bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of an enzyme-linked immunosorbent assay with indirect hemagglutination and hemagglutination inhibition for determination of rubella virus antibody: evaluation of immune status with commercial reagents in a clinical laboratory
Authors: Truant A L , Barksdale B L , Huber T W , Elliott L B
Published: January 2020

Comparative evaluations of immune status for rubella virus are described for enzyme-linked immunosorbent assay, hemagglutination inhibition, and indirect hemagglutination. A 92.1% agreement between enzyme-linked immunosorbent assay and indirect hemagglutination assay was demonstrated for rubella immune status. Enzyme-linked immunosorbent assay and hemagglutination inhibition demonstrated a 92.6% agreement and were compared in an attempt to define the quantitative usefulness of comparisons of single sera for determining immune status. These data support the relative lack of correlation between single enzyme-linked immunosorbent assay and hemagglutination inhibition quantitative values. Enzyme immunoassay was, however, an acceptable alternative to hemagglutination inhibition for the determination of immune status to rubella virus.
Quantities of Clostridium botulinum organisms and toxin in feces and presence of Clostridium botulinum toxin in the serum of an infant with botulism
Authors: Paton J C , Lawrence A J , Steven I M
Published: January 2020

A 7-week-old boy presented with symptoms and signs characteristic of infant botulism, and the diagnosis was confirmed by the detection of Clostridium botulinum type A organisms and toxin in the feces. The levels of organisms and toxin in the feces were measured throughout the 81-day period in hospital. The maximum levels detected were 2.46 x 10(8) C. botulinum type A colony-forming units and 64,000 mouse 100% lethal doses of type A toxin per g (wet weight) of feces. C. botulinum toxin was also detected in two samples of the patient's serum, collected 3 and 10 days after admission. Improvement in the patient's clinical condition occurred before the levels of organisms and toxin in the feces reached their maxima. A slight improvement may also have occurred while toxin was still present in the serum.
Bacteremic infections caused by nontypable Haemophilus influenzae in patients with dysgammaglobulinemia
Authors: Musher D M , Wallace R J
Published: January 2020

Two patients with dysgammaglobulinemia had bacteremic infections due to nontypable Haemophilus influenzae. The regular use of counterimmunoelectrophoresis or the more careful use of existing agglutination techniques to serotype Haemophilus isolates may identify nontypable H. influenzae in this clinical setting more frequently than has been though possible.
Coinfections of Mycoplasma pneumoniae and Legionella pneumophila with influenza A virus
Authors: Renner E D , Helms C M , Johnson W , Tseng C H
Published: January 2020

Serum samples from patients with documented influenza A virus infections were examined for antibodies to Legionella pneumophila and Mycoplasma pneumoniae to determine whether simultaneous or sequential infections with L. pneumophila and M. pneumoniae were complicating factors in influenza. When the frequency of copositivity of sera to influenza A virus and L. pneumophila was compared with the expected frequency for each infection alone, the difference was not statistically significant. However, when the frequency of copositivity of sera to influenza A virus and M. pneumoniae was compared with the expected frequency for each infection alone, there was a statistically significant (P less than 0.005) absence of coincident titers. Seasonal variations and differences in relative age frequencies for the two infections may partially explain the absence of coinfections. These data also suggest that in patients with either M. pneumoniae or influenza A virus infection, some type of protective mechanism which prevents coinfections with these organisms is present.
Microbiology of a major foodborne outbreak of gastroenteritis caused by Yersinia enterocolitica serogroup O:8
Authors: Shayegani M , Morse D , DeForge I , Root T , Parsons L M , Maupin P S
Published: January 2020

