequent cause of peritonitis complicating continuous ambulatory peritoneal dialysis. We studied factors that might influence the growth of S. epidermidis in commercially available peritoneal dialysis solution (PDS). Test strains were inoculated into PDS and incubated overnight at 37 degrees C. Samples were removed at appropriate intervals, bacterial counts were performed, and growth curves were constructed. We studied the effects of various osmolarities, the neutralization and acidification of fresh and spent PDS, and the effect of intraperitoneal dwell time on the ability PDS to support growth of S. epidermidis. In fresh PDS, numbers of bacteria remained constant after 24 h. No significant differences in growth were observed among PDS with 0.5, 1.5, 2.5, and 4.25% glucose. Neutralizing acidic fresh PDS had no effect on bacterial growth. However, growth did occur in spent PDS. PDS which was recovered after only 2 h in the peritoneal cavity supported growth to the same extent as did PDS recovered after 4 to 6 h. Mean log10 changes after 24 h of incubation were as follows: for fresh PDS, -1.3; after 2 h dwell time, 2.9; after 4 h dwell time, 1.9; and after 6 h dwell time, 1.3. Acidification of spent PDS to less than pH 6.35 produced less rapid growth; mean log10 increases after 24 h of incubation were 1.9 for pH 7.75, 1.6 for pH 6.35, 0.6 for pH 5.75, and 0.7 for pH 4.95. Fresh PDS of all available osmolarities neither supported the growth of S. epidermidis nor was bactericidal. Spent PDS supported bacterial growth, and this growth was partly independent of the neutralization which occurred during the dialysis.
Immunoglobulin class-specific serological responses to adenovirus in respiratory infections of young adult men
Authors: Julkunen I , Lehtomäki K , Hovi T
Published: January 2020

Complement fixation and enzyme immunoassay (EIA) were used for measuring viral antibodies in 103 military conscripts with pneumonia and 98 conscripts with other respiratory infections. Diagnostic rises in antibody to adenovirus were found in 23 (22%) patients with pneumonia and in 42 (43%) patients with other respiratory infections. EIA detected more diagnostic rises than did complement fixation (22 versus 17 for pneumonia and 42 versus 40 in other infections). Adenovirus antibodies were analyzed for different immunoglobulin classes by EIA. Diagnostic rises in immunoglobulin G (IgG) and IgA isotypes were observed in 89 and 77% of the cases, respectively. IgM antibodies were positive in 39% of the cases. In five cases (8%), the demonstration of adenovirus antibodies of the IgM class was the only serological evidence for adenovirus infection. The present study demonstrates that the immunoglobulin class-specific EIA is a sensitive method for the diagnosis of respiratory adenovirus infections.
Morphologic and physiologic studies of three dematiaceous pathogens
Authors: Dixon D M , Salkin I F
Published: January 2020

Uncertainty in identifying a dematiaceous mold from a human bronchial washing precipitated a study of Dactylaria gallopava (Cooke) Bhatt et Kendrick and its relationship to Scolecobasidium constrictum Abbott. Morphologic and physiologic studies were conducted with representative isolates and subcultures derived from the isolates used to prepare the type specimens of these two fungi, as well as a third dark mold, Ochroconis constricta (Abbott) de Hoog et von Arx. All test isolates were morphologically similar in that two-celled, cylindrical blastoconidia with a rhexolytic mode of dehiscence were the predominant and most consistent anamorphic structures. Sympodial development of the conidiophore was too variable to distinguish the isolates. Based on their morphologic similarity, we propose a new combination within the genus Dactylaria as amended by Bhatt and Kendrick, D. constricta (Abbott) Dixon et Salkin. However, D. constricta could be differentiated into two groups on the basis of physiologic tests--a subgroup in which isolates grow at 37 and 45 degrees C, give a delayed positive gelatin reaction, and are inhibited on Mycosel agar; and a second subgroup in which isolates grow on Mycosel agar and give a rapid positive gelatin reaction, but do not grow at 37 or 45 degrees C. We recommend that D. constricta be viewed as a potential pathogen when isolated from clinical specimens.
Identification of Mycobacterium tuberculosis antigens in Seibert fractions by immunoblotting
Authors: Coates S R , Hansen D , Schecter G , Slutkin G , Hopewell P , Affronti L , Echenberg D F
Published: January 2020

