bluuebunny/crossref_metadata_2025 huggingface dataset:
Rapid identification of material colonization with group B streptococci by use of fluorescent antibody
Authors: Boyer K M , Gadzala C A , Kelly P C , Burd L C , Gotoff S P
Published: January 2020

To identify women colonized with group B streptococci during parturition, we used pooled type-specific fluorescent antibody to examine vaginal swabs enriched by preincubation in selective broth medium. In preliminary experiments, group B streptococcus strain III-Bell was reliably detectable with fluorescent antibody at concentrations of greater than 10(5) colony-forming units per ml, achieved after 6 h of incubation of small inocula (18 to 26 colony-forming units). Of the vaginal swabs from 924 parturient women examined prospectively by both fluorescent antibody and selective bacteriology techniques, group B streptococci were isolated in 154. The sensitivity of the fluorescent antibody technique increased with increasing incubation time and ranged from 49% (3 to 6 h) to 81% (7 to 12 h) to 83% (13 to 18 h) to 93% (greater than 18 h). Colonized mothers identified within 6 h by the fluorescent antibody technique had higher rates of vertical transmission to their newborn infants (61%) than colonized mothers whose fluorescent antibody examinations were negative within this time interval (32%; P = 0.027). However, because of the timing of their admissions, none of the colonized mothers of the four infants who developed early-onset group B streptococcal sepsis were identified with fluorescent antibody until after delivery. Although its sensitivity approaches selective culture methods after 6 h of incubation, fluorescent antibody examination of vaginal swabs does not appear to offer a practical approach to identifying colonized parturient women for intrapartum antibiotic prophylaxis of group B streptococcal infection.
Biochemical characterization of Haemophilus species with the minitek differentiation system
Authors: Kawakami Y , Okimura Y , Kanai M
Published: January 2020

Characterization of 102 Haemophilus strains by the Minitek differentiation system (BBL Microbiology Systems, Cockeysville, Md.) was compared with that by the conventional method. The results obtained by the two methods were in good agreement (P less than 0.001; test of independence). Therefore, this system was found to be applicable for the identification of Haemophilus species in routine clinical bacteriology.
Hemagglutinin Typing as an Aid in Identification of Biochemically Atypical
Escherichia coli
Strains
Authors: Crichton Pamela B. , Ip S. M. , Old D. C.
Published: January 2020

Tests for the presence of mannose-sensitive and mannose-resistant, eluting hemagglutinins and fimbriae were helpful in indicating whether biochemically atypical strains of the tribe
Escherichieae
might be escherichiae or shigellae.
Dermatophilus congolensis infections in wildlife in New York State
Authors: Salkin I F , Stone W B , Gordon M A
Published: January 2020

Dermatophilus congolensis infection is described for the first time in a woodchuck (Marmota monax) and a striped skunk (Mephitis mephitis). This actinomycete was also recovered from a raccoon (Procyon lotor) carcass which had been frozen for a year. Encrusted lesions or denuded patches or both were seen during necropsy on the head, thorax, abdomen, and legs of each carcass. D. congolensis was demonstrated in strained smears or paraffin sections or both from the three animals and was recovered from lesions induced in a rabbit's ear by inoculation with infected raccoon tissue. This expands to 59 the number of cases of dermatophilosis found in wildlife and domestic animals in New York State and vicinity.
Comparison of methods for detection of Mycoplasma pulmonis in experimentally and naturally infected rats
Authors: Davidson M K , Lindsey J R , Brown M B , Schoeb T R , Cassell G H
Published: January 2020

Isolation, indirect immunofluorescence, an enzyme-linked immunosorbent assay (ELISA), and histopathological examination of tissues for characteristic lesions were evaluated for their efficiency in detecting Mycoplasma pulmonis infection in rats. Whereas all of the methods were efficient in naturally infected Sprague-Dawley rats, none of the methods consistently detected infection in F344 rats experimentally infected with low doses of the organism. In the experimental infections, however, the success rate of any method was directly related (P less than 0.05) to increasing inoculum dose and time postinoculation. Collectively, the data indicated that isolation of M. pulmonis was the most efficient single detection method and the nasopharyngeal duct was the best single site to culture, although sampling of multiple sites within the respiratory tract increased the rate of isolating the organism. The ELISA was understandably the least sensitive method in the low-dose, experimentally infected rats because of the time required for development of a detectable serum antibody response. Although each of the four methods identified a high percentage of naturally infected rats, the ELISA was the most efficient method in these animals as it was uniformly positive. The use of combinations of methods was found to increase the rate of detection of M. pulmonis infection in both experimentally and naturally infected rats.
Time-motion and cost comparison study of micro-ID, API 20E, and conventional biochemical testing in identification of Enterobacteriaceae
Authors: Bale M J , Matsen J M
Published: January 2020

