bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of bacteremia in patients receiving antimicrobial therapy: an evaluation of the antimicrobial removal device and 16B medium
Authors: Doern G V , Gantz N M
Published: January 2020

A total of 1097 blood specimens obtained from patients receiving antibacterial antimicrobial agents were processed by three blood culture systems: standard aerobic and anaerobic radiometric media, resin-containing radiometric medium (16B; Johnston Laboratories, Cockeysville, Md.), and aerobic and anaerobic radiometric media inoculated with blood processed in an Antimicrobial Removal Device (Marion Laboratories, Kansas City, Mo.). A total of 73 cultures, representing 45 unique septic episodes, yielded 81 clinically significant organisms. Forty-six organisms (28 septic episodes) were recovered in standard radiometric medium. 16B medium yielded 63 organisms (37 septic episodes). Sixty-nine organisms (42 septic episodes) were isolated from radiometric blood cultures inoculated with Antimicrobial Removal Device-processed blood. Contamination rates were not significantly different among the three systems. In comparison with standard radiometric blood cultures, the length of time to detection of positive blood cultures was shorter with both 16B medium and with Antimicrobial Removal Device-processed cultures. Comparison of the latter two systems suggested enhanced recovery of clinically significant organisms in radiometric blood cultures inoculated with blood processed in the Antimicrobial Removal Device. There was no difference in the length of time to detection of positive blood cultures.
Modified oxidation-fermentation medium for detection of acid production from carbohydrates by Neisseria spp. and Branhamella catarrhalis
Authors: Knapp J S , Holmes K K
Published: January 2020

A modified oxidation-fermentation medium was developed as a practical medium for highly sensitive and specific detection of acid production from carbohydrates by Neisseria spp. and Branhamella catarrhalis. A total of 756 strains representing 17 Neisseria spp. and Branhamella catarrhalis were tested in this medium, in which the protein concentration was reduced relative to the carbohydrate concentration, phenol red was substituted for bromthymol blue at a low concentration, and the initial pH was adjusted to 7.2. Sugar utilization patterns were consistent with published results and with other cultural and biochemical characteristics for these species. The reactions obtained using this medium were qualitatively better and more reproducible than those obtained in cystine-Trypticase agar (BBL Microbiology Systems, Cockeysville, Md.) medium.
Pediatric viral gastroenteritis during eight years of study
Authors: Brandt C D , Kim H W , Rodriguez W J , Arrobio J O , Jeffries B C , Stallings E P , Lewis C , Miles A J , Chanock R M , Kapikian A Z , Parrott R H
Published: January 2020

During the period January 1974 through July 1982, fecal samples from 1,537 pediatric inpatients with gastroenteritis were tested for enteric viruses by electron microscopic and rotavirus enzyme-linked immunosorbent assay techniques. Rotaviruses were detected in 34.5% of these patients, enteric adenoviruses were detected in 4.7%, approximately 27-nm viruses were detected in 1.6%, and at least one of these agents was found in 40.1% of the study subjects. Three infections were by an apparently new agent which morphologically is a rotavirus, but which failed to react in the rotavirus enzyme-linked immunosorbent assay. During the first 8 calendar years of study, rotaviruses were detected in 39.0% of 577 patients in the even-numbered years and 30.3% of 702 patients in the odd-numbered years. Adenoviruses were found in all calendar months. Rotaviruses were found in inpatients in November through July, whereas approximately 27-nm viruses were found in October through June. The percentage of patients who had a demonstrated viral infection rose steadily from 7.4% in September to 72.0% in January and then steadily declined to 2.9% in August. Viral infection was especially common in study subjects who were 7 through 24 months of age; 61% of such children had one or more enteric viruses. Rotavirus-infected patients tended to be younger during the months of greatest rotavirus activity than at the beginning and end of the rotavirus season, presumably because of a greater exposure to virus at the height of the rotavirus outbreak.
Epidemiology of gentamicin-resistant, gram-negative bacillary colonization in a spinal cord injury unit
Authors: Shlaes D M , Currie C A , Rotter G , Eanes M , Floyd R
Published: January 2020

A prospective epidemiological survey of a spinal cord injury unit for gentamicin-resistant, gram-negative bacilli was undertaken. The initial survey of the unit suggested a low level of cross-infection involving Pseudomonas aeruginosa and Providencia stuartii. However, a longitudinal study of new admissions revealed that only 13 of 52 nosocomial acquisitions could be considered to be due to cross colonization. Comparison of data on antibiotic use did not suggest selective pressure for resistant endogenous flora. Nosocomial acquisition was directly related to the length of the hospital stay. Antibiotic susceptibility testing of gentamicin-resistant, gram-negative bacilli showed only minor differences between nosocomial isolates and those present during the initial survey. Of the usual antimicrobial agents, amikacin, carbenicillin, and cefoxitin were the most active against all organisms, with the exception of Serratia spp. Of the new beta-lactams, ceftazidime and imipemide (N-formimidoyl thienamycin) were most active.
Characterization of clinically significant strains of coagulase-negative staphylococci
Authors: Christensen G D , Parisi J T , Bisno A L , Simpson W A , Beachey E H
Published: January 2020