Gastrointestinal disorders of varying severity were observed in 239 (53%) of 455 campers and staff members at a coed summer camp in Sullivan County, New York, during July 1981. Five of seven hospitalized patients had appendectomies before the disease was recognized as yersiniosis. Yersinia enterocolitica serogroup O:8 (American strain) was isolated from 37 (54%) of 69 persons examined, including the head cook and 3 others of the 11-person kitchen staff. Of 48 food, water, and environmental samples collected from the camp area, Y. enterocolitica isolates belonging to the same serogroup and biogroup as the human isolates were recovered from dissolved powdered milk, a milk dispenser, and turkey chow mein. This laboratory finding supported the epidemiological data indicating a correlation between consumption of these foods and illness. Y. enterocolitica isolates of the same biogroup as the O:8 isolates but belonging to serogroup O:34 were also isolated from six campers and two samples of dissolved powdered milk. Pathogenicity studies on the Yersinia isolates were performed with three in vitro tests (calcium dependency, autoagglutination, and HeLa cell infection) and one in vivo test (intraperitoneal challenge of mice). Most of the serogroup O:8 human isolates and the chow mein isolate were positive in all four tests. Milk isolates of serogroup O:8 were positive in the in vitro tests but were relatively avirulent in mice, whereas serogroup O:34 isolates, regardless of source, were negative in all four tests.
Automatic and manual latex agglutination tests for measurement of cholera toxin and heat-labile enterotoxin of Escherichia coli
Authors: Ito T , Kuwahara S , Yokota T
Published: January 2020

Automated and manual latex agglutination methods were employed to measure cholera toxin (CT), heat-labile enterotoxin (LT) of Escherichia coli, and their subunits A and B. Dow polystyrene latex particles (diameter, 0.22 microns) and polystyrene-chlorostyrene latex particles (diameter, 1 micron) were sensitized by rabbit-specific immunoglobulin for each antigen and used as the reagents of the automated and manual agglutination tests, respectively. Automated agglutination was performed by a nephelometric assay system measuring time-dependent differences of light scattering due to agglutination, and manual latex agglutination was carried out in microtiter plates. As low as 1,000 and 31 pg of CT per ml were estimated by the automated and manual agglutination tests, respectively. Using these methods, the amount of CT and LT was measured in several clinical isolates of Vibrio cholerae and E. coli. Furthermore, it was discovered that cyclic AMP is not essential for the production of CT by measuring the amount of the toxin in numbers of cyclic AMP-dependent mutants of V. cholerae (with the agglutination tests).
Typing of clinical herpes simplex virus isolates with mouse monoclonal antibodies to herpes simplex virus types 1 and 2: comparison with type-specific rabbit antisera and restriction endonuclease analysis of viral DNA
Authors: Peterson E , Schmidt O W , Goldstein L C , Nowinski R C , Corey L
Published: January 2020

A total of 122 clinical isolates of herpes simplex virus (HSV) from 107 patients were typed by using an indirect immunoperoxidase technique with commercially available type-specific rabbit antisera, recently developed mouse monoclonal antibodies to HSV types 1 and 2, and restriction endonuclease analysis of viral DNA. With the commercially available type-specific rabbit antisera, 34% of clinical HSV isolates were of indeterminate type; 63% of them were typed as HSV type 1 and 37% as HSV type 2 by using monoclonal antibody and restriction enzyme typing systems. Typing by immunofluorescence assay with the monoclonal antibodies gave identical results to those obtained by restriction enzyme analysis. Simultaneous infection with both HSV types was demonstrated by monoclonal antibody typing in five isolates from three patients. These findings were subsequently confirmed by plaque purification and restriction endonuclease analysis of viral DNA. Monoclonal antibodies were as sensitive as restriction enzyme analysis for the typing of clinical HSV isolates. Because of their simplicity, they are more amenable to use in clinical laboratories than is restriction endonuclease analysis.
Rapid identification of Staphylococcus aureus and Streptococcus pneumoniae from blood cultures
Authors: Knight R G , Shlaes D M
Published: January 2020

Simultaneous application of the lysostaphin sensitivity test for identification of Staphylococcus aureus and the deoxycholate test for the identification of Streptococcus pneumoniae was evaluated for reliability in rapid identification (1 h) of these organisms from blood cultures by using BACTEC 6B and 7C bottles. The procedure was applied to 127 cultures, 74 lysostaphin tests and 53 deoxycholate tests. Lysostaphin-tested organisms included 23 S. aureus, 40 Staphylococcus epidermidis, 1 Listeria sp., 1 Peptococcus sp., 2 Micrococcus sp., 2 diphtheroids, and 5 mixed cultures. Deoxycholate-tested organisms included 14 S. pneumoniae, 32 Streptococcus spp. (not S. pneumoniae), 1 Listeria sp., 1 Peptococcus sp., 2 diphtheroids, and 3 mixed cultures. Analysis of the data revealed positive and negative predictive values of 95.8 and 94%, respectively, for the lysostaphin test and 100 and 97.4%, respectively, for the deoxycholate test. The combined tests provide rapid reliable identification or elimination of S. aureus and S. pneumoniae in blood cultures.
Microtiter solid-phase radioimmunoassay for detection of human calicivirus in stools
Authors: Nakata S , Chiba S , Terashima H , Sakuma Y , Kogasaka R , Nakao T
Published: January 2020