A total of 730 Enterobacteriaceae strains isolated from 567 cultures were evaluated by a rapid kit method (Micro-ID; General Diagnostics, Morris Plains, N.J.; 4 h), an overnight incubation kit method (API 20E; Analytab Products, Plainview, N.Y.), and conventional biochemical test methodology (mostly overnight incubation and some rapid methods) to compare the amount of laboratory effort required, timing, and cost parameters. We assessed the amount of technologist time expended, the time sequence of culture reporting to physicians, the number of isolates requiring repeat testing or additional biochemical testing, the number of cultures held due to the need for identification of other organisms, the cost of total work-up, etc. Cultures evaluated included urines, respiratory cultures, wound cultures, body fluids, genital cultures, and cultures from miscellaneous categories. A total of 64% of the Enterobacteriaceae strains processed by the Micro-ID method could be identified within 24 h of receipt of the specimens in the clinical laboratories, in contrast to the need for an additional day required by the API or conventional biochemical methods. The Micro-ID method also required less technologist time (4.5 min) for set-up and interpretation than did either the API method (6 min) or conventional methods (7 min). Total direct costs (June 1981) per organism identified were: Micro-ID, $4.30; API 20E, $4.96; conventional biochemicals with commercially prepared media, $5.66. An estimate of 80% technologist time efficiency was made in all procedures.
Identification of coagulase-positive Staphylococcus intermedius and Staphylococcus hyicus subsp. hyicus isolates from veterinary clinical specimens
Authors: Phillips W E , Kloos W E
Published: January 2020

Coagulase-positive isolates of Staphylococcus intermedius from dogs and coagulase-positive isolates of Staphylococcus hyicus subsp. hyicus from a pig and cows were identified initially by a simplified scheme which can be readily performed in a routine diagnostic laboratory. Characters tested in the scheme included coagulase activity, colony pigment, and aerobic acid production from maltose. The identity of these isolates was confirmed by deoxyribonucleic acid-deoxyribonucleic acid hybridization experiments. The strains of S. hyicus subsp. hyicus isolated from porcine sources were positive for protein A, whereas the strains recovered from bovine mastitic milk were negative for protein A.
Vibrio metschnikovii
Bacteremia in a Patient with Cholecystitis
Authors: Jean-Jacques W. , Rajashekaraiah K. R. , Farmer J. J. , Hickman F. W. , Morris J. Glenn , Kallick Charles A.
Published: January 2020

Vibrio metschnikovii
was isolated from the blood of an 82-year-old patient with peritonitis and an inflamed gallbladder. This is probably the first clinically significant isolate of this new vibrio.
Antibody response to Pseudomonas aeruginosa exoproducts in cancer patients
Authors: Crowe K E , Bass J A , Young V M , Straus D C
Published: January 2020

We analyzed sera from 28 patients with various types of malignancies for the occurrence of antibodies against exotoxin A of Pseudomonas aeruginosa and two Pseudomonas proteases. A total of 27 of these individuals were colonized or infected with P. aeruginosa at one time or another during the study, whereas the remaining patient was colonized with four non-P. aeruginosa species of Pseudomonas. Sera were obtained from several of these patients before P. aeruginosa colonization or infection of these individuals was detected, which provided an opportunity to evaluate their responsiveness to pseudomonal exoproducts as they acquired the organism. Exotoxin A was purified from culture supernatant fluids of strain PA-103, and the two proteases were purified from an isolate of strain JR3, a highly proteolytic strain originally recovered from the sputum of a cystic fibrosis patient. Antibodies to the exotoxin A and the two proteases were detected in these sera, and sera which contained relatively high antibody levels to exotoxin A afforded mice complete protection against lethal challenges with this substance. Statistical analyses showed that patients infected with P. aeruginosa had consistently higher antibody levels (P less than 0.005) to the exoproducts than patients who were colonized with this organism. Also, patients colonized with P. aeruginosa possessed significantly higher antibody levels (P less than 0.003) to these three exoproducts than uninfected, hospitalized patients. Parke-Davis type 1 was the strain most commonly isolated from these patients (46%), but colonization or infection due to this organism usually resulted in the production of low levels of antibody to Pseudomonas exoproducts. However, infections with Parke-David type 7 organisms were always associated with intermediate- and high-responder sera to exotoxin A. These results indicated that potentially toxic products were elaborated during the course of cancer-related colonization and infection with P. aeruginosa.
Enrichment medium and control system for isolation of Campylobacter fetus subsp. jejuni from stools
Authors: Chan F T , Mackenzie A M
Published: January 2020

Enrichment culture with a semisolid medium increased by 6% the isolation rate of Campylobacter fetus subsp. jejuni. The semisolid enrichment medium was also used successfully as a transport medium for Campylobacter isolates. A blood agar plate streaked with Pseudomonas aeruginosa, Clostridium perfringens, and a laboratory strain of Campylobacter was a good control system for the microaerophilic atmosphere. Good growth of all three organisms indicated satisfactory conditions for culturing Campylobacter.
Enzymatic activities of Legionella pneumophila and Legionella-like organisms
Authors: Nolte F S , Hollick G E , Robertson R G
Published: January 2020

The API ZYM system was used to investigate enzymatic activities of Legionella pneumophila and other Legionella-like organisms. Leucine aminopeptidase, alkaline and acid phosphatase, butyrate and caprylate esterase, and phosphoamidase activities were consistently detected in all strains tested. No evidence of myristate lipase, trypsin, chymotrypsin, or glycosidase activity was found.
Evaluation of the necessity for routine terminal subculturing of blood cultures negative by radiometric methods
Authors: Beckwith D G , Etowski D C
Published: January 2020

A retrospective review of 18,130 blood cultures performed with a BACTEC 225 indicated that terminal subculturing of bottles negative after 7 days of testing did not recover organisms which affected patient care or the length of patient hospitalization.
Evaluation of the CampyPak II gas generator system for isolation of Campylobacter fetus subsp. jejuni
Authors: Buck G E , Fojtasek C , Calvert K , Kelly M T
Published: January 2020