Seibert fractions prepared from Mycobacterium tuberculosis culture filtrates were evaluated by immunoblotting with a serum pool from patients with active pulmonary tuberculosis. Antibody activity was observed primarily with antigens in the polysaccharide II and A protein fractions; these fractions were further evaluated by immunoblotting with sera from individual patients with tuberculosis, from individuals without tuberculosis and positive for the purified protein derivative antigen skin test, and from individuals negative for the purified protein derivative antigen skin test. The antigens identified in the protein A fraction, a 32,000-molecular-weight antigen and a heterogeneous high-molecular-weight antigen, reacted with antibody found in sera from all patients with tuberculosis and with antibody from over 25% of the control individuals. A 10,000-molecular-weight antigen, a 30,000- to 44,000-molecular-weight antigen, and a heterogeneous high-molecular-weight antigen were observed in the polysaccharide II fraction; these antigens reacted with serum antibody from 70% or more of the patients with tuberculosis and with antibody from 20 to 70% of the control individuals. One of the antigens, with a molecular weight ranging from 17,000 to 28,000 in the polysaccharide II fraction, reacted with antibody in 64% of the sera from patients with tuberculosis but with only 1 of 15 control normal sera. This antigen may elicit an antibody response specifically associated with tuberculosis.
Characterization of clinical strains of Staphylococcus aureus associated with pneumonia
Authors: Sanford B A , Thomas V L , Ramsay M A , Jones T O
Published: January 2020

A total of 5 Staphylococcus aureus strains from patients with postinfluenzal staphylococcal pneumonia, 7 from burn patients with staphylococcal pneumonia, and 21 from the nasopharynx of carriers were phenotypically characterized. All or most strains produced coagulase, clumping factor, DNase, thermostable DNase, protease, gelatinase, lipase, and pigment; the strains were low to moderate producers of extracellular protein A, fibrinolysin, and alpha-hemolysin. All strains were sensitive to mercury, half were sensitive to arsenate and cadmium, and 67 to 92% were resistant to penicillin. Differences between strains were not statistically significant. Cell surface hydrophobicity was determined by measuring percent adsorption to hexadecane. Hydrophobicity of postinfluenzal staphylococcal pneumonia strains was significantly lower than that of pneumonia strains from burn patients and carriers (P less than 0.005). Immunoblot experiments with sera immune to one clinical test strain allowed the separation of all strains into three groups based on probe-positive reactions with primarily four staphylococcal polypeptides (154,200, 130,000, 77,100, and 64,400 molecular weight). The difference in distribution of clinical and carrier strains was highly significant (P = 0.007).
Preliminary evaluation of a rapid colorimetric method for identification of pathogenic Neisseria
Authors: Hosmer M E , Cohenford M A , Ellner P D
Published: January 2020

A rapid colorimetric method for the identification of pathogenic Neisseria (Identicult-Neisseria; Scott Laboratories, Inc.) based on beta-galactosidase, gamma-glutamylaminopeptidase, and gamma-prolylaminopeptidase is described. All 82 clinical isolates of Neisseria gonorrhoeae, 9 clinical isolates of N. meningitidis, and 5 clinical isolates of N. lactamica were correctly determined to the species level, as were 4 isolates of Branhamella catarrhalis. Reactions were prompt and easily interpreted. The system should be extremely useful in clinical laboratories.
Increased detection of prolylaminopeptidase in Neisseria meningitidis by Identicult-Neisseria
Authors: Sperry J F , Cohenford M A , Campognone P , Lawton W , Chee D O
Published: January 2020