A microtiter solid-phase radioimmunoassay (RIA) was developed for detection of human calicivirus in stool specimens. Seventy-eight stool specimens were tested by RIA. All 17 specimens positive for human calicivirus by electron microscopy (EM) were also positive by RIA. In addition, of 21 specimens obtained from an outbreak of caliciviral gastroenteritis, 11 were positive by RIA but negative by EM. Of 20 specimens positive for rotavirus by EM and 20 nondiarrheic specimens with no virus, 2 and 1, respectively, were positive by RIA but were subsequently shown to be falsely positive by a blocking test. There was no cross-reaction between human and feline caliciviruses. Thus, the test was more sensitive than EM and, with an appropriate blocking test, was specific for human calicivirus. It might be especially useful for screening large numbers of stool specimens.
Evaluation of a computer-assisted, kinetics-based enzyme-linked immunosorbent assay for detection of coronavirus antibodies in cats
Authors: Barlough J E , Jacobson R H , Downing D R , Marcella K L , Lynch T J , Scott F W
Published: January 2020

A computer-assisted, kinetics-based enzyme-linked immunosorbent assay was adapted for the detection of coronavirus antibodies in feline serum. An alkaline antigen diluent (carbonate-bicarbonate buffer, pH 9.6) used in initial experiments produced diffuse, nonspecific color reactions in both viral and control antigen cuvettes which were correlated, paradoxically, with coronavirus antibody levels in test sera. These interfering reactions were minimized by use of lower-pH antigen diluents such as water and phosphate-buffered saline. Background kinetics-based enzyme-linked immunosorbent assay reactivity directed against a noncoronaviral component of antigen tissue culture fluids could then detected in numerous sera, particularly in samples with lower titers. Much of this reactivity was shown to be associated with bovine gamma globulins in cell culture fluid. It was not serum lot or species specific, since a variety of bovine serum lots as well as individual lots of serum from other mammalian and avian species reacted. Reactivity was markedly reduced when cells for antigen preparation were grown in gamma globulin-free bovine serum. Generation of corrected slope values from the kinetics-based enzyme-linked immunosorbent assay made it possible to correct for residual background reactivity in individual test sera and thus eliminate a potentially major source of false-positive reactions. Collectively, these studies indicated that the control of nonspecific reactivity in feline coronavirus serology is absolutely essential to obtain useful estimates of specific antibody responses.
Firefly luciferase ATP assay as a screening method for bacteriuria
Authors: Thore A , Lundin A , Anséhn S
Published: January 2020

A rapid (15 min) test for bacteriuria based on firefly luciferase analysis of bacterial ATP has been evaluated in 2,018 clinical urine specimens. The test procedure involves removal of nonbacterial ATP by treatment of urine with Triton X-100 and apyrase, extraction of bacterial ATP by boiling, and bioluminescent analysis of bacterial ATP by firefly luciferase, using a luminometer. For comparison, the widely used nitrite test was included in the study as an example of an alternative rapid chemical test. The test was set up to distinguish between specimens yielding greater than 10(5) CFU/ml and specimens yielding less than 10(5) CFU/ml. A level of 13.5 nM ATP was chosen to define the limit between negative and positive results. At this discriminatory level, 92% of specimens yielding greater than 10(5) CFU/ml and 88% of specimens yielding less than 10(5) CFU/ml were correctly classified with the luciferase method, whereas corresponding figures for the nitrite test were 55 and 99%, respectively. Of the 12% false luciferase positives, 20% were shown to contain greater than 10(5) CFU/ml on prolonged incubation, thus reducing the false-positive rate to 10%. Of the 8% false luciferase negatives, 65% had low levels of CFU in the range of 10(5) to 10(6).
Role of reduced sulfur compounds in nutrition of Propionibacterium acnes
Authors: Nielsen P A
Published: January 2020