The CampyPak II (BBL Microbiology Systems, Cockeysville, Md.) method for isolating Campylobacter fetus subsp. jejuni was evaluated with clinical specimens by comparison with an evacuation replacement procedure. Of 757 specimens, 26 (3.4%) were positive for C. fetus subsp. jejuni. All 26 were recovered by both systems. No difference was found in the time necessary for isolation, except with one isolate that required 3 days for isolation with CampyPak II and 2 days with the established procedure. Although the average colony size was slightly smaller for colonies incubated in the CampyPak II system (P less than 0.02 by a paired t test), this is probably of no practical consequence. In terms of technical performance, therefore, the CampyPak II system appears to be an excellent alternative method for establishing microaerophilic conditions necessary for isolating C. fetus subsp. jejuni. The only difference between the two systems was the cost of using the CampyPak II envelopes. Excluding the initial equipment costs, CampyPak II envelopes cost approximately $0.74 per specimen, whereas the standard Torbal jar technique cost about $0.10 per specimen.
Identification of group D streptococci by SeroSTAT
Authors: Levchak M E , Ellner P D
Published: January 2020

Clinical isolates of group D streptococci presumptively identified by biochemical methods were grouped by latex agglutination using a commercially prepared reagent specifically sensitized with group D antiserum (SeroSTAT; Scott Laboratories, Inc., Fiskeville, R.I.). Streptococcus species tested included S. faecalis, S. faecium, S. durans, S. avium, S. bovis, and S. equinus. Colonies of the organism to be tested were picked from agar plates, emulsified in a drop of glycine-buffered saline on a slide, and mixed with a drop of the latex reagent. Macroscopic agglutination occurred within 60 s. A total of 115 isolates of group D streptococci were tested; 103 (89.6%) gave positive reactions with SeroSTAT. Twelve strains failed to react with the latex reagent; these 12 strains also gave negative results with group D antiserum when tested by the Lancefield method. Two of 14 group A streptococci also reacted with the SeroSTAT group D reagent; after trypsinization, the cross-reaction was eliminated. Group B streptococci, viridans group streptococci, anaerobic streptococci, and staphylococci all gave negative reactions with the SeroSTAT reagent. The SeroSTAT reagent is a useful diagnostic tool for the prompt identification of enterococcal and non-enterococcal group D streptococci.
Determination of immune complexes in sera from dogs with various diseases by mastocytoma cell assay
Authors: Targowski S
Published: January 2020

Canine immunoglobulin G complexed with particulate or soluble antigen can bind to the Fc receptors on the mastocytoma cells. Attachment of immune complexes composed of immunoglobulin G and soluble antigen (ovalbumin) to mastocytoma cells was detected by an inhibition of rosette formation with indicator cells (sensitized sheep erythrocytes). Therefore, canine circulating immune complexes may also attach to mastocytoma cells and inhibit rosette formation (mastocytoma cell assay). Sera from 326 dogs with various diseases and from 50 clinically normal dogs were assayed for immune complexes. The incidence of immune complexes in sera from normal dogs was 6% as compared with 25% in dogs with various diseases. The immune complexes were demonstrated in 37% of sera from dogs with various neoplastic diseases, 40% of sera from dogs with diabetes, 24% of sera from dogs with hypothyroidism, 50% of sera from dogs with mycotic disease, 75% of sera from dogs with arthritis, 38% of sera from dogs with kidney disorders, 40% of sera from dogs with neurological diseases, 45% of sera from dogs with various parasitic diseases, and 27% of sera from dogs with liver disorders. Only 19% of sera from dogs admitted to the hospital for various surgeries gave positive results. The incidence of the positive sera from dogs with various diseases is discussed in regard to their counterparts of human diseases.
Effect of gaseous conditions on isolation and growth of Ureaplasma urealyticum on agar
Authors: Robertson J A
Published: January 2020

Five laboratory-adapted strains of Ureaplasma urealyticum showed comparable colony counts when incubated in eight gaseous environments including air, in different concentrations of carbon dioxide, and under anaerobiosis. For primary isolation on genital mycoplasma agar 95% N2-5% CO2 gave 100% correlation with growth in bromothymol blue broth, whereas certain strains failed to grow on agar incubated in air, the TABCO2 system (Lab-Tek Division, Miles Laboratories, Ltd., Rexdale, Canada), 100% CO2, or under anaerobiosis.
Production and characterization of a hybridoma-derived antibody to Blastomyces dermatitidis
Authors: Young K D , Larsh H W
Published: January 2020

A hybridoma cell line was isolated which produced monoclonal antibody to one protein component of a yeast-phase cytoplasmic antigenic complex of Blastomyces dermatitidis. The immunoglobulin M antibody product was characterized by immunodiffusion, autoradiography of polyacrylamide gels, and cellulose acetate electrophoresis. By attaching the antibody to an affinity gel, one major protein band was identified by polyacrylamide gel electrophoresis as the antigen for which the antibody was specific.
Enzyme-linked immunosorbent assay for detection of Salmonella typhi Vi antigen in urine from typhoid patients
Authors: Barrett T J , Snyder J D , Blake P A , Feeley J C
Published: January 2020