On occasion, a patient may have two or more clinical cultures yielding a coagulase-negative staphylococcus If these multiple isolates have the same phenotype, one might conclude that the same strain was reisolated from the patient, indicating its persistent and pathological presence. We examined the validity of this conclusion when we applied a number of characterizing systems to a collection of 143 isolates of coagulase-negative staphylococci collected during an outbreak of intravascular catheter-associated sepsis. The probability of classifying two random isolates as the same phenotype or species was as follows: P = 0.356 for phage typing, P = 0.348 for Baird-Parker biotyping, P = 0.346 for the API STAPH-IDENT (Analytab Products) system, P = 0.327 for Bentley et al. biotyping, and P = 0.077 for antimicrobial susceptibility patterns. Although antimicrobial susceptibility patterns had the lowest probability, a variability in test results of 7.7% and a tendency for strains to have similar antibiograms effectively raised the probability to P = 0.897. The combination of the API STAPH-IDENT with antibiograms resulted in a probability of P = 0.037 to P = 0.147. When all of the above methods were used together a probability of P = 0.014 was achieved. Five patients had isolates from two or more blood cultures spaced more than 1 day apart that were identical by all of the above criteria, thus confirming prolonged bacteremia. The collection was also examined for the incidence of slime production. Slime production was not associated with any of the above groups, but was associated with symptomatic infections (P less than 0.05) and gentamicin resistance (P less than 0.01). Slime production was strain stable and was of assistance in typing strains of coagulase-negative staphylococci.
Detection of antibodies to Pseudomonas aeruginosa alginate extracellular polysaccharide in animals and cystic fibrosis patients by enzyme-linked immunosorbent assay
Authors: Bryan L E , Kureishi A , Rabin H R
Published: January 2020

An enzyme-linked immunosorbent assay was developed to detect antibodies to sodium alginate exopolysaccharide purified from three strains of Pseudomonas aeruginosa and commercial alginate from seaweed. Good attachment of alginate occurred with polystyrene microtiter plates at pH 7.0 with 0.04 M sodium phosphate buffer. With the enzyme-linked immunosorbent assay procedure, antibodies to alginate (not previously shown to be immunogenic) could be shown in humans and after immunization of mice and rabbits. Antibody to one of the alginates cross-reacted with two other P. aeruginosa alginates and commercial seaweed alginate. In animal immunization antibody titers were maximal after a single intravenous injection with an optimal dose of P. aeruginosa 3064 alginate. Healthy controls not known to have a previous P. aeruginosa infection had low, but detectable, antibody titers to 3064 and commercial alginate. Three cystic fibrosis patients not colonized with P. aeruginosa had similar antibody levels. Twenty-eight cystic fibrosis patients colonized with P. aeruginosa formed a clearly separate group with antibody titers higher than that of the control and noncolonized cystic fibrosis patients. Antibody titers to 3064 or commercial alginate did not increase during acute P. aeruginosa bronchopneumonitis in 16 cystic fibrosis patients or after repeated episodes in 4 patients.
Direct isolation in cell culture of human rotaviruses and their characterization into four serotypes
Authors: Wyatt R G , James H D , Pittman A L , Hoshino Y , Greenberg H B , Kalica A R , Flores J , Kapikian A Z
Published: January 2020

Of 73 rotavirus-positive fecal specimens tested, 39 yielded a human rotavirus that could be cultivated serially in MA104 or primary African green monkey kidney cells or both; 18 were serotyped. Four distinct serotypes were identified by plaque reduction or tube neutralization assay or both, and three of these serotypes were the same as those established previously by plaque reduction, using human rotaviruses cultivated by genetic reassortment with a cultivable bovine rotavirus. Ten human rotavirus strains received from Japan were found to be similar, if not identical, to our candidate prototype strains representing these four human rotavirus serotypes.
Semiquantitative culture of Gardnerella vaginalis in laboratory determination of nonspecific vaginitis
Authors: Ratnam S , Fitzgerald B L
Published: January 2020

To evaluate the usefulness of quantitative cultures of Gardnerella vaginalis in the laboratory determination of nonspecific vaginitis, the actual and relative numbers of G. vaginalis in genital cultures of a general patient population were assessed semiquantitatively, and the laboratory results were then correlated with the clinical findings. Of the 1,585 women studied, 417 (26.3%) yielded G. vaginalis in culture. Of these, only 113 (27.1%) were found to have symptoms and signs consistent with nonspecific vaginitis. G. vaginalis was obtained in pure or predominant growth from 87 of 100 consecutive cases with nonspecific vaginitis and 32 of 100 consecutive cases without the symptoms or signs of vaginitis (P less than 0.001). Hence, the positive predictive value of isolation of G. vaginalis in pure and predominant growths was determined to be 73% (87 of 119). Conversely, G. vaginalis was isolated in mixed or light growth significantly more often from asymptomatic women than from symptomatic patients, i.e., 68 versus 13 cases. Therefore, the negative predictive value of isolation of G. vaginalis in mixed and light growths was found to be 84% (68 of 81). Quantitation of the relative amount of G. vaginalis growth had higher predictive values as compared with the assessment of G. vaginalis growth alone. We conclude that quantitative culture of G. vaginalis is essential to obtain maximum reliability of culture results in the laboratory determination of nonspecific vaginitis. Although quantitated cultures of G. vaginalis have high predictive values, laboratory results must be interpreted in conjunction with the clinical findings.
Comparison of methods for recovery of Clostridium difficile from an environmental surface
Authors: Buggy B P , Wilson K H , Fekety R
Published: January 2020

