bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of BIOGRAM and commercial microdilution antimicrobial susceptibility test systems
Authors: Sautter R L , Denys G A
Published: January 2020

The BIOGRAM (Difco Laboratories, Detroit, Mich.) system, which is designed to calculate MICs from disk diffusion zone diameters, was compared with two commercial microdilution antimicrobial susceptibility systems. A total of 111 clinical isolates were evaluated with each test system. Six additional isolates were tested in a comparison between BIOGRAM and Sceptor (Johnston Laboratories, Inc. Towson, Md.) systems. BIOGRAM demonstrated an overall correlation with the Sceptor microdilution method of 95.7% for 1,287 organism-antimicrobial susceptibility combinations. The BIOGRAM and UniScept (Analytab Products, Inc., Plainview, N.Y.) systems were in agreement in 90.3% of 1,048 organism-antimicrobial susceptibility combinations tested. All methicillin-resistant staphylococci were detected by the standard disk agar diffusion method used with the BIOGRAM system. The BIOGRAM system provides an acceptable alternative to these commercial systems for the determination of quantitative susceptibility.
Genomic variation of adenovirus type 5 isolates recovered from bone marrow transplant recipients
Authors: Webb D H , Shields A F , Fife K H
Published: January 2020

We characterized the genomic variation of adenovirus type 5 isolates recovered from bone marrow transplant recipients in Seattle between 1976 and 1982. By restriction endonuclease analysis, we identified three new adenovirus genomic variants, each associated with a single invasive adenovirus infection. In addition, we were able to obtain suggestive evidence for a nosocomial spread of a particular group of isolates within this population. This study demonstrates that the technique of restriction endonuclease analysis is an important epidemiological tool for investigating viral infections.
Comparison of TestPack Strep A test kit with culture technique for detection of group A streptococci
Authors: Schwabe L D , Small M T , Randall E L
Published: January 2020

Results obtained with Abbott Laboratories TestPack Strep A (TPSA), a 7-min enzyme immunoassay method, were compared with culture results to measure the ability of this assay to detect group A streptococci directly from 365 throat swabs. Our study demonstrated a sensitivity of 90.0% and a specificity of 97.4% for TPSA compared with cultures incubated for 48 h. The positive and negative predictive values of this assay versus the culture method were 92.8 and 96.3%, respectively. If specimens that provided fewer than 10 colonies per plate of group A streptococci are eliminated from the data, the sensitivity is increased to 95.6%. Additionally, 10 group A and 40 non-group A streptococcal isolates were tested directly with TPSA for the ability to distinguish group A from non-group A streptococci. All 50 isolates were correctly identified (100% accuracy). TPSA is a rapid, accurate, and easy-to-interpret method for detection and confirmation of group A streptococcal antigen directly from throat swabs and pure culture isolates.
Effect of agitation and frequent subculturing on recovery of aerobic and facultative pathogens by Roche Septi-Chek and BACTEC blood culture systems
Authors: Kim M J , Gottschall R L , Schwabe L D , Randall E L
Published: January 2020

The positivity rate and time to recovery of pathogens were compared in Roche Septi-Chek (RSC-TSB) and BACTEC radiometric systems on 3,539 paired blood cultures. Both systems were steadily agitated, with frequent subculturing or processing of the RSC-TSB agar slides and BACTEC bottles, respectively, during the first 24 h of incubation. The RSC-TSB system recovered 249 pathogens (7.0% positivity rate), compared with 234 (6.6% positivity rate) isolates recovered from BACTEC. For the most common isolates, Staphylococcus aureus and the Enterobacteriaceae, the median time to detection was 15.8 h for BACTEC and 18.6 h for the RSC-TSB system. No statistically significant difference was observed in recovery of organisms from the two systems, except for S. aureus (P less than 0.05). In the RSC-TSB system, 42% of S. aureus, 58% of the Enterobacteriaceae, and 45% of Pseudomonas aeruginosa isolates had sufficient growth on the agar slant to allow performance of rapid standardized identification and susceptibility studies. In comparison with other studies using static incubation, it appears that agitation and frequent subculturing of the RSC-TSB system during the first 24 h of incubation decreased the time to detection for the majority of significant blood culture isolates.
Detection and quantitation of toxic shock syndrome toxin 1 in vitro and in vivo by noncompetitive enzyme-linked immunosorbent assay
Authors: Rosten P M , Bartlett K H , Chow A W
Published: January 2020

Toxic shock syndrome toxin 1 (TSST-1), an exotoxin produced by many Staphylococcus aureus strains, is implicated as the prime causal agent of toxic shock syndrome (TSS). A sensitive and specific noncompetitive enzyme-linked immunosorbent assay (ELISA) capable of detecting TSST-1 at concentrations from 0.5 to 16 ng/ml was developed. This assay did not detect other staphylococcal enterotoxins including A, B, C1, C2, C3, D, and E. Possible interactions with protein A were readily eliminated by pretreatment of test samples with 10% normal rabbit serum. The assay was adapted for rapid screening of TSST-1 production by S. aureus isolates in culture supernatants in vitro and for detection of TSST-1 in vaginal washings of TSS patients and healthy controls in vivo. All 35 S. aureus isolates confirmed to be TSST positive by Ouchterlony immunodiffusion and 59 of 60 isolates confirmed to be TSST-1 negative gave concordant results by ELISA. Interestingly, toxigenic S. aureus strains isolated from TSS patients quantitatively produced significantly more TSST-1 in vitro compared with toxigenic control strains (P less than 0.05, Mann-Whitney rank sum test). TSST-1 could be detected by ELISA in three of four vaginal washings collected within 3 days of hospitalization from three women with acute menstrual TSS, compared with 0 of 17 washings from nine TSS patients hospitalized longer than 3 days (P = 0.003, Fisher's exact test) and 1 of 15 washings from 14 healthy control women (P = 0.016). This noncompetitive ELISA should be particularly useful for rapid screening of TSST-1 production by S. aureus isolates, for the purification and biochemical characterization of TSST-1, and for human and animal studies of the pathogenesis of TSS.
Comparison of TRACK XI fluorometric immunoassay system with other serologic tests for detection of serum antibody to Brucella abortus in cattle
Authors: Hall S M , Confer A W
Published: January 2020