Because typhoid fever continues to be a major cause of illness in many developing countries, there is a clear need for a sensitive and specific test that will permit rapid laboratory diagnosis of the disease. An enzyme-linked immunosorbent assay (ELISA) has recently been developed and tested, both in the laboratory and in a clinical situation, for its ability to detect Vi antigen in urine. The ELISA was capable of detecting as little as 1 ng of purified Vi antigen per ml in urine, compared with 100 ng/ml detectable by a previously tested coagglutination method. It could also detect antigen in urine diluted as much as 1:1,024 in normal urine. In tests of urine specimens from six stool culture-positive persons in a small typhoid outbreak in the United States, the ELISA detected antigen in specimens from four of the six patients. The ELISA also proved to be specific, giving no false-positive results for specimens from 50 persons who did not have typhoid fever. The apparent high sensitivity and specificity of this ELISA make it a promising test for rapid diagnosis of typhoid fever.
Improved immunofluorescence antigens for detection of immunoglobulin M antibodies to Epstein-Barr viral capsid antigen and antibodies to Epstein-Barr virus nuclear antigen
Authors: Gallo D , Walen K H , Riggs J L
Published: January 2020

Epstein-Barr viral capsid antigen and nuclear antigen produced by modified procedures were evaluated for use in measuring viral capsid antigen immunoglobulin M and Epstein-Barr virus nuclear antigen antibody responses in sera from patients with suspected Epstein-Barr virus infections. Viral capsid antigen production was stimulated with a phorbol ester, and the Epstein-Barr virus nuclear antigen cells were fixed in suspension to eliminate loss of antigen during the drying process. Both preparations proved to be sensitive and reliable.
Evaluation of the Antibiotic Removal Device
Authors: Appleman M D , Swinney R S , Heseltine P N
Published: January 2020

A new system called an Antibiotic Removal Device (ARD), used for the removal of residual antibiotics in blood specimens, was studied in 300 patients diagnosed as clinically septic despite antimicrobial therapy. Blood specimens from these patients were processed with and without the ARD into aerobic and anaerobic media. There were 53 patients who had one or more positive blood culture bottles, for a total of 109 positive blood culture bottles. Of these 109 bottles, 33.9% were positive with ARD-processed blood in aerobic media, 24.8% with ARD-processed blood in anaerobic media, 22.9% with conventionally processed blood in aerobic media, and 18.3% with conventionally processed specimens in anaerobic media. After 6 h of incubation, 14 bottles with blood processed by the ARD and 9 bottles processed conventionally were positive; after 24 h, 48 bottles with blood processed by the ARD and 31 bottles processed conventionally were positive. The same organisms were isolated from both ARD- and conventionally processed blood, with four exceptions. The ARD system, when compared with a conventional system of blood specimen processing, significantly increased the detection of bacteremia and decreased the time required for its detection in patients clinically septic despite ongoing antimicrobial therapy.
Evaluation of the AutoMicrobic system for identification of glucose-nonfermenting gram-negative rods
Authors: Smith S M , Cundy K R , Gilardi G L , Wong W
Published: January 2020

With the Enterobacteriaceae Plus Biochemical Card, the AutoMicrobic system (Vitek Systems, Inc., Hazelwood, Mo.) is purported to identify members of the family Enterobacteriaceae and seven species of glucose-nonfermenting gram-negative rods: Pseudomonas aeruginosa, P. fluorescens, P. putida, P. maltophilia, P. cepacia, and Acinetobacter calcoaceticus (saccharolytic and non-saccharolytic). The latter capability was examined in this investigation. Of 410 glucose-nonfermenting rods included in the identification profile, the AutoMicrobic system correctly identified 366 (89.3%). Of 62 glucose-nonfermenting organisms not included in the identification profile, 41 (66%) were correctly reported as "unidentified organism." The usefulness of the AutoMicrobic system-generated identification probability in establishing criteria for acceptance of identification is discussed.
Porcine pararotavirus: detection, differentiation from rotavirus, and pathogenesis in gnotobiotic pigs
Authors: Bohl E H , Saif L J , Theil K W , Agnes A G , Cross R F
Published: January 2020

Some characteristics of a newly recognized porcine enteric virus are described. Tentatively, the virus was referred to as porcine pararotavirus (PaRV) because it resembled rotaviruses in respect to size, morphology, and tropism for villous enterocytes of the small intestine. However, it was antigenically distinct from porcine, human, and bovine rotaviruses and reoviruses 1, 2, and 3, and the electrophoretic migration pattern of PaRV double-stranded RNA was distinct from the electrophoretic migration patterns of the rotaviral and reoviral genomes. By passage in gnotobiotic pigs, PaRV was isolated from two suckling diarrheic pigs originating from two herds. After oral exposure of gnotobiotic pigs, villous enterocytes of the small intestines became infected as judged by immunofluorescence, resulting in villous atrophy and diarrhea. Mortality was high when gnotobiotic pigs less than 5 days old were infected. The C strain of this virus was serially passed 10 times in gnotobiotic pigs, and electron microscopy, immunofluorescence, and serological tests indicated no extraneous agents. The virus was serially passed five times in cell cultures which contained pancreatin in the medium, but replication was negligible or absent, as the number of immunofluorescent cells decreased with each passage. Since rotaviral infections are frequently diagnosed by direct electron microscopy of fecal specimens, the presence of other morphologically similar viruses, such as PaRV, should be considered. The use of immune electron microscopy is suggested as a means of helping recognize this situation.
Novel approach to transfer and preparation of replicate mycological cultures
Authors: Swanson G C , Prince J T
Published: January 2020