Identicult-Neisseria (Scott Laboratories, Inc., Fiskeville, R.I.), a rapid enzymatic method with chromogenic substrates, was tested in our laboratories for the identification of Neisseria gonorrhoea, Neisseria meningitidis, and Neisseria lactamica. The test correlated very highly in its identification of pathogenic Neisseria spp. with modified New York City fermentation medium. Identicult-Neisseria appeared to be more sensitive in its detection of prolylaminopeptidase activity in N. meningitidis than most of the currently available systems.
Isolation of "Campylobacter hyointestinalis" from a human
Authors: Fennell C L , Rompalo A M , Totten P A , Bruch K L , Flores B M , Stamm W E
Published: January 2020

We isolated "Campylobacter hyointestinalis" from the rectal culture of a homosexual man with proctitis. Phenotypic characterization of the isolate was confirmed by DNA hybridization by using the taxonomic spot blot. "C. hyointestinalis" was previously isolated only from animals but should be considered in the identification of Campylobacter species isolated from humans.
Evaluation and optimization of the DNA filter assay for direct detection of enterotoxigenic Escherichia coli in the presence of stool coliforms
Authors: Yam W C , Lung M L , Ng M H
Published: January 2020

Direct detection of enterotoxigenic Escherichia coli (ETEC) in mixed cultures by DNA filter assay was affected by the presence of other bacteria in the cultures. By shortening incubation to 6 h, bacterial overgrowth was minimized, allowing direct detection of ETEC in stools or mixed cultures initially containing 0.2 to 3% ETEC.
The vector homology problem in diagnostic nucleic acid hybridization of clinical specimens
Authors: Ambinder R F , Charache P , Staal S , Wright P , Forman M , Hayward S D , Hayward G S
Published: January 2020

Nucleic acid hybridization techniques using cloned probes are finding application in assays of clinical specimens in research and diagnostic laboratories. The probes that we and others have used are recombinant plasmids composed of viral inserts and bacterial plasmid vectors such as pBR322. We suspected that there was material homologous to pBR322 present in many clinical samples. because hybridization occurred in samples which lacked evidence of virus by other techniques. If the presence of this vector-homologous material was unrecognized, hybridization in the test sample might erroneously be interpreted as indicating the presence of viral sequences. In this paper we demonstrate specific hybridization of labeled pBR322 DNA with DNA from various clinical samples. Evidence is presented that nonspecific probe trapping could not account for this phenomenon. In mixing experiments, it is shown that contamination of clinical samples with bacteria would explain such a result. Approaches tested to circumvent this problem included the use of isolated insert probes, alternate cloning vectors, and cold competitor pBR322 DNA in prehybridization and hybridization mixes. None proved entirely satisfactory. We therefore emphasize that it is essential that all hybridization detection systems use a control probe of the vector alone in order to demonstrate the absence of material with vector homology in the specimen tested.
Detection of mycobacterial antigens in leprosy serum immune complex
Authors: Patil S A , Sinha S , Sengupta U
Published: January 2020

The antigens from immune complexes of sera from patients with mycobacterial diseases were released by sodium dodecyl sulfate. The antigenic activity of the released proteins was tested by agar gel diffusion and immunoelectrophoresis. This simple method provided direct evidence for the presence of mycobacterial antigens in the immune complexes of sera from patients with leprosy and tuberculosis.
Genetic heterogeneity of recent isolates of adenovirus types 3, 4, and 7
Authors: Bailey A S , Richmond S J
Published: January 2020