The TRACK XI system (TRACK) is a commercially available fluorometric immunoassay system that has been adapted for the detection of antibodies to several antigens in various animal species. Serum antibodies to Brucella abortus were detected by TRACK and compared with those obtained from two primary binding assays (a fluorometric immunoassay [FIAX] and an enzyme-linked immunosorbent assay) and three standard serological tests (complement fixation, Rivanol precipitation, and CARD tests). A total of 298 serum samples were tested by each serological test. Of these serum samples, 134 were negative controls, 43 were from cattle 1 month after vaccination with B. abortus 19, and 121 were from cattle 10 to 12 weeks after a midgestational, intraconjunctival challenge with B. abortus 2308. The results of this study indicated that TRACK is both reproducible and accurate. The results compared favorably with those of other serological methods. TRACK is more rapid than either the enzyme-linked immunosorbent assay or FIAX system. TRACK was the most sensitive (96.3%) test, with a specificity of 100%.
Efficiency of immunofluorescence for rapid detection of common respiratory viruses
Authors: Ray C G , Minnich L L
Published: January 2020

Rapid immunofluorescence (FA) methods for the detection of common respiratory viruses were compared with culture results over a 3-year period to assess the relative efficiency of FA in a clinical laboratory setting. For respiratory syncytial virus, efficiencies were high (sensitivity, 90 to 95%; specificity, 92 to 95%). The sensitivity of FA for detection of parainfluenza virus type 1, parainfluenza virus type 3, influenza A virus, and adenoviruses ranged from 28 to 63%, but specificities for these viruses were uniformly 98 to 100%. The observations form a basis for consideration of selective reduction of routine culture procedures for specimens with initial positive rapid FA results; however, the possibility of dual viral infection in some situations must also be considered.
Improved method for recovery of peritonitis-causing microorganisms from peritoneal dialysate
Authors: Ryan S , Fessia S
Published: January 2020

The efficacy of recovery of peritonitis-causing microorganisms from peritoneal dialysate fluid by using the Septi-Chek blood culture system (Roche Diagnostics, Div. Hoffmann-La Roche Inc., Nutley, N.J.) was compared with those of other conventional techniques, such as the 20-ml culture and the filtration methods. The recovery of microorganisms by using the Septi-Chek system was found to be as effective as the filtration of 250 ml of dialysate that used a modified Millipore filtration technique (Millipore Corp., Bedford, Mass.). Both methods were found to be superior to the 20-ml culture method. We suggest using the Septi-Chek method as the standard protocol for the culture of dialysate as a relatively inexpensive and labor-saving recovery technique.
Evaluation of three immunofluorescence assays for culture confirmation and typing of herpes simplex virus
Authors: Lipson S M , Schutzbank T E , Szabo K
Published: January 2020

Three pairs of monoclonal antibodies, supplied in kits by Electro-Nucleonics, Inc. (ENI), The Syva Co., and Kallestad Laboratories, Inc. (KL), were evaluated for the laboratory confirmation and typing of herpes simplex virus (HSV). Of 108 coded HSV slide preparation, run in parallel with each monoclonal-antibody set, 103 were equivalent by the immunofluorescence assays. Among the five discordant isolates, three (2.8%) did not type with the KL monoclonal antibodies and two (1.9%) false-positive results occurred with the Syva typing system. All of the HSV clinical isolates tested were correctly typed with the ENI indirect immunofluorescence antigen detection system. Typing confirmation of the five discordant HSV isolates was performed by differential sensitivity to 5-bromo-2'-deoxyuridine and endonuclease cleavage analysis of the viral DNA. Use of the Syva and KL direct immunofluorescence antigen detection systems for the identification of HSV isolates is less time-consuming than use of the ENI indirect antigen detection system; however, sensitivity and specificity may be lost.
Receptor-specific agglutination tests for detection of bacteria that bind globoseries glycolipids
Authors: de Man P , Cedergren B , Enerbäck S , Larsson A C , Leffler H , Lundell A L , Nilsson B , Svanborg-Edén C
Published: January 2020