A technique with the use of miniature brushes and single inoculum onto multiple slants or plates is described. The technique drastically reduced the time usually required for the preparation of replicate cultures in the teaching of mycology without the sacrifice of macroscopic or microscopic cultural characteristics.
Heated versus unheated sera in the hemagglutination treponemal test for syphilis
Authors: Hambie E A , Larsen S A , Perryman M W , Pettit D E , Feeley J C
Published: January 2020

Sera (920) were tested to evaluate the use of heated versus unheated sera in the hemagglutination treponemal test for syphilis. The heated and unheated samples were tested on the same day with the hemagglutination treponemal test for syphilis kit according to the manufacturer's protocol. Agreement of results between the heated and unheated sera was 99.2%. The reading pattern of agglutination was clearer and more distinct with heated sera; therefore, based solely on our preference for the reading patterns, we suggest that heated sera be used in the hemagglutination treponemal test for syphilis.
Simple, transparent medium for study of legionellae
Authors: Johnson S R , Schalla W O , Wong K H , Perkins G H
Published: January 2020

A simple medium containing yeast extract, phosphate, and hemin as major components has been formulated for the study of legionellae. The medium supports the growth of a wide range of clinical and environmental isolates, with plating efficiencies comparable to those of charcoal-yeast extract agar. In addition, it does not contain activated charcoal or other components which may be associated with adverse reactions in humans. The medium is buffered and transparent; therefore, it would be suitable for genetic studies, production of biological reagents such as antigens and skin tests for human use, and antibiotic susceptibility assays.
Type-specific antigens of Neisseria meningitidis serogroup A
Authors: Di Ninno V L , Chenier V K , Bhatti A R , White L A
Published: January 2020

By using Ouchterlony immunodiffusion techniques, we defined three unique antigenic determinants (A1, A2, and A3) among six isolates of Neisseria meningitidis serogroup A. Two of these determinants (A2 and A3) were identified as lipopolysaccharides in nature and were not found to occur on the serogroup B strains of N. meningitidis that we examined. The A1 determinant commonly occurred among the serogroup B strains and was identified as a protein determinant.
Carpeting in hospitals: an epidemiological evaluation
Authors: Anderson R L , Mackel D C , Stoler B S , Mallison G F
Published: January 2020

Epidemiological and microbiological studies were conducted in a hospital room with carpet (CR) and in one with carpet (NCR). Microbiological profiles were determined with specimens obtained from patients admitted to these rooms. Patient records were reviewed to note infection status and other case identities. Eleven-millimeter cylindrical core samples of carpet were obtained, and swab template techniques were used on the bare floor for subsequent enumeration and identification of contaminating microorganisms. In each sampling period, higher microbial counts per square inch (1 in(2) = ca 6.452 cm(2)) were measured for the carpet than for the bare floor. Recovery rates of Enterobacter spp., Klebsiella pneumoniae, and Escherichia coli were higher from carpet samples than from bare floor samples. Typable organisms (such as E. coli, Pseudomonas aeruginosa, K. pneumoniae, and Staphylococcus aureus) obtained from patients were also more frequently recovered from the carpet than from the bare flooring. Patients who stayed in the CR were shown to be colonized with the same types of organisms as those initially recovered from the carpet. However, no statistically significant differences were found in patients in the CR versus NCR in colonization with all typable and nontypable organisms first found on the floor. Disease in patients was found not to be associated with organisms found as contaminants of the carpet or the bare floor. Air above carpeting contained more consistent concentrations of organisms than air above the bare flooring.
Evaluation of a commercial latex agglutination test for identification of Staphylococcus aureus
Authors: Doern G V
Published: January 2020

A new, commercially available latex agglutination test (SeroSTAT Staph; Scott Laboratories, Inc., Fiskeville, R.I.) was compared with the tube coagulase and slide coagulase tests as means for identifying Staphylococcus aureus. Of 160 clinical isolates of S. aureus, 159 (99.4%) yielded positive results with the latex agglutination test. Negative latex agglutination test results were obtained with 266 of 267 clinical isolates of Micrococcus spp. and staphylococcal species other than S. aureus (99.6%). The latex agglutination test was found to be a rapid, technically nondemanding method for identifying S. aureus. It was as accurate as the tube coagulase test and more accurate than the slide coagulase test.
Identification of problem Neisseria gonorrhoeae cultures by standard and experimental tests
Authors: Arko R J , Finley-Price K G , Wong K H , Johnson S R , Reising G
Published: January 2020

Standard and experimental tests were used by a reference diagnostic laboratory to determine the identity of 182 "suspected" Neisseria gonorrhoeae isolates submitted by state health departments because of inconclusive laboratory results. More than 97% of these cultures were subsequently identified by a rapid microcarbohydrate test in conjunction with confirmatory immunological procedures. The experimental rapid slide agglutination test using rough-lipopolysaccharide antibody, the Phadebact co-agglutination test, and fluorescent antibody test identified 49.3 to 94.1% of these cultures. Because of frequent problems with carbohydrate utilization, Neisseria meningitidis and Branhamella catarrhalis were the two microorganisms most often confused with N. gonorrhoeae by submitting laboratories.
Natural and experimental infection of woodchucks with woodchuck hepatitis virus, as measured by new, specific assays for woodchuck surface antigen and antibody
Authors: Wong D C , Shih J W , Purcell R H , Gerin J L , London W T
Published: January 2020