In a study of the nutritional requirements of Propionibacterium acnes, acid hydrolyzed casein was shown to be the preferred nitrogen source in a glycerol-salts medium. Of 22 individual amino acids tested, only cysteine supported growth in the glycerol-salts medium. This indicated a requirement not only for amino nitrogen but also for reduced sulfur. In the presence of an auxiliary nitrogen source, thioglycolate or thiosulfate could replace sulfur-containing amino acids. The apparent requirement of P. acnes for reduced sulfur possibly could be satisfied in vivo by the constant availability of this substance in the form of sulfhydryl groups in the sebaceous follicle during keratinization. The relationship between P. acnes and its nutritional substrate may give this organism a selective advantage to exist in this ecological niche.
Enzyme-linked immunosorbent assay for detection of Pseudomonas aeruginosa Lipopolysaccharides
Authors: Kusama H
Published: January 2020

A double-antibody sandwich method of enzyme-linked immunosorbent assay was developed to detect lipopolysaccharides (LPS) from the eight most prevalent Pseudomonas aeruginosa serotypes (O1, O2,5,16, O3, O4, O6, O9, O10, and O11). Immunoglobulin M fractions from rabbit antisera were used as the coating antibody and as the antibody to be conjugated to an enzyme. When two fractions of LPS (I and II) obtained by Sepharose 2B column chromatography were assayed, LPS II showed 10 to 100 times more activity than LPS I; the detection level of LPS II was 0.1 ng/ml. When LPS in purified preparations or in culture filtrates was examined with both homologous and heterologous antibody systems, the same specificity pattern was demonstrated, suggesting that, in crude filtrates, antigens other than LPS do not interfere in the assay. The method described can be used to detect LPS in biological fluids.
Isolation and characterization of a seventh serogroup of Legionella pneumophila
Authors: Bibb W F , Arnow P M , Dellinger D L , Perryman S R
Published: January 2020

An environmental isolate (Chicago 8) and a clinical isolate (Dallas 5) of Legionella pneumophila were shown to have similar serological characteristics; however, these characteristics were distinct from those of L. pneumophila serogroups 1 through 6. Chicago 8, ATCC 33823, was designated as the reference strain for L. pneumophila serogroup 7. The use of Mongolian gerbils for the isolation of L. pneumophila from the environment is described. Even though guinea pigs are the animals of choice in such studies, the isolation of Chicago 8 illustrates that the use of gerbils may be a viable option when cost is a major consideration in study design.
Fusobacterium nucleatum pericarditis
Authors: Truant A L , Menge S , Milliorn K , Lairscey R , Kelly M T
Published: January 2020

A pericardial effusion was diagnosed by echocardiography in a 49 year old man who suffered acute cough, orthopnea, and chest pain. Because of a positive tuberculin skin test, mycobacteria were initially suspected as the cause of the pericarditis. The patient was therefore treated with antituberculosis drugs. The pericardial effusion failed to resolve, however, and pericardiectomy was performed. Culture of the pericardial fluid yielded pure Fusobacterium nucleatum growth. The patient responded to antibiotic therapy and was in good health 3 weeks after being discharged from the hospital. This represents the first report of F. nucleatum pericarditis.
Immune adherence hemadsorption: specific method for rapid identification of adenovirus antigens
Authors: Champsaur H , Slim A , Tournier P
Published: January 2020

An immune adherence hemadsorption test for the rapid group-specific identification of adenoviruses is described. The test was performed directly on the cell culture isolate in less than 1 h, by the sequential addition of the antiserum of any adenovirus serotype, complement, and then C3b receptor-rich human erythrocytes. Microscopic examination revealed the presence of erythrocytes adhering firmly to the infected cell sheet. The immune adherence hemadsorption test proved a specific and rapid diagnostic procedure for the detection of adenovirus group antigen in tissue cultures and was more simple than conventional tests.
Adenovirus hexon monoclonal antibody that is group specific and potentially useful as a diagnostic reagent
Authors: Cepko C L , Whetstone C A , Sharp P A
Published: January 2020