Specific binding to the globoseries of glycolipid receptors explains the adherence of uropathogenic Escherichia coli to host cells. The minimal receptor disaccharide Gal alpha 1----4Gal beta [galactose alpha (1----4)galactose beta] is recognized by most attaching clinical isolates. However, wild-type isolates can express adhesins with several different receptor specificities. Bioassays do not permit separate analysis of each receptor specificity, since the target cells contain multiple potentially receptor-active molecules. In this study, bacterial adhesins were analyzed by using receptors immobilized into latex beads in one of two ways. In one way, di- and trisaccharides were covalently linked via a spacer arm to latex beads coupled with bovine serum albumin. In the other way, receptor-active glycolipids were coated onto the bovine serum albumin-latex beads. The latex beads were subsequently used for agglutination by using type strains with known receptor specificity. The composition was optimized regarding receptor structure and size of latex beads. Gal alpha 1----4Gal beta was as active as the trisaccharide derivative Gal alpha 1----4Gal beta 1----3glucose or Gal alpha 1----4Gal beta 1----3glucosamine. Among the natural glycolipids tested, globotetraosylceramide was the most active. Subsequently, the sensitivity and specificity of the Gal alpha 1----4Gal beta-latex and globotetraosylceramide-latex reagents were compared for 733 E. coli urinary isolates. Hemagglutination of human erythrocytes was used as the positive standard. No significant difference in the specificity or sensitivity of the latex reagents was found; the sensitivity ranged from 86%, when isolates agglutinating human erythrocytes of blood groups P1 and p were included, to 93%, when those isolates agglutinating erythrocytes of blood group p were excluded. These reagents provide tools for bacterial identification in patients with urinary tract infection.
Cryptococcus neoformans var. gattii in Australia
Authors: Ellis D H
Published: January 2020

An examination of 45 clinical isolates of Cryptococcus neoformans revealed an unusually high incidence of C. neoformans var. gattii in South Australia (65%) and in the Northern Territory (95%). In assessing all the available data from Australian isolates of C. neoformans, there appeared to be an endemic focus for the incidence of C. neoformans var. gattii in the rural aboriginal population of the Northern Territory.
Comparative evaluation of commercial enzyme immunoassay kits for detection of hepatitis B seromarkers
Authors: Ratnam S , Tobin A M
Published: January 2020

The commercial hepatitis B enzyme immunoassay kits of Abbott Laboratories and Organon Teknika were compared for their sensitivity, specificity, and reproducibility in detecting the hepatitis B seromarkers hepatitis B surface and e antigens and antibodies to hepatitis B core, e, and surface antigens. With the exception of the Organon kit for antibodies to hepatitis B surface antigen, the specificity and reproducibility were about the same for both products, but the level of sensitivity was generally lower for the Organon kits; this, however, may not be critical in routine clinical application. The Organon kits have a longer shelf life and are cheaper.
Comparison of HEp-2 cell culture and Abbott respiratory syncytial virus enzyme immunoassay
Authors: Bromberg K , Tannis G , Daidone B , Clarke L , Sierra M
Published: January 2020

Respiratory syncytial virus (RSV) is the leading cause of lower respiratory tract infection in children. Rapid identification of RSV infections would allow for specific chemotherapy. We evaluated a means of rapid diagnosis, the Abbott enzyme immunoassay (EIA), by using 314 stored nasopharyngeal aspirates. RSV antigens were identified in 62 of 66 RSV culture-positive specimens. An additional 37 specimens from which RSV was not isolated were positive in the EIA. Of these, 29 were confirmed as truly positive by a blocking assay, for a total of 95 (66 + 29) positive specimens. The sensitivity of the EIA for total positive samples was 96% (91/95) versus 69% (66/95) for cell culture. The specificity of the EIA was 96% (211/219). In these stored specimens, Abbott EIA was superior to cell culture for the detection of RSV.
Spontaneous bacterial peritonitis caused by Pasteurella ureae
Authors: Noble R C , Marek B J , Overman S B
Published: January 2020

Pasteurella ureae is uncommonly isolated from patients and rarely causes disease. The case of a patient with spontaneous bacterial peritonitis caused by P. ureae is described and contrasted with other reported cases of P. ureae colonization and infection. Respiratory colonization has been described in older patients, predominantly men, with underlying lung disease consisting principally of bronchitis, bronchiectasis, and tumors. Only 11 patients (including the present patient) have been reported to have illness with P. ureae. Seven had meningitis, two had pneumonia, and one had hepatitis. Skull fracture, alcoholism, and liver disease are common antecedents of P. ureae infection.
Detection of metabolites by frequency-pulsed electron capture gas-liquid chromatography in serum and cerebrospinal fluid of a patient with Nocardia infection
Authors: Brooks J B , Kasin J V , Fast D M , Daneshvar M I
Published: January 2020

Serum (SR) and cerebrospinal fluid (CSF) from a patient suspected of having tuberculous meningitis were submitted to our laboratory for analysis by frequency-pulsed electron capture gas-liquid chromatography (FPEC GLC). The samples were tested for the presence of carboxylic acids, alcohols, hydroxy acids, and amines by methods described previously (C. C. Alley, J. B. Brooks, and D. S. Kellogg, Jr., J. Clin. Microbiol. 9:97-102, 1977; J. B. Brooks, C. C. Alley, and J. A. Liddle, Anal. Chem. 46:1930-1934, 1974; J. B. Brooks, D. S. Kellogg, Jr., M. E. Shepherd, and C. C. Alley, J. Clin. Microbiol. 11:45-51, 1980; J. B. Brooks, D. S. Kellogg, Jr., M. E. Shepherd, and C. C. Alley, J. Clin. Microbiol. 11:52-58, 1980). The results were different from previous FPEC GLC profiles of SR and CSF from patients with known tuberculous meningitis. Both the SR and CSF contained several unidentified compounds that were not previously detected in tuberculous meningitis or any of our other studies of body fluids. Nocardia brasiliensis was later isolated from the patient. Detection of these metabolites by FPEC GLC could prove to be useful for rapid diagnosis of Nocardia disease, and their identification will provide a better understanding of metabolites produced by Nocardia sp. in vivo.
Pseudomonas stutzeri pneumonia and septicemia in a patient with multiple myeloma
Authors: Potvliege C , Jonckheer J , Lenclud C , Hansen W
Published: January 2020