Solid-phase radioimmunoassays for woodchuck hepatitis virus (WHV) surface antigen (WHsAg) and antibody to it (anti-WHs) were developed. The test for WHsAg could detect as little as 10 ng/ml. In both tests it was necessary to employ radiolabeled WHsAg instead of anti-WHs as the probe because the latter appeared to be labile to the conditions of labeling. The tests were used to characterize naturally acquired and experimental WHV infections of woodchucks. Forty-three of 72 wild-caught woodchucks had serological evidence of WHV infections; 16 of these resulted in chronic infection, and the remainder were self-limiting. All chronically infected animals were positive for WHsAg and DNA polymerase activity. During 3 years of observation, 11 of the 16 WHsAg-positive animals and 3 of the 27 anti-WHs-positive animals, but none of the 21 uninfected animals developed hepatocellular carcinoma. Seroconversion, possibly resulting from infection with WHV, was documented in a chimpanzee inoculated with WHV. An immune adherence hemagglutination test for WHsAg was also developed by using anti-WHs of chimpanzee origin as a reagent, but the test was not useful for detecting anti-WHs of woodchuck origin because of the lability of the latter.
Effect of various analgesics and lubricants on isolation of Chlamydia trachomatis and Neisseria gonorrhoeae
Authors: Osborn M F , Johnson A P
Published: January 2020

Five preparations used as analgesics or lubricants in surgical, obstetrical, gynecological, and investigative procedures were tested for their effect on the isolation of Chlamydia trachomatis. Three lignocaine preparations and a lubricating jelly containing 2% phenol were inhibitory to chlamydiae. In contrast, K-Y lubricating jelly was relatively nontoxic to chlamydiae. Since K-Y jelly also had only slight toxic activity against gonococci, it is recommended for the lubrication of instruments which need to be used for the efficient isolation of these microorganisms.
Differentiation of coagulase-positive and coagulase-negative staphylococci by lectins and plant agglutinins
Authors: Davidson S K , Keller K F , Doyle R J
Published: January 2020

The screening of staphylococci with a panel of 14 lectins and extracts demonstrating lectin-like activity led to the development of a rapid agglutination slide test for the differentiation of certain coagulase-negative staphylococci and human strains of Staphylococcus aureus. The coagulase-negative staphylococci were agglutinated by agglutinins from Mangifera indica, Triticum vulgaris, and crude Limulus polyphemus. The test is rapid, requiring only 5 to 15 min to identify an unknown strain of staphylococci, as opposed to the 4 to 16 h required to perform the conventional tube coagulase test.
Use of a hemadsorption technique to evaluate the stability of the hemagglutination reaction of Escherichia coli cultures possessing human colonization factor antigens
Authors: Sedlock D M , Bartus H F , Zajac I , Actor P
Published: January 2020

Two strains of Escherichia coli, producing different colonization factor antigens (CFA), were monitored for the population density of CFA-producing bacteria after repeated subculture. The production of CFA was estimated by flooding agar plates containing isolated colonies with suspensions of human or bovine erythrocytes. The erythrocytes were suspended in a low-ionic-strength buffer and were fixed to CFA-positive colonies with a 1.0% tannic acid solution. Strain H-10407, possessing CFA/I fimbriae, showed a rapid loss of the hemagglutinin when subcultured, whereas strain CL-9699, producing CFA/II, was very stable. By using the hemadsorption assay, we could rapidly and easily distinguish CFA- positive colonies from the CFA-negative variants. A survey of additional E. coli strains demonstrated the utility and specificity of the hemadsorption technique used.
Controlled evaluation of the volume of blood cultured in detection of bacteremia and fungemia
Authors: Tenney J H , Reller L B , Mirrett S , Wang W L , Weinstein M P
Published: January 2020

To evaluate the role of the volume of blood cultured in the detection of clinically important bacteremia and fungemia in adults, we evaluated the yield and speed of detection of microorganisms from 5,317 paired 2- and 5-ml samples of blood. The same kind of medium (supplemented peptone broth with 0.03% sodium polyanetholsulfonate) and atmosphere of incubation (open venting units) were used for all blood cultures. Only adequately filled (less than or equal to 80% of stated volume) sets (20-ml tube and 50-ml bottle) were compared statistically. Significantly more bacteria (p less than 0.01), Pseudomonas spp. In particular (P less than 0.05), were isolated from the 5-ml samples of blood. We conclude that the volume of blood cultured is a critical factor in the detection of septicemia. Consequently, valid evaluation of other factors influencing the detection of septicemia must be based on comparisons in which equal volumes of blood are cultured.
Spot indole test: evaluation of four reagents
Authors: Miller J M , Wright J W
Published: January 2020