A monoclonal antibody to the adenovirus 2 hexon protein was produced and characterized as a group-specific antibody. Positive reactivity in immunoprecipitation, indirect immunofluorescence, and radioimmunoassays was observed with human, canine, swine, bovine, murine, and simian adenoviruses. This monoclonal antibody should provide a specific and sensitive diagnostic reagent for detection of all mammalian adenoviruses.
Photodynamic inactivation of pseudorabies virus with methylene blue dye, light, and electricity
Authors: Badylak J A , Scherba G , Gustafson D P
Published: January 2020

Pseudorabies virus was photoinactivated with a combination of methylene blue dye, light, and electricity. Viral suspensions were mixed with variable amounts of methylene blue dye and then were placed in a current source apparatus. Total inactivation of pseudorabies virus (1.7 X 10(6) 50% tissue culture infective doses per ml) was achieved with constant mixing, a methylene blue dye concentration of 10(-4) M, and an electrical current of 12 microA for 12 min.
Norleucine-tyrosine broth for rapid identification of Clostridium difficile by gas-liquid chromatography
Authors: Nunez-Montiel O L , Thompson F S , Dowell V R
Published: January 2020

A new medium, norleucine-tyrosine (NT) broth, was developed for rapid identification of Clostridium difficile on the basis of caproic acid and p-cresol production. The NT broth consists of 0.5% Trypticase (BBL Microbiology Systems, Cockeysville, Md.), 0.5% yeast extract (Difco Laboratories, Detroit), 0.2% L-norleucine and 0.2% L-tyrosine (wt/vol; final concentrations), and a mixture of salts. The procedure for demonstrating caproic acid and p-cresol production involves extracting NT broth cultures with ether or chloroform and analyzing the extracts with a gas-liquid chromatograph equipped with a thermal conductivity detector, as is customary for analysis of volatile fatty acids. A total of 120 strains of C. difficile from diverse geographic locations were tested by this procedure, and they all produced caproic acid and p-cresol in NT broth. No other Clostridium species or other microorganisms tested have been found to produce both products in NT broth.
Differentiation of Kingella denitrificans from Neisseria gonorrhoeae by growth on a semisolid medium and sensitivity to amylase
Authors: Odugbemi T , Arko R J
Published: January 2020

All 239 strains of Neisseria gonorrhoeae tested for sensitivity to amylase-impregnated disks showed zones of growth inhibition ranging from 15 to 30 mm in diameter. None of 52 strains of Kingella denitrificans were inhibited by the same amylase test. In addition, all N. gonorrhoeae strains grew well on a gonococcal base medium supplemented with either IsoVitaleX (BBL Microbiology Systems) or Supplement B (Difco Laboratories), whereas K. denitrificans grew adequately only on the medium with IsoVitaleX. The addition of L-cysteine or cystine to the medium with Supplement B enhanced the growth of K. denitrificans.
Absorption of Rickettsia rickettsii antibodies by Rickettsia rickettsii antigens in four diagnostic tests
Authors: Hechemy K E , Anacker R L , Carlo N L , Fox J A , Gaafar H A
Published: January 2020

Immunoabsorption profiles were determined for Rickettsia rickettsii antigens used in four confirmatory tests for detection of antibodies to Rocky Mountain spotted fever. A human serum reactive in the four tests was absorbed with each test antigen and then reexamined by all four tests. The results indicated that the whole organism and complement-fixation antigen absorbed the whole array of antibodies to R. rickettsii, whereas erythrocyte-sensitizing substance coated on latex or erythrocytes did not. With erythrocyte-sensitizing substance coated on latex there was a decrease in titer when tested by the latex, indirect hemagglutination, and complement fixation tests but none when tested by the microimmunofluorescence test. With erythrocyte-sensitizing substance coated on sheep erythrocytes there was a decrease in titer when tested by indirect hemagglutination or complement fixation but none when tested by latex or microimmunofluorescence. These findings confirm that erythrocyte-sensitizing substance contains more than a single antigenic substance and plays a major role in the complement fixation test and a minor role in the microimmunofluorescence test.
Inoculum standardization in antimicrobial susceptibility testing: evaluation of overnight agar cultures and the Rapid Inoculum Standardization System
Authors: Baker C N , Thornsberry C , Hawkinson R W
Published: January 2020