A case of septicemia caused by Pseudomonas stutzeri belonging to the unusual biotype Vb-3 in a patient with multiple myeloma is described. The origin of the septicemia was attributed to a community-acquired pneumonia. The bacteriology and pathogenicity of P. stutzeri are reviewed.
Production and characterization of yeast killer toxin monoclonal antibodies
Authors: Polonelli L , Morace G
Published: January 2020

Monoclonal antibodies were obtained after fusion of mouse myeloma cells with spleen cells isolated from mice primed with a crude extract of yeast killer toxin produced by a strain of Hansenula anomala. Hybridomas were selected by specific immunoassay reaction of their fluid with crude yeast killer toxin extract. Among the monoclonal antibodies, which were characterized by the Western blot technique, one (designated KT4) proved to have precipitating properties, thus permitting the neutralization of the killer activity of the toxin. Experiments in double immunodiffusion showed that monoclonal antibody KT4 produced homologous precipitin bands by reacting with either the crude toxin used as immunogen or a toxic extract of Hansenula mrakii. It is suggested that these monoclonal antibodies will be useful for the purification, characterization, and understanding of the bioactions of yeast killer toxins.
Laboratory proficiency test results on use of selective media for isolating Pseudomonas cepacia from simulated sputum specimens of patients with cystic fibrosis
Authors: Tablan O C , Carson L A , Cusick L B , Bland L A , Martone W J , Jarvis W R
Published: January 2020

Pseudomonas cepacia colonization of or infection in patients with cystic fibrosis (CF) has been associated with increased morbidity and premature death. However, current data on national incidence may be biased because of interlaboratory differences in the methods of culturing sputa of patients with CF. We conducted three tests to evaluate the proficiency of microbiology laboratories at CF centers in identifying and isolating P. cepacia and to assess the value of using selective media for P. cepacia (P. cepacia agar and oxidation-fermentation polymyxin-bacitracin-lactose medium [OFPBL]) to recover P. cepacia from specimens simulating sputa of patients with CF. In test 1, we evaluated the proficiency of laboratories in identifying P. cepacia. Of 111 laboratories tested, 105 (95%) correctly identified P. cepacia. In test 2, we evaluated the proficiency of laboratories in isolating P. cepacia from simulated CF sputum specimens. Only 36 (32%) of 115 laboratories detected P. cepacia. Recovery of the microorganism was associated with the use of P cepacia agar or OFPBL; 14 (95%) of 15 laboratories using P. cepacia agar or OFPBL (or both) versus 22 (22%) of 100 laboratories not using either medium recovered P. cepacia (P less than 0.0001, Fisher exact test, one tailed). Laboratories failing test 2 were requested to use a selective medium for P. cepacia in a repeat test; 73 (97%) of 75 laboratories using P. cepacia agar or OFPBL (or both) versus 0 of 4 laboratories not using either medium detected P. cepacia (P less than 0.0001, Fisher exact test, one tailed). Our studies show that (i) microbiology laboratories at CF centers are proficient in identifying P. cepacia, and (ii) the use of selective media for P. cepacia enhances recovery of the microorganism in simulated sputum specimens. Therefore, we recommend the use of selective media for P. cepacia in laboratories processing sputa of patients with CF.
Gram-negative bacilli as nontransient flora on the hands of hospital personnel
Authors: Guenthner S H , Hendley J O , Wenzel R P
Published: January 2020

The possibility that gram-negative bacilli (GNB) are part of the nontransient flora on hands was examined by using a broth rinse technique to detect low titers of GNB after a hygienic hand wash with soap and water. A total of 100 nurses who had direct patient contact and 40 controls without patient contact had a similar rate of recovery of GNB (46 and 55%, respectively). GNB persisted on the hands of 10 nurses throughout five successive hand washes with soap and water. Hand cultures were obtained daily from 12 nurses before and after a work shift in a surgical intensive care unit. GNB were recovered from 57% of individuals before patient contact and from only 24% after the work shift. Nontransient GNB on the hands of hospital personnel are a potential reservoir for hospital strains, and patient contact is not an obvious source for the acquisition of nontransient GNB.
Bacteriologic and clinical study of Bacteroides oris and Bacteroides buccae
Authors: Johnston B L , Edelstein M A , Holloway E Y , Finegold S M
Published: January 2020

We characterized clinical isolates previously identified in our laboratory as Bacteroides ruminicola, the human strains of which are now classified as Bacteroides oris and Bacteroides buccae. A total of 72 isolates (55 B. buccae isolates and 17 B. oris isolates) recovered over a 10-year period were studied. They were differentiated from each other by special-potency antibiotic disks and the RapID-ANA system. The two organisms were associated with a variety of infections, the majority being pleuropulmonary (29.2%) and infections of the head and neck region (27.8%). The infections were always polymicrobial, usually with more than five organisms per specimen. A total of 44% of the B. oris strains and 27% of the B. buccae strains were resistant to penicillin G (breakpoint, 2 U/ml), and this correlated with the presence of beta-lactamase. Although B. oris and B. buccae are found with some frequency in human infections, they are present primarily as components of a mixed flora.
Isolation of Simonsiella sp. from a neonate
Authors: Whitehouse R L , Jackson H , Jackson M C , Ramji M M
Published: January 2020