Kovacs indole reagent, p-dimethylaminobenzaldehyde, Ehrlich indole reagent and p-dimethylaminocinnamaldehyde were used as spot indole reagents to test 359 strains of gram-negative rods growing on 5% sheep blood agar, Trypticase soy agar (BBL Microbiology Systems), and MacConkey agar. The p-dimethylaminocinnamaldehyde reagent was the most sensitive of those tested and provided results that were easiest to interpret. The p-dimethylaminocinnamaldehyde reagent was able to detect providencia alcalifaciens indole production because of the red-violet color unique to that organism. All reagents tested were accurate in detecting indole produced by members of the Enterobacteriaceae family, with the exception of P. alcalifaciens.
Isolation and cultivation of Haemophilus ducreyi
Authors: Oberhofer T R , Back A E
Published: January 2020

A useful method for isolating and recognizing Haemophilus ducreyi from chancres and buboes of male patients is presented. A total of 41 clinical isolates of H. ducreyi were recovered from 33 patients over an 8-year period, and the experience with the 15 most recent isolates is presented in detail. Chocolate agar supplemented with 1% Iso VitaleX and 5% sheep blood agar were prepared, using Trypticase soy and Mueller-Hinton Agar bases, and incubation conditions included ambient, capneic, and anaerobic environments. Mueller-Hinton agar was clearly superior over Trypticase soy agar for isolation of H. ducreyi, although there was little difference between 5% sheep blood and supplemented chocolate agar. Growth in ambient air and under anaerobiasis was poor or lacking, whereas growth in 5 to 7% CO2 was good to luxuriant. Heat-inactivated and fresh (unheated)human blood clot tubes also were used for selective isolation. Although the rates of isolation from the two types of clot tube were not significantly different, unheated clot tubes were superior to heated clot tubes because of reduced level of contaminants. Gram stain characteristics taken from blood clot tubes and solid media, cellular and colonial morphology of the bacilli, and lack of oxidase, catalase, and biochemical activity except nitrate reductase were determinant factors. The results of this study demonstrated that successful isolation of H. ducreyi can be achieved with a minimal amount of resources and expertise.
Use of the automicrobic and enteric-tek systems for identification of Enterobacteriaceae
Authors: Goldstein J , Guarneri J J , Della-Latta P , Scherer J
Published: January 2020

Studies were performed to evaluate the use of the Enteric-Tek (ET) and the AutoMicrobic system (AMS) for the identification of 201 Enterobacteriaceae freshly isolated from clinical specimens. All test systems were inoculated simultaneously from the same MacConkey agar plate. Organisms were also identified with conventional media. Identifications with the ET and AMS agreed with those made with conventional biochemicals 97% of the time. At a 95% confidence level, the ET was able to identify 75% of the isolates within 18 h without the aid of additional biochemical tests; the AMS identified 92% in 8 h. Technologist time needed for identifications made with the AMS was reduced approximately 57% compared with the ET. In all instances in which the AMS identification disagreed with the conventional, the ET identified the organism correctly. Similarly, organisms misidentified by the ET were correctly identified by the AMS. The data suggest that the AMS and ET identify clinical isolates with comparable accuracy; however, the AMS offers a significant savings in time.
Comparative measurement of equine influenza virus antibodies in horse sera by single radial hemolysis, neutralization, and hemagglutination inhibition tests
Authors: Yamagishi H , Nagamine T , Shimoda K , Ide S , Igarashi Y , Yoshioka I , Matumoto M
Published: January 2020

Single radial hemolysis (SRH), neutralization (NT), and hemagglutination inhibition (HI) tests were carried out on sera from horses immunized against the Prague and Miami strains of equine influenza virus. The HI and NT tests demonstrated good sensitivity; the sensitivity of the SRH test was somewhat lower. The NT titers of individual sera were correlated very closely with the HI titers, although the NT titers were higher. SRH zone diameters of individual sera also showed significant correlation with the NT and NI titers. The SRH test appears to be suitable for large-scale serological surveys and offers the advantages of rapidity and simplicity.
Electrophoretic study of Clostridium species
Authors: Cato E P , Hash D E , Holdeman L V , Moore W E
Published: January 2020

Polyacrylamide gel electrophoretic analysis of soluble cellular proteins (without sodium dodecyl sulfate) of 70 Clostridium species indicated that the procedure was readily applicable to the differentiation of species in the genus. The protein patterns correlated well with the available DNA homology data and with most accepted differential tests. Results indicated that several earlier names for species were synonyms of those of accepted species and that two accepted species may be synonymous.
Aniline blue-containing buffered charcoal-yeast extract medium for presumptive identification of Legionella species
Authors: Holmes R L
Published: January 2020

By utilizing buffered charcoal-yeast extract medium containing 0.01% aniline blue in conjunction with a long-wave UV light, the differentiation of five species of Legionella was facilitated. L. pneumophila, when grown on this medium, did not absorb the aniline blue dye; however, L. micdadei, L. dumoffii, L. bozemanii, and L. gormanii absorbed the dye in varying amounts and produced colonies of various shades of blue.
Clinical significance of viridans streptococci isolated from blood cultures
Authors: Swenson F J , Rubin S J
Published: January 2020

The clinical significance of viridans streptococci isolated from the blood cultures of 86 patients was determined. Isolates that were significant or suggestive of infection represented only 21% of the cases. Among 54 isolates for which the species was known, Streptococcus sanguis II was the most common. However, a significant association between species and clinical significance was not found.
Nosocomial outbreak of nitrate-negative Serratia marcescens infections
Authors: Geiseler P J , Harris B , Andersen B R
Published: January 2020