Using the Rapid Inoculum Standardization System [RISS; Minnesota Mining & Manufacturing Co. (3M Co.)], we investigated the use of overnight agar cultures in the preparation of inocula for disk diffusion and microdilution susceptibility tests and compared our results with susceptibility results obtained with the standard methods. The 3M system consists of an inoculation wand, with which a reproducible number of bacteria can be removed from an agar plate, and a diluent in which to suspend the organisms. We used 25 gram-positive cocci, 75 enteric and nonfermentive gram-negative bacilli, and 40 more fastidious bacteria (Haemophilus, Neisseria, and pneumococci) for the evaluation. The geometric mean inoculum size for all organisms tested was 9.7 X 10(7) CFU/ml by the standard method and 1.1 X 10(8) CFU/ml for the RISS. The categories of susceptibility obtained by both methods in the disk diffusion tests were comparable, as were the minimal inhibitory concentrations. We recommend that the use of overnight agar cultures and the use of the RISS to prepare inocula for susceptibility tests be acceptable alternative procedures in the standard methods for susceptibility tests.
Latex-Rickettsia rickettsii test reactivity in seropositive patients
Authors: Hechemy K E , Rubin B B
Published: January 2020

In correlating results obtained from a new latex-Rickettsia rickettsii test with results obtained from a reference microimmunofluorescence test for Rocky Mountain spotted fever, we found that for seropositive patients each microimmunofluorescence titer (up to 4,096) was usually double the median titer obtained from the latex-R. rickettsii test. The pattern of immunoserological response indicated that latex-R. rickettsii is reactive with sera containing either anti-R. rickettsii immunoglobulin M (IgM) or IgG or both. The agglutination efficiency of the new test was greater when the anti-rickettsial IgM/IgG ratio was greater than or equal to 1. Sera from late-convalescent patients were generally minimally reactive or nonreactive in the latex-R. rickettsii test unless anti-R. rickettsii IgM was present.
Comparison of four rapid methods for identification of Enterobacteriaceae from blood cultures
Authors: Malloy P J , Ducate M J , Schreckenberger P C
Published: January 2020

Positive blood cultures containing gram-negative bacilli were utilized for direct identification by two automated systems, the AutoMicrobic system (Vitek Systems, Inc., Hazelwood, Mo.) and the MS-2 (Abbott Diagnostics, Dallas, Tex.), and two commercial kits, the Micro-ID system (General Diagnostics, Warner-Lambert Co., Morris Plains, N.J.) and the same-day API 20E (Analytab Products, Plainview, N.Y.). Samples of 10 to 15 ml were aseptically removed from radiometrically positive BACTEC bottles (Johnston Laboratories, Cockeysville, Md.) and divided among four sterile tubes. The tubes were centrifuged at 107 X g for 10 min. The supernatants were centrifuged at 1,510 X g for 10 min, and pellets were tested for cytochrome oxidase by means of Pathotec strips. Oxidase-negative pellets were suspended in appropriate media as suggested by the manufacturers. All systems were inoculated, incubated, and interpreted according to the instructions of the manufacturers. The Micro-ID system was read after 4 h of incubation; the three remaining systems were read after 5 h. Results were compared with those of the 18-h API 20E inoculated from pure subcultures of the organisms. Correlation of 90% or more with the API 20E was achieved by the AutoMicrobic and Micro-ID systems. The same-day API 20E and the MS-2 demonstrated 60 and 44% correlation, respectively, with the 18-h API 20E.
Partial characterization of Yersinia intermedia bacteriocin
Authors: Sandhu K K , Bottone E J , Pisano M A
Published: January 2020

Yersinia intermedia produces a temperature-dependent (25 degrees C) bactericidal substance that is active against other Yersinia species. Crude preparations of the inhibitory substance were inactivated by chymotrypsin, trypsin, pronase, and heating but were not affected by lipolytic enzymes, chloroform, or other organic solvents. These data suggest that the active molecule is a bacteriocin of a proteinaceous nature. The molecular weight of the bacteriocin was estimated to be greater than 14,000. Exposure of agar fragments containing the active component to a pH range of 3 to 11 did not affect bactericidal activity. Bactericidal activity against the Y. frederiksenii indicator strain was shown by simultaneous and deferred antagonism and by the associative culture technique. The liquor from cell-free macerated agar fragments and broth cultures, however, were devoid of antibacterial activity.
Serogroups of Campylobacter jejuni, Campylobacter coli, and Campylobacter fetus defined by direct immunofluorescence
Authors: Hébert G A , Hollis D G , Weaver R E , Steigerwalt A G , McKinney R M , Brenner D J
Published: January 2020