A member of the genus Simonsiella, presumptively identified as S. muelleri, was isolated from a gastric aspirate taken from a neonate 15 min postpartum. The neonate showed a dental cyst and early eruption of teeth, confirmed by mandibular X ray. The morphological features, cultural characteristics, and antimicrobial susceptibility of the isolate are presented.
Production of Vi monoclonal antibodies and their application as diagnostic reagents
Authors: Tsang R S , Chau P Y
Published: January 2020

Serum antibodies to Vi antigen were detected in mice immunized with the purified antigen but not with Vi-bearing Salmonella typhi whole cells. Fusion of the spleen cells from one of the Vi antibody-producing mice with NSI myeloma cells produced four stable hybridomas that secreted antibodies to Vi. Monoclonal antibodies from these four clones were all of the immunoglobulin G class and, as determined by competition, appeared to have the same epitope specificity. Despite their immunoglobulin G nature, mouse ascitic fluids induced by one of the hybridomas strongly agglutinated the Vi-positive strains of S. typhi, S. dublin, and Citrobacter strain 5396/38. Thus, 10 clinical isolates of S. typhi but not 98 strains of other bacteria were reactive in slide agglutination tests with the monoclonal antibodies.
Antibodies to various bacterial cell wall peptidoglycans in human and rabbit sera
Authors: Wergeland H I , Endresen C
Published: January 2020

Sera from patients with verified systemic staphylococcal infection contained antibodies reactive with peptidoglycan (PG) from Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Micrococcus lysodeikticus, Bacillus subtilis, and Escherichia coli. The presence of anti-PG cross-reactive antibodies was verified in patient sera by inhibition studies with the various bacterial PGs. Antibodies to nonstaphylococcal PGs were also elevated in sera from rabbits immunized with S. aureus PG. Antibodies to S. aureus PG were removed with the synthetic peptide analogs of S. aureus PG, the L-Lys-D-Ala-D-Ala, L-Lys-D-Ala, and (Gly)5 determinants, as well as with an S. aureus PG peptide fragment containing the determinants D-Ala-D-Ala and L-Lys-D-Ala. Isolated antibodies to the PG peptides, both synthetic and native, were reactive with S. aureus and S. epidermidis PGs. The antibodies to the D-Ala-D-Ala and the L-Lys-D-Ala determinants were also reactive with S. pyogenes PG, but not with PGs from M. lysodeikticus, B. subtilis, and E. coli.
Cellular and humoral immune response to a phenolic glycolipid antigen (PhenGL-I) in patients with leprosy
Authors: Koster F T , Scollard D M , Umland E T , Fishbein D B , Hanly W C , Brennan P J , Nelson K E
Published: January 2020

The ability of phenolic glycolipid I (PhenGL-I) of Mycobacterium leprae to stimulate in vitro lymphocyte proliferation (LP) was tested in cultures of peripheral blood cells from 42 patients with leprosy in Chicago and Thailand, 9 individuals with household contact in Thailand, and 10 unexposed North American controls. Only 10 responders (24%) were found among the patients, and the degree of LP was small. Responders were found among patients with lepromatous (18%) or tuberculoid (30%) leprosy without relation to age, complications, duration of treatment, or lepromin responsiveness. The specificity of the response was supported by a lack of response to two other glycolipids, by responses by T cells but not B cells, and by the observation that three of four responders tested maintained their responses to PhenGL-I for at least 1 year. Serum immunoglobulin M (IgM) and IgG antibodies were measured in the same patients by using PhenGL-I or its terminal monosaccharide conjugated to a bovine serum albumin carrier in an enzyme-linked immunosorbent assay. The presence of IgM antibody correlated negatively with LP to lepromin and to PhenGL-I in patients with tuberculoid leprosy. We conclude that circulating T cells from some leprosy patients proliferate in the presence of PhenGL-I in vitro, but the response is weak, possibly due to concomitant suppression or inhibition. The predominance of IgM antibody to PhenGL-I may be related to a lack of a T-helper-cell-mediated switch to IgG antibody response.
Legionella pneumophila serogroup 12 isolated from human and environmental sources
Authors: Thacker W L , Wilkinson H W , Benson R F , Brenner D J
Published: January 2020

A Legionella-like organism (strain 570-CO-H [= ATCC 43290]) isolated from the lung tissue of a patient with pneumonia was shown by growth, as well as physiological, serological, and genetic characteristics, to belong to a new Legionella pneumophila serogroup, serogroup 12. Two additional strains were detected with antiserum specific for strain 570-CO-H. These strains were isolated from environmental sources.
Detection of antibody to murine cytomegalovirus by enzyme-linked immunosorbent and indirect immunofluorescence assays
Authors: Classen D C , Morningstar J M , Shanley J D
Published: January 2020

We have compared murine cytomegalovirus (MCMV) antibody determination by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence assay. A comparison of antibody detection with 146 serum samples at a 1:20 dilution showed 100% agreement (60 negatives and 86 positives) between the assays. There was close agreement of endpoint determinations of sera by both methods. After experimental MCMV infection, antibody to MCMV was detected by both assays as early as day 7, and high titers persisted as late as 6 months. In contrast to immunocompetent littermates, athymic nude mice did not develop antibody after infection. Mice lacking antibody detectable by ELISA were susceptible to lethal MCMV challenge. In a survey of animals from five commercial sources, MCMV antibody was not detected unless mice were experimentally infected. MCMV antibody determination by ELISA is a convenient method, comparable to the indirect immunofluorescence assay in sensitivity and specificity.
Serogroup F, a new capsule serogroup of Pasteurella multocida
Authors: Rimler R B , Rhoades K R
Published: January 2020