Survival of Clostridium difficile in an aerobic environment is possible because of spore formation. When sodium taurocholate is substituted for the egg yolk of a selective medium, cycloserine-cefoxitin-fructose-agar (CCFA), enhanced recovery of C. difficile spores is shown. This selective medium (TCCFA) does not improve recovery of vegetative forms. In this study, dry and saline-moistened swabs, adhesive paddles, and Rodac plates containing CCFA and TCCFA were compared in their ability to recover C. difficile spores from an inoculated surface. Rodac plates grew 20 to 25 times as many spores on TCCFA as on CCFA. Saline-moistened swabs recovered fewer organisms than Rodac plates. Dry swabs and adhesive paddles rarely recovered spores. Prereduction of agar in an anaerobic chamber was not necessary for optimal spore recovery. Optimal growth of vegetative C. difficile required prereduced media. Agar prereduced for 2 h supported the growth of 12 C. difficile isolates as well as agar prereduced for 18 h. Vegetative cells of C. difficile survived for only 15 min in room air. Use of Rodac plates containing TCCFA is preferred for detection of C. difficile spores in the hospital environment.
Evaluation of serum arabinitol as a diagnostic test for candidiasis
Authors: Wells C L , Sirany M S , Blazevic D J
Published: January 2020

Gas-liquid chromatography was used to quantitate the arabinitol concentration in the sera of patients with candidiasis and in that of control patients. Serum arabinitol was elevated in 59% (n = 34) of patients with Candida sepsis, in 39% (n = 38) with Candida colonization, in 14% (n = 62) with bacterial sepsis, and in 0% (n = 11) of normal persons. The above patients were subsequently divided on the basis of renal function. Of those with decreased renal function, serum arabinitol was elevated in 89, 92, and 50% of patients with Candida sepsis, Candida colonization, and bacterial sepsis, respectively. Of those with normal renal function, serum arabinitol was elevated in only 23 and 14% of patients with Candida sepsis and Candida colonization, respectively. When serum arabinitol/creatine ratios were calculated for patients with both increased arabinitol and increased creatinine, elevated ratios were obtained in 69, 36, and 0% of patients with Candida sepsis, Candida colonization, and bacterial sepsis, respectively.
Evaluation of sodium polyanethanol sulfonate as a blood culture additive for recovery of Salmonella typhi and Salmonella paratyphi A
Authors: Escamilla J , Santiago L T , Uylangco C V , Cross J H
Published: January 2020

A total of 640 blood specimens from patients in an area endemic for enteric fever were cultured in parallel in tryptic soy broth with and without sodium polyanethanol sulfonate (SPS). A total of 95 specimens were positive for Salmonella spp., 54 for Salmonella typhi, and 41 for Salmonella paratyphi A in one or both bottles of a set. Significantly higher rates of recovery were obtained from the SPS-containing medium (P less than 0.01) upon subculturing blindly at 24 h and 3 days of incubation. Subcultures performed at 7, 14, and 21 days also yielded a greater number of positive cultures with SPS than without it, although the differences between the two media were not significant (P greater than 0.05). Neither of the media yielded 100% of the positive cultures. Moreover, even if the results of the two media were combined, 34 and 19% of the isolates would have been missed if the specimens had not been incubated to 14 and 21 days, respectively. The data indicate that SPS aids in early recovery of S. typhi and S. paratyphi A from blood cultures, and additionally, that under the conditions used in the study, incubation beyond a 1-week period is required for efficient isolation of these organisms from blood.
Presumptive identification of Vibrio species with H antiserum
Authors: Tassin M G , Siebeling R J , Roberts N C , Larson A D
Published: January 2020

Species in the genus Vibrio exhibit flagellar (H) antigens unique to the species. Thus, species-specific H antiserum could be a valuable reagent with which to screen serologically large numbers of Vibrio isolates. Antisera against V. cholerae, V. fluvialis, V. anguillarum, V. metschnikovii, V. parahaemolyticus, V. alginolyticus, and V. vulnificus H antigens was produced in rabbits by repeated injections of Formalin-preserved whole cells. Anti-O activity and anti-H activity against common H antigens was absorbed from each antiserum, V. fluvialis was shown to possess an H antigen unique to the species and also to share minor H antigens with V. cholerae, V. metschnikovii, and V. anguillarum. V. vulnificus also exhibits a species-unique H antigen. A comprehensive serological screening system based on species-specific H antiserum was developed to identify pathogenic Vibrio species one step beyond primary isolation. Vibrio species were correctly identified with accuracies ranging from 93 to 100%. Some isolates were either nonmotile or poorly so and thus did not flocculate in H antiserum.
Evaluation of the automicrobic system gram-positive identification card for species identification of coagulase-negative staphylococci
Authors: Almeida R J , Jorgensen J H , Johnson J E
Published: January 2020

The AutoMicrobic system Gram-Positive Identification Card (Vitek Systems, Inc., Hazelwood, Mo.) was evaluated for identification of a group of 150 isolates of coagulase-negative staphylococci. Identifications obtained with the Gram-Positive Identification Card were compared with reference identifications derived from 15 conventional biochemical tests. The AutoMicrobic system correctly identified only 67.3% (101 of 150) of the test isolates. The greatest accuracy was achieved with Staphylococcus epidermidis isolates (95.7%), whereas Staphylococcus hominis isolates were least often correctly identified (26.7%).
Comparative analysis of serum antibody responses to Pseudomonas aeruginosa exotoxin A by cystic fibrosis and intensive care unit patients
Authors: Cukor G , Blacklow N R , Nowak N A , Rich C M , Braverman L E , Fischer R A
Published: January 2020