Rabbits were inoculated with whole, formalinized Campylobacter jejuni, C. coli, and C. fetus cells; the C. jejuni and C. coli immunogens were identified by their DNA relatedness at the species level to the type strains of C. jejuni or C. coli. The designation C. coli was not used among the other C. jejuni strains; they were classified as hippurate-positive or hippurate-negative C. jejuni. Immunoglobulin G was isolated from the antisera and labeled with fluorescein isothiocyanate. These conjugates defined 10 serogroups of C. jejuni, 2 serogroups of C. coli, and 2 serogroups of C. fetus. Of the 316 strains of C. jejuni tested, 258 (82%) were groupable; 173 were single-serogroup strains, and 85 were multiple-serogroup strains. Of the 226 strains in C. jejuni serogroups, 223 (98.7%) were hippurate positive; of the 27 strains in C. coli serogroups, 26 (96.3%) were hippurate negative. Five strains were equally reactive in immunofluorescent staining with a conjugate for a C. jejuni serogroup and a conjugate for a C. coli serogroup. A total of 58 strains of C. jejuni were ungroupable: 33 (13%) of the 259 hippurate-positive strains and 25 (44%) of the 57 hippurate-negative strains. All 121 strains of C. fetus tested were groupable as A, B, or A:B. The 14 conjugates used to define serogroups of C. jejuni, C. coli, and C. fetus reacted with the flagella but not the cells of other Campylobacter species and were negative on 256 other bacteria from 21 genera.
Pseudorabies virus nucleocapsid antigen for skin testing in swine
Authors: Scherba G , Turek J J , Gustafson D P
Published: January 2020

A procedure is described for the production of an effective pseudorabies virus skin test antigen consisting of nucleocapsids. The nucleocapsids were prepared by disruption of pseudorabies virus-infected pig kidney cells and nonionic detergent removal of the viral envelope. This preparation was noninfectious for pig kidney cell cultures and mice. Swine that had recovered from a pseudorabies virus infection and were injected intradermally with the nucleocapsid antigen developed a delayed type hypersensitivity response similar to that induced by inactivated virion antigen. The major advantage of the nucleocapsid antigen is that the occasional problem of seroconversion that is encountered with the virion antigen is avoided.
Analysis of
Neisseria gonorrhoeae
for In Situ β-Lactamase Production by Reagent-Impregnated Filter Paper Replica Methods
Authors: Bae Ben H. C. , Ledesma G. , Korzis J.
Published: January 2020

Reagent-impregnated filter paper replica methods are described which can be used to detect all penicillinase (β-lactamase)-producing
Neisseria gonorrhoeae
(PPNG) colonies in an entire population of
Neisseria gonorrhoeae
growing on primary isolation plates.
Rapid Determination of Novobiocin Resistance of Coagulase-Negative Staphylococci with the MS-2 System
Authors: Almeida Roy J. , Jorgensen James H.
Published: January 2020

Special novobiocin elution disks were prepared for testing of coagulase-negative staphylococci in the MS-2 system (Abbott Laboratories, Diagnostics Div., Irving, Tex.). The MS-2 system correctly classified 91.5% of 82 isolates as either susceptible or resistant to novobiocin in an average time of 5.8 h.
Age-related prevalence of complement-fixing antibody to Mycoplasma pneumoniae during an 8-year period
Authors: Pönkä A , Ukkonen P
Published: January 2020