Restriction endonuclease analysis was carried out on adenovirus types 3, 4, and 7 (Ad3, Ad4, and Ad7, respectively) isolated during a 16-month epidemic period. Most of the isolates were associated with respiratory or ocular infection or both. Forty-four strains of the subtype Ad7b were identified with SmaI; these strains could be further subdivided into three distinct genome groups, designated Ad7b1, Ad7b2, and Ad7b3, by digestion with PvuII, BglI, and SstII. These Ad7b variants occurred at approximately similar frequencies throughout the epidemic and were associated with similar clinical illnesses. Nine distinct intratypic genome groups were identified among 29 Ad3 strains with HindIII, PstI, and SmaI; adenoviruses similar to subtype Ad3a, formerly reported to be uncommon in Europe, were isolated most frequently (41% of strains), and the previously predominant prototype (Ad3p) was not found. Considerable similarities among the genomes of some strains of Ad3 and Ad7 could be demonstrated, and clinical and epidemiological presentations were very similar. The four strains of Ad4 isolated fell into two distinct patterns, i.e., the previously recognized prototype (Ad4p) and a new subtype designated Ad4b. Both of these subtypes were associated with follicular conjunctivitis. Thus we were unable to produce evidence that particular genome types of Ad3, Ad4, and Ad7 are associated with particular clinical presentations. However, restriction endonuclease analysis should prove useful for epidemiological studies on these viruses.
Rapid dot enzyme immunoassay for the detection of antibodies to cytomegalovirus
Authors: Lin T M , Halbert S P
Published: January 2020

Cytomegalovirus (CMV) antigen was coated onto a white opaque plastic card as small dots inside circles marked in the microtiter plate well pattern. The card with antigen dots could be cut according to the number of test samples to be assayed. Small drops of undiluted serum samples, goat antibodies to human immunoglobulin G labeled with alkaline phosphatase, and finally substrate (5-bromo-4-chloro-3-indolyl phosphate) were sequentially added to the antigen spots and incubated in the open air at room temperature for 5 min each. The antigen dots showed blue color for sera with immunoglobulin G antibodies to cytomegalovirus but no color for those without. The developed antigen dots could be rinsed with water and kept as permanent records. For the assay of a large number of serum samples, a modified procedure with serum diluted 1:10 and longer first two incubations (20 min each) was found to be more comfortable to perform. The results of this assay for 123 undiluted and 256 diluted serum samples revealed very good correlations with those obtained by a commercially available test kit for immunoglobulin G antibodies to cytomegalovirus with 97 and 99% agreement, respectively. This dot test was very reproducible and required no instrumentation. The reagents, including coated antigen dots, are stable at room temperature for at least 2 months and are ready for use.
Purification and characterization of an Aeromonas hydrophila hemolysin
Authors: Asao T , Kozaki S , Kato K , Kinoshita Y , Otsu K , Uemura T , Sakaguchi G
Published: January 2020

A hemolysin produced by Aeromonas hydrophila CA-11, isolated from an environmental source, was purified by sulfopropyl-Sephadex C-25 chromatography at pH 5.0. This hemolysin caused fluid accumulation in infant mouse intestines and rabbit intestinal loops and killed Vero cells, as did the hemolysin produced by strain AH-1, isolated from a diarrheal case. In polyacrylamide gel electrophoreses at pHs 4.0 and 9.4 and in thin-layer isoelectric focusing, CA-11 hemolysin migrated as a single band to a position different from that of AH-1 hemolysin. Immunodiffusion tests indicated that CA-11 hemolysin was immunologically related to AH-1 hemolysin but possessed unique antigenic determinants. Neutralization tests with antihemolysin sera also demonstrated immunological cross-reactivity between AH-1 and CA-11 hemolysins. These results apparently indicate that the hemolysins produced by the two strains of A. hydrophila are immunologically and physicochemically different from each other.
Phenotypic comparison of Pseudomonas aeruginosa strains isolated from a variety of clinical sites
Authors: Woods D E , Schaffer M S , Rabin H R , Campbell G D , Sokol P A
Published: January 2020