Pulmonary infection with Pseudomonas aeruginosa is a major cause of morbidity and mortality in cystic fibrosis (CF) patients. P. aeruginosa toxin is one of several proposed virulence factors which may be responsible for chronic P. aeruginosa infections in these patients. With a highly specific, sensitive, and quantitative radioimmunoassay (RIA) and a cell culture assay, the humoral immune responses of CF patients in terms of total antitoxin, antitoxin immunoglobulins A and M, and neutralizing antitoxin were compared with those of P. aeruginosa-infected intensive care unit patients and controls. The P. aeruginosa-infected CF patients were divided into severe and moderate disease groups based on mortality observed over an 8-year period. The intensive care unit patients were divided by the site of infection and the controls were healthy children and uninfected CF patients. Antibodies to toxin were found in the sera of all subjects by radioimmunoassay. Neutralizing antibody was associated with current infection. Elevated titers of antitoxin immunoglobulin A were found only in subjects with pulmonary P. aeruginosa infections. No significant differences in any antibody class were observed between the severe and moderate disease groups. In addition, no differences were observed in the antitoxin immune response of chronically infected CF patients and intensive care unit patients with acute pulmonary infections.
Evaluation of a lysis-centrifugation system for recovery of yeasts and filamentous fungi from blood
Authors: Bille J , Stockman L , Roberts G D , Horstmeier C D , Ilstrup D M
Published: January 2020

A lysis-centrifugation system (Isolator; E. I. du Pont de Nemours & Co., Inc., Wilmington, Del.) was compared with a biphasic brain heart infusion (BHI) medium in a prospective study of 5,125 fungal blood cultures. The Isolator recovered 90.3% of the positive cultures, compared with 63.4% recovered by the biphasic BHI medium. Overall, the detection of fungemia was increased 36.6% by the Isolator. Mean recovery times for yeasts were 2.12 and 4.90 days for the Isolator and BHI bottles, respectively. Cultures of Histoplasma capsulatum required 8.0 and 24.14 days for recovery by the Isolator and BHI bottles, respectively. The Isolator provided a more rapid and sensitive means of detecting organisms associated with fungemia.
Rapid species identification and biotyping of respiratory isolates of Haemophilus spp
Authors: Matthews J S , Reynolds J A , Weesner D E , Perry J L , Jenkins A L
Published: January 2020

Three commercially available systems, the 4-h Minitek Enterobacteriaceae III, the Haemophilus Trio-Tube, and the Micro-ID, were evaluated for their capacities to identify and biotype 308 respiratory isolates of Haemophilus spp. When compared with aminolevulinic acid test results, the definitive identification method used in this study, these systems demonstrated no significant differences in their capacities to differentiate Haemophilus influenzae from Haemophilus parainfluenzae. They were in agreement with the standard method of species identification approximately 50% of the time. When sucrose was added to the Minitek and Trio-Tube configurations, the efficiency rate of species identification increased to more than 95%. The Micro-ID could not be modified to incorporate this additional biochemical parameter. The performance of the sucrose-supplemented Minitek and Trio-Tube systems, compared to the combined results of Micro-ID and aminolevulinic acid, produced correlations of 94 and 90%, respectively. Rapid and accurate methodologies are available for combined species identification and biotyping of Haemophilus spp.
Differentiation between virulent and avirulent Yersinia enterocolitica isolates by using Congo red agar
Authors: Prpic J K , Robins-Browne R M , Davey R B
Published: January 2020

Cultivation of clinical isolates of Yersinia enterocolitica of diverse geographical origin on a medium containing 5 micrograms of Congo red per ml disclosed two colony types. These were designated CR+ and CR- according to their ability to bind Congo red. CR+ strains bore plasmids of between 40 and 50 megadaltons and were positive in several tests of Y. enterocolitica virulence, including autoagglutination, reduced growth on magnesium oxalate agar, resistance to the bactericidal effect of serum, and lethality for iron-overloaded mice. CR- strains were plasmidless and were negative in all these assays. The Congo red reaction provides a simple and efficient means of screening Y. enterocolitica for virulence and is the best available method for identifying individual plasmid-bearing colonies.
Induction of cross-reactive serum neutralizing antibody to human rotavirus in calves after in utero administration of bovine rotavirus
Authors: Wyatt R G , Kapikian A Z , Mebus C A
Published: January 2020

Sera from calves infected in utero or postnatally with bovine rotavirus NCDV or postnatally with human rotavirus D (serotype 1) were tested by plaque reduction neutralization assay for antibody to bovine rotavirus and to three serotypes of human rotavirus. Homologous antibody developed in all animals, but antibody to heterologous rotaviruses developed mainly in animals exposed in utero to bovine rotavirus. The development of heterologous antibody may explain the immunological implications for cross-protection, previously observed between bovine and human rotavirus in experimentally infected calves.
Evaluation of the MS-2 urine screening system
Authors: Smith J A , Ngui-Yen J
Published: January 2020