Four capsule serogroups (A, B, D, and E) have been described by using passive hemagglutination tests. Serogroups A and D predominant in pasteurelloses of avian species. A new capsule serogroup of Pasteurella multocida has been isolated from turkeys in Arkansas, California, Indiana, Iowa, Missouri, New Jersey, and Virginia. Strains belonging to the new serogroup were somatic serotype 1, 3, 7, or 12, and they varied in virulence for mice and poults. Antisera made in rabbits passively protected mice against challenge with the same serogroup regardless of somatic serotype.
Evaluation of the Bio-EnzaBead test for syphilis
Authors: Moyer N P , Hudson J D , Hausler W J
Published: January 2020

The sensitivity and specificity of the Bio-EnzaBead test for syphilis and the fluorescent treponemal antibody-absorption (FTA-ABS) test were determined by examining 262 serum samples, including 202 serum samples from patients with confirmed syphilis in various stages. Overall correlation with patient history was 95.8% with both tests. False-negative Bio-EnzaBead tests occurred in 9 of 86 (10.5%) cases of late-latent syphilis (greater than 2 years) and in 1 of 38 (2.6%) cases for which the stage of disease could not be determined. False-negative FTA-ABS tests occurred in 5 of 86 (5.8%) cases of late-latent syphilis (greater than 2 years) and in 2 of 38 (5.3%) cases for which the stage of disease could not be determined. One false-positive test occurred with Bio-EnzaBead, and the cause could not be determined. The reproducibility of the Bio-EnzaBead test was excellent when spectrophotometric readings were calibrated against either air or substrate blanks. The Bio-EnzaBead test for syphilis is a suitable alternative to the FTA-ABS test.
Detection of Breda virus antigen and antibody in humans and animals by enzyme immunoassay
Authors: Brown D W , Beards G M , Flewett T H
Published: January 2020

Enzyme immunoassays were developed for the detection of Breda virus antibody and antigen. Cattle sera collected in the United Kingdom were found to have a high prevalence of antibody (55%) to Breda virus when examined in a competitive enzyme-linked immunosorbent assay. A low prevalence of antibody was found in pigs (2.2%), and no antibody was found in sheep or goat sera. No antibody to either Breda virus or Berne virus was detected in human sera collected from veterinarians and farm workers. Only 1 of 430 human fecal specimens (0.2%) contained Breda virus antigen detectable by enzyme-linked immunosorbent assay.
Immunospecific T-lymphocyte stimulation by membrane proteins from Francisella tularensis
Authors: Sandström G , Tärnvik A , Wolf-Watz H
Published: January 2020

Membranes from a capsule-deficient mutant of the live-vaccine strain of Francisella tularensis (F. tularensis LVS) were treated with N-lauroyl sarcosinate (Sarkosyl; CIBA-GEIGY Corp., Summit, N.J.). When the Sarkosyl-insoluble fraction was heated in the presence of sodium dodecyl sulfate and mercaptoethanol and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, several polypeptides were distinguished. Four major polypeptides were eluted from the gel, each of which stimulated lymphocytes from tularemia-vaccinated individuals but not from nonvaccinated individuals. The stimulation occurred mainly in T lymphocytes. Radioactive labeling of surface proteins of the capsule-deficient bacteria indicated that at least two of the four polypeptides originated from outer membrane proteins. The results suggest that several membrane proteins of F. tularensis LVS induce a specific T-lymphocyte response.
Clinical features and therapeutic interventions in 17 cases of Bacillus bacteremia in an immunosuppressed patient population
Authors: Cotton D J , Gill V J , Marshall D J , Gress J , Thaler M , Pizzo P A
Published: January 2020

We retrospectively examined episodes of Bacillus bacteremia at a hospital with a large proportion of immunosuppressed patients. Seventeen episodes in 9.5 years met our case definition: two of two bottles of one blood culture or one of two bottles of two or more separately obtained blood cultures drawn on the same date. During the same period, there were 59 additional episodes in which a single blood culture had only one of two bottles positive for Bacillus species. Only 2 of 59 such episodes resulted in recurrent bacteremia (3%), as compared with 5 of 17 episodes meeting our case definition (29%) (P = 0.004). In four of five episodes complicated by recurrent bacteremia and in which appropriate antibiotics were used, a Hickman-Broviac catheter was in place and was not removed. We suggest that our case definition permits the differentiation of infection from contamination based on outcome and that patients with Bacillus bacteremia have chronic venous catheters removed as well as receive antibiotic treatment.
Atypical isolates of Brucella abortus from Canada and the United States characterized as dye sensitive with M antigen dominant
Authors: Ewalt D R , Forbes L B
Published: January 2020

A total of 41 Brucella isolates, examined by standard biotyping procedures, were found to be similar to Brucella abortus biovar 2 in dye sensitivity but had a dominant M antigen. Oxidative metabolic tests performed on 39 of the isolates confirmed them as B. abortus. Additional biochemical and bacteriophage susceptibility studies were performed on 35 of the isolates. The isolates had identical reactions in the various tests, except for one isolate which was resistant to lysis by all phage strains used. Two isolates were injected into guinea pigs and shown to be virulent. The isolates described in this study appear similar to atypical Brucella isolates previously reported in the United Kingdom and the United States and may form the basis of a new biovar, B. abortus biovar 10.
DNA probes for detection and identification of Mycoplasma pneumoniae and Mycoplasma genitalium
Authors: Hyman H C , Yogev D , Razin S
Published: January 2020