Bacteremia due to multiply-antibiotic-resistant Serratia marcescens occurred within 1 week in four patients who were in adjacent beds in an intensive care unit. The strains were serotyped as O14:H12 and were nitrate negative. This unusual biochemical marker was useful in the investigation of the outbreak.
Rapid micromethod for preparation of enzyme-antibody conjugates
Authors: Parkinson A J , Scott E N , Muchmore H G
Published: January 2020

A rapid method is described for labeling antibody with alkaline phosphatase by one-step glutaraldehyde linkage. The method involves the centrifugation of a small volume of an enzyme and antibody mixture through a minicolumn packed with hydrated Sephadex. This procedure rapidly removes ammonium sulfate and glutaraldehyde from the enzyme-antibody mixture and results in the efficient recovery of conjugated antibody without significant dilution.
Identification of DNA viruses by membrane filter hybridization
Authors: Stålhandske P , Pettersson U
Published: January 2020

The use of membrane filter hybridization for the identification of DNA viruses is described. We designed and used a procedure for identification of herpes simplex virus. This method can discriminate between herpes simplex virus types 1 and 2 in a simple way.
Comparison of fluorescent and colorigenic substrates for enzyme immunoassays
Authors: Yolken R H , Leister F J
Published: January 2020

A variety of substrates can be employed in enzyme immunoassays (EIAs) for the measurement of enzyme-labeled immunoreactants. We compared the sensitivities of a fluorescent and a colorigenic substrate in an EIA system for the measurement of Haemophilus influenza purified polyribose phosphate. After a 10-min substrate incubation, the EIA in which the fluorescent substrate was used could detect 10 pg of polyribose phosphate per ml, whereas the EIA in which the colorigenic substrate was used required the addition of 640 pg of polyribose phosphate per ml to generate a positive reading. However, the use of longer substrate incubation periods led to an increase in sensitivity of the colorigenic EIA. After an incubation period of 240 min, the sensitivity was equal to that of the EIA in which the fluorescent substrate was used. These results suggest that the ultimate limit of sensitivity of EIA systems is determined by the nature of the antigen-antibody reactions. However, the use of high-energy substrates in EIA systems can allow for the attainment of maximal sensitivity after short enzyme-substrate incubation periods.
Cefoperazone disk diffusion susceptibility test: confirmation of the tentative interpretive criteria, Pseudomonas aeruginosa cross-resistance, and determination of quality control performance limits
Authors: Jones R N , Gavan T L , Barry A L , Thornsberry C , Gibbs D L
Published: January 2020

Cefoperazone disk diffusion test and minimum inhibitory concentration comparison studies were performed on 421 recent bacterial isolates, using 30- and 75-micrograms commercially prepared disks. Acceptable correlation coefficients (-0.82 to -0.86) and very major (false-susceptible) interpretive error rates (less than 1%) were obtained with both disk concentrations. The interpretive criteria for both disks were identical. Using the preferred 75-micrograms disk, the Thornsberry et al. criteria (J. Clin. Microbiol. 15:769-776, 1982) of greater than or equal to 18 mm = susceptible (less than or equal to 32 micrograms/ml) and less than or equal to 14 mm = resistant (greater than 64 micrograms/ml) resulted in only 5.5% of strains having indeterminate-range zone diameters; the 30-micrograms disk had 6.9% of strains with indeterminate zone diameters. The 75-micrograms disk, excluding the testing of enterococci, minimized the very major and other interpretive errors to less than 5%. Larger zone diameters will contribute few technical problems with either disk concentration. Data from 1,320 zone diameters submitted for each quality control strain indicated no significant (P greater than 0.05) difference between disks made by the three major manufacturers, and consistent results were obtained within each laboratory with numerous lots of Mueller-Hinton agar (except for one manufacturer). Individual daily test and accuracy quality control ranges were calculated from clinical investigator laboratory data at 16 hospitals based on mean zone sizes and from an additional 8 laboratories with both mean and median calculations. The quality control data were nearly identical, and ranges calculated by the two methods were very similar. Susceptibility tests of Pseudomonas aeruginosa indicate that the cefoperazone disk or minimum inhibitory concentration test would accurately predict P. aeruginosa susceptibility test results for other pseudomonas-active cephalosporins (cefsulodin and ceftazidime), thus producing no very major interpretive errors.
Analysis of the disagreement between automated bioluminescence-based and culture methods for detecting significant bacteriuria, with proposals for standardizing evaluations of bacteriuria detection methods
Authors: Nichols W W , Curtis G D , Johnston H H
Published: January 2020

A fully automated method for detecting significant bacteriuria is described which uses firefly luciferin and luciferase to detect bacterial ATP in urine. The automated method was calibrated and evaluated, using 308 urine specimens, against two reference culture methods. We obtained a specificity of 0.79 and sensitivity of 0.75 using a quantitative pour plate reference test and a specificity of 0.79 and a sensitivity of 0.90 using a semiquantitative standard loop reference test. The majority of specimens negative by the automated test but positive by the pour plate reference test were specimens which grew several bacterial species. We suggest that such disagreement was most likely for urine containing around 10(5) colony-forming units per ml (the culture threshold of positivity) and that these specimens were ones contaminated by urethral or vaginal flora. We propose standard procedures for calibrating and evaluating rapid or automated methods for the detection of significant bacteriuria and have analyzed our results using these procedures. We recommend that identical analyses should be reported for other evaluations of bacteriuria detection methods.