A total of 1,736 urine specimens were evaluated in a two-phase study, and the results were compared with those obtained by the standard semiquantitative culture plate method. In phase 1, 1,083 urine specimens were examined by utilizing modified broth containing 0.03% agar, whereas 653 specimens in phase 2 were examined after preincubation of the same broth overnight at 35 degrees C. In both phases of the study, more than one-half of the total urine samples (54%) were negative (less than or equal to 10(3) CFU/ml), and approximately 16% had colony counts of less than or equal to 10(5) per ml. The sensitivity and specificity in phase 1 were 99.5 and 70.5%, respectively, compared with 92.6 and 83.8% in phase 2. Preincubating the urine ampvettes at 35 degrees C reduced the false-positive rate from 21.9 to 10.5%. The performance requisites of acceptable sensitivity and specificity appeared to be met by the MS-2 (Abbott Laboratories, Diagnostics Div., Dallas, Tex.) screening method.
Laboratory investigation of hemorrhagic colitis outbreaks associated with a rare Escherichia coli serotype
Authors: Wells J G , Davis B R , Wachsmuth I K , Riley L W , Remis R S , Sokolow R , Morris G K
Published: January 2020

Two outbreaks of hemorrhagic colitis, a newly recognized syndrome characterized by bloody diarrhea, severe abdominal pain, and little or no fever, occurred in 1982. No previously recognized pathogens were recovered from stool specimens from persons in either outbreak. However, a rare E. coli serotype, O157:H7, was isolated from 9 of 20 cases and from no controls. It was also recovered from a meat patty from the implicated lot eaten by persons in one outbreak. No recovery of this organism was made from stools collected 7 or more days after onset of illness; whereas 9 of 12 culture-positive stools had been collected within 4 days of onset of illness. The isolate was not invasive or toxigenic by standard tests, and all strains has a unique biotype. Plasmid profile analysis indicates that all outbreak-associated E. coli O157:H7 isolates are closely related. These results suggest that E. coli O157:H7 was the causative agent of illness in the two outbreaks.
Comparison of immunodiffusion techniques with standard complement fixation assay for quantitation of coccidioidal antibodies
Authors: Wieden M A , Galgiani J N , Pappagianis D
Published: January 2020

Quantitative immunodiffusion (QID) and complement fixation (CF) methods were compared for their agreement in detecting coccidioidal antibodies. For these studies, we assayed 719 sera from 181 patients with coccidioidomycosis. Over 60% of the specimens had CF results of 1:2 to 1:256. A total of 43 patients had five or more specimens obtained over periods of between 1 and 8 years. The QID method, as originally performed, agreed within a twofold dilution of the CF titer in 191 of 267 sera (71.5%). Modification of QID by repeated filling of the antigen and serum wells improved agreement to 84.7% (383 of 452 sera). The degree of CF titer change in patients over time periods was more closely matched by the modified than by the original QID method. Discrepancies between the CF and QID methods appeared not to be due to a subpopulation of patients. QID measurement of coccidioidal antibodies may be a useful substitute for the CF assay in certain clinical laboratories.
Immunoassay for detecting Chlamydia trachomatis major outer membrane protein
Authors: Caldwell H D , Schachter J
Published: January 2020

The development of a solid-phase immunoassay for the detection of the 39,500-dalton major outer membrane protein of the Chlamydia trachomatis lymphogranuloma venereum serotype L2 is described. The test uses immunoadsorbent-purified rabbit anti-L2 major outer membrane protein immunoglobulin G (IgG) passively adsorbed to microtiter plates as a capture antibody. This same IgG antibody was either conjugated to horseradish peroxidase or radioiodinated with 125I and used as a probe to detect major outer membrane protein bound to immobilized IgG. At its greatest sensitivity, the test was capable of detecting 0.5 to 1 ng of purified major outer membrane protein, 5 X 10(3) elementary body inclusion-forming units, and approximately 100 C. trachomatis intracytoplasmic inclusions per assay.
Rapid detection and identification of herpes simplex virus in cell culture by a direct immunoperoxidase staining procedure
Authors: Miller M J , Howell C L
Published: January 2020

Cell monolayers were inoculated with 169 fresh and 76 previously frozen clinical specimens and examined for the presence of herpes simplex virus (HSV) by noting the appearance of characteristic cytopathic effect (CPE) and by using direct immunoperoxidase (IP) stain for viral antigen. HSV was detected by IP staining in 40 of 169 (23.7%) monolayers and by CPE in 39 of 169 (23.1%) monolayers inoculated with fresh specimens. All 40 isolates were detected and confirmed by IP staining within 24 h. Although 39 of 40 isolates were detected by CPE, only 9 of 39 (23%) were positive within 24 h. CPE was observed at 2.7 days on the average, but 4 days were required before 90% of the cultures were positive and more than 5 days were required before all HSV isolates were recognized. Similar results were observed for frozen specimens. HSV was detected earlier with IP staining, which demonstrated more extensive infection of cell monolayers inoculated with titrated fresh culture isolates and clinical specimens than did CPE. IP staining reduces the amount of time required for detection and identification of HSV in culture, is readily adaptable for use in the clinical laboratory, and permanent stained preparations can be made.
Evaluation of a lysis-centrifugation and biphasic bottle blood culture system during routine use
Authors: Kelly M T , Buck G E , Fojtasek M F
Published: January 2020