DNA probes specific for Mycoplasma pneumoniae and Mycoplasma genitalium were selected from genomic libraries prepared in pUC13. The 32P-labeled probes could detect, by dot blot hybridization, down to about 0.1 ng of the specific mycoplasma DNA or 10(5) CFU. Biotinylation of probe decreased the sensitivity of detection and produced nonspecific background reactions with nonhomologous DNAs. Sulfonation of probe yielded a similar level of sensitivity with less background.
Rapid screening assay for phospholipase C activity in mycoplasmas
Authors: De Silva N S , Quinn P A
Published: January 2020

A screening assay for phospholipase C using a chromogenic substrate incorporated into agar medium is described. The assay directly visualizes phospholipase C activity of mycoplasma lysates and membranes on agar plates, or the activity may be measured by spectrophotometry. The results from the assay confirm the presence in Ureaplasma urealyticum of phospholipase C, which is predominantly localized in the membrane fraction. The procedure has the potential to screen phospholipase C activity in other mycoplasmas and microorganisms in general.
Effect of source of Mueller-Hinton agar on detection of oxacillin resistance in Staphylococcus aureus using a screening methodology
Authors: Hindler J A , Warner N L
Published: January 2020

Dehydrated Mueller-Hinton agar from five different manufacturers was used to prepare plates containing 4% NaCl and 6 micrograms of oxacillin per ml to screen for oxacillin resistance in Staphylococcus aureus. A total of 137 isolates which included 109 oxacillin-resistant isolates obtained from at least eight geographic areas were examined. Results were compared to MICs obtained by using a standard broth microdilution method. Results obtained with screening plates prepared with dehydrated media from three of the manufacturers showed 100% correlation with MICs in detecting oxacillin-resistant S. aureus, and plates prepared with the remaining two media identified oxacillin resistance in 90 and 99% of the resistant isolates, respectively. None of the oxacillin-susceptible isolates grew on any of the screening plates. This study demonstrated that oxacillin screening plates are reliable for detecting oxacillin-resistant S. aureus; however, the source of Mueller-Hinton agar can influence the results.
Selected characteristics of pathogenic and nonpathogenic strains of Bacteroides gingivalis
Authors: Grenier D , Mayrand D
Published: January 2020

Strains of Bacteroides gingivalis were compared for the presence of properties associated with pathogenicity. Some strains were infectious in pure culture in an in vivo model (guinea pig), and all but one of these were more collagenolytic than those which failed to cause lesions in guinea pigs. However, other factors seem to be necessary for the induction of an infection in this animal model.
Comparison of three rapid diagnostic techniques for detection of respiratory syncytial virus from nasal wash specimens
Authors: Chonmaitree T , Bessette-Henderson B J , Hepler R E , Lucia H L
Published: January 2020

We report results of three rapid tests for respiratory syncytial virus antigen detection. An immunofluorescence assay using commercial antibody and two commercial enzyme immunoassays (Ortho Diagnostics, Inc., Raritan, N.J., and Abbott Laboratories, North Chicago, Ill.) were applied to 199 nasal wash specimens. The Abbott enzyme immunoassay was the most sensitive technique, with a sensitivity of 93.8%. The specificities of the three techniques were comparable and greater than 95%. The availability of reliable rapid diagnostic techniques will allow for better care of infants with severe respiratory syncytial virus infection.
Differential immunoglobulin G subclass antibody titers to respiratory syncytial virus F and G glycoproteins in adults
Authors: Wagner D K , Nelson D L , Walsh E E , Reimer C B , Henderson F W , Murphy B R
Published: January 2020

Two respiratory syncytial virus glycoproteins, F and G, which differ substantially in the amount of glycosylation were used as antigens in an enzyme-linked immunosorbent assay to determine immunoglobulin G (IgG) subclass titers in 30 experimentally infected healthy adults. The titers of antibodies to the F glycoprotein achieved in postinfection sera were highest in the IgG1 subclass, whereas those to the G glycoprotein were highest and comparable in the IgG1 and IgG2 subclasses. The high IgG2 response to the G glycoprotein suggests that it is seen by the immune system as a polysaccharide antigen, a hypothesis consistent with its large carbohydrate content.
Clinical significance of virulence-related assay of Yersinia species
Authors: Noble M A , Barteluk R L , Freeman H J , Subramaniam R , Hudson J B
Published: January 2020

During the 42-month period from June 1982 through December 1985, 215 fecal specimens from 171 patients were found to be positive for yersiniae by using a combination of CIN agar and cold enrichment. Isolates were tested for markers of virulence including carriage of a plasmid 42 megadaltons in size, calcium dependence, autoagglutination, Congo red uptake, pyrazinamidase activity, fermentation of salicin, and hydrolysis of esculin. The results were correlated to symptoms in patients. A total of 80 Yersinia enterocolitica and 52 Y. enterocolitica-like strains (42 Y. frederiksenii, 8 Y. intermedia, and 2 Y. kristensenii) were examined. Positive virulence-related tests were as follows (for Y. enterocolitica, Y. frederiksenii, Y. intermedia, and Y. kristensenii, respectively): pyrazinamidase negativity, 12.5, 0, 0, and 50%; Congo red positivity, 5, 7.1, 87.5 and 0%; calcium dependence, 3.8, 0, 0, and 0%; autoagglutination positivity, 8.8, 0, 0, and 0%; carriage of the 42-megadalton plasmid, 28.6, 73.2, 5.7, and 0; salicin and esculin negativity, 12.5, 0, 0, and 50%. The isolates recovered from symptomatic patients were characterized in relation to the presenting symptoms. Isolates from 12 of 32 (37.5%) patients with acute-onset diarrhea and 9 of 30 (30.0%) patients with chronic symptoms expressed at least one virulence feature. No individual test or group of tests was consistently associated with onset or either type of symptoms. Routine testing of plasmid carriage, uptake of Congo red, calcium dependence, autoagglutination, and pyrazinamidase activity did not appear to provide information that would link the presence of symptoms with the virulence potential of fecal isolates of yersiniae.
Comparison of fluorescent-antibody-to-membrane-antigen test, indirect immunofluorescence assay, and a commercial enzyme-linked immunosorbent assay for determination of antibody to varicella-zoster virus
Authors: Landry M L , Cohen S D , Mayo D R , Fong C K , Andiman W A
Published: January 2020