Pseudomonas aeruginosa elaborates a number of extracellular products which have been shown to play a role in the pathogenesis of disease caused by this organism. In this study, we showed that the host environment markedly affects the levels of exoproducts produced. We compared the phenotypes of a number of P. aeruginosa strains obtained from a variety of clinical sources, including burn wounds, skin wounds, urine, cystic fibrosis sputum, acute pneumonia sputum, and blood. The clinical isolates were examined quantitatively for levels of total protease, elastase, phospholipase C, exotoxin A, and exoenzyme S produced in vitro under defined conditions. The exoproduct levels varied significantly, depending on the site of isolation. Elevated levels of elastase were demonstrated in strains isolated from acute lung infections, phospholipase C levels were elevated in urinary tract and blood isolates, exotoxin A levels were elevated in blood isolates, and exoenzyme S levels were increased in acute pneumonia isolates. Isolates from cystic fibrosis sputum produced low amounts of virtually all of the tested exoproducts, particularly as compared with sputum isolates from acute P. aeruginosa lung infections.
Alternative cell line for virus isolation
Authors: Smith C D , Craft D W , Shiromoto R S , Yan P O
Published: January 2020

A human lung carcinoma cell line (A549) was compared with various other cell lines to determine susceptibility to viral growth. In the first phase of the study, A549 cells were compared with human embryonic kidney (HEK) and cynomolgus monkey kidney (CMK) cells for isolation of upper-respiratory disease viruses by using 1,248 throat swab specimens from basic-combat trainees. Of the 552 virus isolates, 507 were adenoviruses, 41 were polioviruses, and 4 were herpes simplex viruses (HSV). Of the isolates, 518 (93.8%) were isolated in A549 cells, 480 (87.0%) were isolated in HEK cells, and 262 (47.5%) were isolated in CMK cells (P less than 0.001). In the second phase of the study, A549 cells were compared with a human diploid fibroblast cell strain (MRC-5) and Vero monkey kidney (VMK) cells for the isolation of HSV from 1,157 specimens submitted for culture. Of the 227 HSV isolates, 210 (92.5%) were isolated in A549 cells, 202 (89.0%) were isolated in VMK cells (P greater than 0.1 for A549 versus VMK cells), and 167 (73.6%) were isolated in MRC-5 cells (P less than 0.001 for A549 versus MRC-5 cells). These results suggest that A549 cells are more susceptible to adenovirus infection and at least as susceptible to HSV infection compared with the other cell cultures evaluated. Detracting factors for the use of A549 cells were a slight loss of sensitivity to adenovirus at passage 120 and a concurrent change in the morphology of the cells. The A549 cell line proved to be an efficient, practical, and economical alternative cell system for the isolation of adenovirus and HSV in particular. Initial indications are that other clinically significant viruses may be grown in A549 cells; however, additional studies need to be performed.
Competitive enzyme-linked immunosorbent assay for cholera-related enterotoxins in Salmonella typhimurium
Authors: Hariharan H , Booth B A , Brickman T J , Katt W C , Boesman-Finkelstein M , Finkelstein R A
Published: January 2020

We report a rapid competitive enzyme-linked immunosorbent assay to screen Salmonella typhimurium strains for cholera-related enterotoxin antigens. Polymyxin B extracts of bacterial cells from syncase-glucose broth cultures of 7 of 15 strains gave positive results. The specificity of the test was confirmed with known heat-labile-enterotoxin-positive and -negative Escherichia coli strains which gave significantly different values.
Serum immunoglobulin G antibody subclass responses to respiratory syncytial virus F and G glycoproteins after primary infection
Authors: Wagner D K , Graham B S , Wright P F , Walsh E E , Kim H W , Reimer C B , Nelson D L , Chanock R M , Murphy B R
Published: January 2020

Because the immunoglobulin G (IgG) response to carbohydrate antigens is typically from the IgG2 subclass and the IgG response to protein antigens is typically from the IgG1 and sometimes the IgG3 subclass, two respiratory syncytial virus glycoproteins, F and G, which differ substantially in the amount of glycosylation, were used as antigens in an enzyme-linked immunosorbent assay to determine IgG subclass responses in 20 infants and young children with naturally acquired respiratory syncytial virus infection. Both glycoproteins elicited primarily IgG1 and IgG3 responses, indicating that the protein moieties of the glycoproteins may be immunodominant in this age group.