An in-use evaluation of a commercially available lysis-centrifugation blood culture system (Isolator; Du Pont Co., Wilmington, Del.) is presented. The Isolator was compared with biphasic bottles containing Trypticase soy broth and agar for the detection of organisms in 3,129 paired blood samples. Of 272 potential pathogens recovered, 78% were detected by the Isolator system, and 69% were detected by the biphasic bottle. A total of 31% of these organisms were detected only by the Isolator, and 22% were detected only by the biphasic bottle. The Isolator demonstrated enhanced detection of facultative gram-negative bacilli, anaerobic bacteria, and polymicrobial cultures. The biphasic bottle was more effective for the recovery of facultative gram-positive cocci, especially Streptococcus pneumoniae. The two systems were equally effective for the recovery of yeasts. Contamination rates were 3% for the Isolator and 3.2% for the biphasic bottle. The results indicate that the Isolator system performs well in routine clinical use, but it should be complemented by another method to obtain optimal detection of bacteremia. The biphasic bottle provides an acceptable complementary system both in terms of utility and performance.
Rapid identification of pregnant women heavily colonized with group B streptococci
Authors: Jones D E , Friedl E M , Kanarek K S , Williams J K , Lim D V
Published: January 2020

Pregnant women admitted to Tampa General Hospital, Tampa, Fla., were cultured for group B streptococci (GBS). Culture swabs were placed into enriched, selective Todd-Hewitt medium and were quantitated for GBS. The broth cultures were tested by slide coagglutination before incubation and after 5 and 20 h of incubation. Fifty-four (27%) of the 201 maternity patients cultured were positive for GBS and were identified as such by slide coagglutination. A strong correlation was found between the magnitudes of colonization and the times required to identify the broth cultures as GBS positive. Cultures from mothers heavily colonized (mean concentrations of 3 X 10(4) GBS per culture swab or greater) were identified after 5 h or less of incubation. Mothers lightly colonized with GBS (mean concentrations of 2 X 10(2) GBS per culture swab) were identified only after their broth cultures had been incubated for 20 h.
Urinary adenylate kinase and urinary tract infections
Authors: Bernstein L H , Horenstein J M , Russell P J
Published: January 2020

Positive correlations were found among the appearance of adenylate kinase activity in the urine and the existence of bacteriuria, the Fairley test, and other criteria of urinary infection. Since the adenylate kinase isozymes of human tissues are organ specific and can be distinguished from one another, the appearance of adenylate kinase isozymes in urine was used in this study to identify the existence of infection in bladder or kidney. The findings suggest the usefulness of measuring the appearance of urinary adenylate kinase isozymes for the purpose of detection and differential diagnoses of urinary infections, particularly since adenylate kinase is absent or found in low concentrations in urine and serum under normal conditions.
Effect of different media on determination of novobiocin resistance for differentiation of coagulase-negative staphylococci
Authors: Goldstein J , Schulman R , Kelley E , McKinley G , Fung J
Published: January 2020

Species identification of coagulase-negative staphylococci often requires the determination of novobiocin susceptibility. Although previous investigators have recommended the use of P agar for this purpose, most clinical laboratories do not routinely utilize this medium. For this reason, studies were performed to compare novobiocin susceptibility results obtained with 11 different species of staphylococci (10 isolates of each species), using P agar, Trypticase soy agar with 5% sheep blood, and Mueller-Hinton agar. Tests performed on 70 susceptible isolates (minimal inhibitory concentration less than 1.6 micrograms/ml) resulted in zones of inhibition around 5-micrograms novobiocin disks ranging from 19.6 to 33.9, 16.2 to 26.6, and 21.3 to 36.4 mm on P agar, Trypticase soy agar with 5% sheep blood, and Mueller-Hinton agar, respectively. Forty resistant isolates (minimal inhibitory concentration greater than or equal to 1.6 micrograms/ml) exhibited zones of inhibition ranging from 6.0 to 11.3 mm on P agar, 6.0 to 11.6 mm on Trypticase soy agar with 5% sheep blood, and 6.0 to 13.5 mm on Mueller-Hinton agar. Using the established cut off of 16 mm to define novobiocin resistance for the identification of coagulase-negative staphylococci, we correctly identified 100% of the strains tested, regardless of the media utilized.
Nylon bead enzyme-linked immunosorbent assay for detection of sub-picogram quantities of Brucella antigens
Authors: Perera V Y , Creasy M T , Winter A J
Published: January 2020

An indirect sandwich enzyme-linked immunosorbent assay, using antibody covalently coupled to nylon beads, has been adapted for the detection of Brucella antigens. Optimum conditions were achieved by incubation of 1 ml of reaction mixture with a single bead, and by minimizing nonspecific interactions through the use of beads coated with purified bovine antibodies, preabsorption of third layer rabbit antibodies with normal bovine serum, and treatment of beads with normal goat serum before addition of the goat anti-rabbit enzyme conjugate. Beta-galactosidase was selected for use with clinical samples primarily because of low levels of endogenous enzyme in bovine leukocytes. Use of a fluorogenic substrate enhanced sensitivity 20-fold. Under these conditions, 100 fg of solubilized crude lipopolysaccharide or 8 to 10 Brucella cells was detectable in a fixed volume of 1 ml. A system was also devised for concentrating antigen which permitted ready detection of 2 pg of lipopolysaccharide in a volume of 50 ml (40 fg/ml). Attempts to detect lipopolysaccharide in the presence of concentrated serum or plasma were unsuccessful, but 10 brucellae added to a suspension of leukocytes from 100 ml of normal bovine blood were easily measured.
Immunofluorescent examination of the skin of rabies-infected animals as a means of early detection of rabies virus antigen
Authors: Blenden D C , Bell J F , Tsao A T , Umoh J U
Published: January 2020