The demand for sensitive and specific assays to determine immune status to varicella can be expected to increase with the anticipated availability of a varicella-zoster virus vaccine for use in nonimmune adults, especially health care personnel, and in immunosuppressed children. Although the fluorescent-antibody-to-membrane-antigen (FAMA) test remains the reference standard to which other tests are compared, simpler alternative assays are needed. In this study, the FAMA was compared with a simple indirect immunofluorescence assay (IFA) and a commercially available enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to varicella-zoster virus. One hundred and twelve serum samples were screened by the FAMA test and IFA at a 1:5 dilution, and 100% agreement was found. Of these samples, 101 were available for testing by ELISA, and identical results were obtained with 97 samples (96% agreement). When the samples were screened at a 1:2 dilution, 99 of 101 results agreed. In addition, 31 spinal fluid samples were tested by all three methods. When screening was at a 1:2 dilution, there was 96.8% agreement between the FAMA test and IFA. When the cutoff value established for sera was used for the spinal fluid samples, there was 90.3% agreement between the ELISA and the FAMA test. Thus, both IFA and ELISA can be considered sensitive and specific alternatives to the FAMA test, and in addition, both use commercially available reagents.
Impact of prolonged incubation on disk diffusion susceptibility test results for Staphylococcus aureus
Authors: Mulligan M E , Citron D M , Kwok R Y , Wheelock J P , Farrohi F K , Hindler J A , Johnston L
Published: January 2020

Because strains of Staphylococcus aureus that are resistant to penicillinase-resistant penicillins may be difficult to detect in the clinical laboratory, a variety of changes in methodology have been suggested to increase their detection. In 1984, the West Los Angeles Veterans Administration Medical Center experienced an increase in clinically significant strains of oxacillin-resistant S. aureus. To insure that such strains would not be missed by the disk diffusion test methods employed for routine testing, changes in methodology were insituted. These included interpreting zone diameters around oxacillin disks at 48 h of incubation. We collected 139 isolates from patients thought to have oxacillin-resistant S. aureus based on these test results and later retested the isolates using microdilution MIC testing. Only 85 isolates (61%) had microdilution oxacillin MICs of greater than or equal to 8.0 micrograms/ml, whereas 54 (39%) had oxacillin MICs of less than or equal to 2.0 micrograms/ml. A review of medical records revealed that in 1 year there were 98 patients with isolates appearing resistant by disk diffusion but not confirmed by microdilution MICs; many patients were placed in isolation and treated with specific antimicrobial agents. We conclude that incubation of oxacillin disk diffusion tests for longer than 24 h in conjunction with disregard for resistance to other classes of antimicrobial agents may result in an unacceptably high degree of false resistance results. Because the resistance of S. aureus has important therapeutic and infection control implications, it is necessary to recognize problems that may result in ambiguous or inaccurate susceptibility results.
Monoclonal antibodies to gp110 and gp41 of human immunodeficiency virus
Authors: Gosting L H , McClure J , Dickinson E S , Watanabe S M , Shriver K , Goldstein L C
Published: January 2020

Six mouse hybridomas secreting monoclonal antibodies specific for the glycoproteins of human immunodeficiency virus were developed. All six antibodies reacted by radioimmunoprecipitation with the glycoprotein precursor of 150,000 daltons as well as one of the proteolytic processing products of 110,000 daltons (gp110) or 41,000 daltons (gp41). Recombinant polypeptides spanning the env coding region were used to locate epitopes on the glycoprotein molecule. The panel of antibodies detected two distinct epitopes of gp41 and one epitope of gp110. We used the antibodies in indirect immunofluorescence assays to evaluate 13 clinical isolates of human immunodeficiency virus from diverse geographic regions, and we found that the gp110 epitope was recognized on all tested isolates, whereas the two gp41 epitopes were detected on 10 of 13 and 4 of 13 isolates.
Media for study of growth kinetics and envelope properties of iron-deprived bacteria
Authors: Kadurugamuwa J L , Anwar H , Brown M R , Shand G H , Ward K H
Published: January 2020

Ion-exchange chromatography was used to remove iron from complex and chemically defined laboratory media. The kinetics of metal cation removal from the media was investigated by using atomic absorption spectrophotometry, and the results indicated that over 90% of the iron could be eliminated from certain complex media by this treatment. The treated medium was used for growth studies in a gram-positive and a number of gram-negative organisms that were isolated from infections in humans. High-molecular-weight outer membrane proteins that are known to be induced under iron-depleted growth conditions (iron-regulated membrane proteins) were observed when a number of gram-negative pathogens were cultivated in the treated media. Iron uptake by Staphylococcus aureus varied, depending on the iron content of the medium.