Correlations were made on immunofluorescence positivity to antirabies conjugate between cranium-derived nerve fibers in skin and traditional samplings of brain tissue from several species and illness categories of animals with naturally acquired rabies. The overall correlation of results from all categories was about 98% (n, 104) for those that were brain positive and 100% (n, 99) for those that were brain negative. Some animals that ultimately developed rabies were found to have immunofluorescence-positive results 2 or more days before the onset of clinical signs in both natural and experimental infections. The percentage of those with positive skin immunofluorescence results increased as the onset of symptoms approached. From the midcourse period of illness to death, the correlation between skin and brain approached 100%. Different vaccines, commonly given to prevent rabies and other diseases of dogs and cats, were administered to groups of mice and were found to not produce false-positive results when their skin was examined by immunofluorescence for rabies virus antigen. These data suggest that examination of surgical biopsy specimens by immunofluorescence for rabies virus antigen is a useful and reliable diagnostic tool to evaluate the rabies status of biting dogs or cats, or to confirm a clinical diagnosis of rabies in the species tested. The biopsy evaluation of any other species as a means of assessing bite risk is not suggested by these data.
Use of monoclonal antibodies to human immunoglobulin M in "capture" assays for measles and rubella immunoglobulin M
Authors: Forghani B , Myoraku C K , Schmidt N J
Published: January 2020

Monoclonal antibodies to human immunoglobulin M (IgM) were used in a four-phase enzyme immunofluorescence "capture" assay for determination of IgM antibodies to measles and rubella viruses. Little or no background reactivity was seen in the test system, and interfering effects of rheumatoid factor were avoided by preabsorption of test sera with aggregated human IgG. Virus-specific IgM antibody was demonstrable in 23 of 24 patients with serological evidence of measles virus infections and in 36 of 36 patients with serological evidence of postnatal rubella infection. A few of the rubella patients did not show IgM antibody until 5 days after onset of illness. The enzyme immunofluorescence assay was able to demonstrate rubella IgM antibody in congenitally infected newborns, whereas indirect immunofluorescence results for virus-specific IgM were negative. Viral IgM antibody was not detected in persons with past infections with the test viruses, in young children without evidence of past infection, or in patients infected with heterotypic viruses, rickettsiae, chlamydiae, or mycoplasmas.
Rotavirus as a cause of severe gastroenteritis in adults
Authors: Echeverria P , Blacklow N R , Cukor G G , Vibulbandhitkit S , Changchawalit S , Boonthai P
Published: January 2020

Rotavirus was identified as the only etiological agent in 5% of adults (28 of 526) with diarrhea who were admitted to Bamrasnaradura Hospital in Nonthaburi, Thailand, during a 1-year period. Infection was determined by detection of rotavirus in diarrheal stools by enzyme-linked immunosorbent assay accompanied by a greater than fourfold rise in serum complement fixation and radioimmunoassay antibody titers to rotavirus. Adults with clinical rotavirus infections were as severely ill as patients with most bacterial enteric infections; only patients with cholera passed more watery stools and were more dehydrated than those with rotavirus infections. Only 2 of the 28 adults with rotavirus infections had known recent contact with young children with diarrhea. Rotavirus infections in these adults occurred most frequently in the cooler, drier months in Thailand than during the rest of the year. In some settings, rotavirus should be considered in the differential diagnosis of severe diarrhea in adults as well as in young children.
Environmental study of a methicillin-resistant Staphylococcus aureus epidemic in a burn unit
Authors: Rutala W A , Katz E B , Sherertz R J , Sarubbi F A
Published: January 2020

During an outbreak of infections caused by methicillin-resistant (MR) Staphylococcus aureus in our burn unit, we conducted an extensive 10-week study to define the environmental epidemiology of the organism. The inanimate environment in patient rooms and adjacent areas was examined by using volumetric air samplers and Rodac plates. Airborne and surface level contamination with MR S. aureus was quantitated, and overall, MR S. aureus comprised 16, 31, and 40% of all bacterial growth from air, elevated surfaces, and floor surfaces, respectively. Mean air, elevated surface, and floor surface MR S. aureus contamination in rooms of MR S. aureus-infected burn patients were 1.9 MR S. aureus per ft3 (ca. 0.028 m3), 20 MR S. aureus per Rodac plate and 48 MR S. aureus per Rodac plate, respectively. Peak patient room environmental contamination levels were 6.9 MR S. aureus per ft3 of air, 70 MR S. aureus per Rodac plate per elevated surface and 138 MR S. aureus per Rodac plate per floor surface. Environmental contamination levels in the adjacent work areas were considerably lower than in infected patient rooms. There was ample opportunity for contamination of personnel through the inanimate environment in this unit.
Evaluation of a two-minute test for urine screening
Authors: Pezzlo M T , Wetkowski M A , Peterson E M , de la Maza L M
Published: January 2020

A study was conducted to evaluate the ability of a urine filtration system (Bac-T-Screen, Marion Laboratories, Inc., Kansas City, Mo.) to detect negative urine cultures within 2 min. A total of 1,000 urine specimens were tested with the Bac-T-Screen and compared with a standard semiquantitative culture plate method and the Autobac system (General Diagnostics, Warner-Lambert Co., Morris Plains, N.J.). Of the 1,000 clean voided urine specimens tested, 246 specimens had colony counts greater than or equal to 10(5) CFU/ml by the culture plate method. Of these, the Bac-T-Screen detected 65.4% (161 of 246), and the Autobac detected 63.0% (155 of 246). When pure cultures of diphtheroids, lactobacilli, and viridans streptococci other than group D and cultures containing multiple organisms were considered to be contaminants and, therefore, were excluded, there were 106 pure cultures of probable pathogens of which the Bac-T-Screen detected 76.4% (81 of 106) and the Autobac detected 90.6% (96 of 106). Some 133 specimens were uninterpretable with the Bac-T-Screen because 36 clogged the filter and 97 left a residual pigment on the filter. A majority of those clogging the filter (69.4%) had positive plate counts, whereas the majority of the pigmented urines had negative plate counts. Of those urine specimens tested. 754 were negative by the culture plate method. The false-positive rates for Bac-T-Screen and Autobac were 16.2 and 5.8%, respectively. As a urine screen, the Bac-T-Screen has a negative predictive value comparable to the Autobac system and has the advantage of being a 2-min test.
Nocardia asteroides recovery from a dog with steroid- and antibiotic-unresponsive idiopathic polyarthritis
Authors: Buchanan A M , Beaman B L , Pedersen N C , Anderson M , Scott J L
Published: January 2020

This report describes a fatal case of idiopathic polyarthritis in a dog that was partially responsive to vigorous immunosuppressive treatment. Synovial fluids were cultured for L-forms at the following stages of disease: (i) acute arthritic relapse, (ii) incomplete remission, and (iii) death. Nocardia asteroides UCD 1-581 was recovered from the L-form broth culture of the specimen taken during acute relapse, 5 weeks after inoculation, but not at any other stage of disease. Numerous conventional microbiological cultures were unproductive during all phases. Changes occurring in L-form plates included the formation of large irregular mineral deposits and many transferable bodies resembling pseudocolonies. Microscopic examination revealed the presence of many intracellular golden-brown granules and acid-fast bodies in macrophages of the lung and bronchial lymph node tissues. The granules are believed to be the variants embedded in calcium deposits similar to those which developed in the L-form cultures in vitro. Fluorescence of these acid-fast bodies with antibody specific for superoxide dismutase of N. asteroides GUH-2 and labeled anti-immunoglobulin G established their relationship to the isolate. The unrelenting course of disease and the persistence of N. asteroides as an L-form in this animal despite vigorous immunosuppression suggest that this organism plays a direct role in the etiology of this disease.
Primary peritonitis due to Haemophilus influenzae type b in a previously healthy child
Authors: Chang M J , Controni G
Published: January 2020

A case of primary peritonitis caused by Haemophilus influenzae type b biotype 2 in a 3-year-old child is described. The organism was isolated from peritoneal fluid. This is the first case of documented peritoneal infection due to this species in a patient who showed no evidence of being immunocompromised.
Inoculum preparation for anaerobic susceptibility tests
Authors: Murray P R , Niles A C
Published: January 2020

The results of anaerobic susceptibility tests performed with inocula prepared directly from agar isolation media and from overnight broth cultures were compared. Altogether, 93.0% of the results with these two inoculum preparations were within one twofold dilution, including 92.5 and 95.6% of the results with Clostridium species and the Bacteroides fragilis group, respectively. Thus, inocula prepared from agar subculture plates resulted in more-timely susceptibility test results. In addition, the test organisms used in this study grew better when the inoculum was prepared directly from agar plates.
Mycoplasma gallisepticum in culture with biosilon microcarrier beads
Authors: Freidlin P J
Published: January 2020

Various dilutions of Mycoplasma gallisepticum were cultured in the presence of Biosilon microcarrier beads. The microcarriers did not affect the recoverability or the growth rate of M. gallisepticum. Cultures attained a higher density in the presence of microcarriers. The initiation of a culture could be accomplished by the transfer of one bead from a microcarrier culture.
Use of enzyme immunoassays and the latex agglutination test to measure the temporal appearance of immunoglobulin G and M antibodies after natural infection or immunization with rubella virus
Authors: Meegan J M , Evans B K , Horstmann D M
Published: January 2020

The time course of appearance of antibodies after infection with rubella virus was determined with an immunoglobulin G (IgG) detection enzyme-linked immunosorbent assay, a latex agglutination test, and an IgM detection enzyme-linked immunosorbent assay. In six naturally infected rubella patients and 26 vaccinees, antibodies measured by either the IgG enzyme-linked immunosorbent assay or the latex agglutination test generally appeared in parallel with those detected by the hemagglutination inhibition test. By 28 days after inoculation of live virus vaccine and by 2 days postonset of clinical rubella symptoms caused by natural infection, antibodies were found by the two tests for all individuals. A commercially available enzyme-linked immunosorbent assay kit was used to detect rubella-specific IgM. After natural infection, IgM appeared earlier than IgG, and although IgM titers decreased rapidly postinfection, in four of five patients antibodies were still detectable 40 to 43 days after the onset of clinical symptoms. After vaccine-induced infection, rubella-specific IgM was lower in titer than after natural infection and was detected in only three of seven vaccinees 70 days post-immunization.
Reactivity of nonsensitized control erythrocytes in rubella passive hemagglutination assays
Authors: Madsen R D , Coates S R , Rippe D F
Published: January 2020

Two commercially available, nonsensitized control erythrocytes for use in rubella antibody passive hemagglutination assays detected nonspecific reactions in 10 of 600 (1.7%) and 9 of 500 (1.8%) sera tested. Reactive sera were positive with one or the other cell type, but not both. The probability of obtaining a false-positive result owing to a nonspecific passive hemagglutination reaction was estimated to be 0.17% for either cell type.