We determined the age-related prevalence of complement-fixing antibody to Mycoplasma pneumoniae from the computerized laboratory results of our routine diagnostic department. The material consisted of about 58,000 sera from an 8-year period, 1971 to 1978. Among children less than 1 month old, the frequency of complement-fixing antibody of titers greater than or equal to 8 was low (23%), and no decrease representing loss of maternal antibody was seen thereafter. An unexpectedly early increase in the antibody prevalence up to 62% was seen by 6 months of age. The frequency of antibody was high among young children from the age of 4 months, in whom symptomatic infection due to M. pneumoniae is rare. The frequency of higher titers (greater than or equal to 32) and the geometric mean titer increased more slowly, coinciding with the known age distribution of symptomatic M. pneumoniae disease; the frequency of high titers and the geometric mean titer peaked at the age of 7 to 10 years. Two explanations for the high frequency of complement-fixing antibody to M. pneumoniae in young children are discussed. It may be due to an asymptomatic infection caused by M. pneumoniae or to a nonspecific stimulus by lipids of other organisms, plants, or tissues leading to production of antibodies crossreacting with M. pneumoniae, or it may be due to both of the above. During the study, two extensive epidemics due to M. pneumoniae occurred in Finland, but the prevalence of complement-fixing antibody bore no correlation with these peaks of occurrence.
Agglutination of Naegleria fowleri and Naegleria gruberi by antibodies in human serum
Authors: Reilly M F , Marciano-Cabral F , Bradley D W , Bradley S G
Published: January 2020

The capability of serum samples from 423 human subjects to agglutinate rounded cells of Naegleria fowleri nN68 was assessed. Sera from the umbilical cords of seven infants failed to agglutinate N. fowleri cells. The median agglutination titer was 1:4 for sera from children through age 4 years, 1:8 for sera from juveniles 5 to 15 years of age, and 1:16 for sera from subjects 15 to 30 years old. The agglutination titers of sera from older adults decreased to a median of 1:8 for the 40- to 60-year-old age group and to 1:4 for the 60- to 90-year-old subjects. Serum samples from young adults agglutinated rounded cells of both N. fowleri and N. gruberi. The agglutination activity for N. fowleri was removed by absorption with N. fowleri but not with N. gruberi. Conversely, agglutination activity for N. gruberi was removed by absorption with N. gruberi but not with N. fowleri. The agglutinating activity for N. fowleri was immunoglobulin M. Serum samples from children displayed markedly disparate capabilities to agglutinate N. fowleri and N. gruberi. Only rounded cells of N. fowleri or N. gruberi were reliably agglutinated by human serum samples. Live or paraformaldehyde-killed cells could be used in the assay, but live N. gruberi cells returned to the amoeboid form, and these agglutinated poorly.
Cytomegalovirus-specific cell-mediated immunity in lower-socioeconomic-class adolescent women with local cytomegalovirus infections
Authors: Faix R G , Zweig S E , Kummer J F , Moore D , Lang D J
Published: January 2020

The factors that regulate cytomegalovirus (CMV) excretion from the genitourinary tract are poorly understood. To assess the role of cell-mediated immunity in such excretion, a CMV-specific mononuclear blastogenesis assay was used to study a predominantly lower-socioeconomic-status population of 92 healthy nonpregnant adolescent women who also had CMV complement-fixing antibody titers and viral cultures of cervix, urine, saliva, and blood performed. Eighteen were studied more than once. No blood cultures were positive and no seroconversions were noted. There was no significant difference for frequency or degree of systemic CMV-specific blastogenesis between the 20 who were culture positive and the 41 who were seropositive but culture negative, although 40% of the culture-positive group and 27% of the seropositive, culture-negative group lacked CMV-specific blastogenesis. One of 31 seronegative subjects displayed CMV-specific blastogenesis. No systematic deficits were noted in any groups or individuals for E rosette number or mitogen response, though some isolated significant differences among groups for mitogen responses existed. Local CMV excretion in the study population was not related to systemic CMV-specific mononuclear blastogenesis.
Phenotypic markers associated with gastrointestinal Aeromonas hydrophila isolates from symptomatic children
Authors: Janda J M , Bottone E J , Skinner C V , Calcaterra D
Published: January 2020

Aeromonas hydrophila gastroenteritis was detected in 12 pediatric patients during a 5-month period. Chief complaints included bloody diarrhea, fever, vomiting, and abdominal pain. Severe symptoms in two patients necessitated hospitalization and supportive care. Phenotypic characteristics associated with enterotoxigenicity of A. hydrophila strains demonstrated that all 12 isolates were cytotoxic to HeLa cells and most were lysine decarboxylase positive (75%). A correlation existed between the presence of the five virulence-associated markers of two isolates of A. hydrophila and the severity of disease. Although the length and symptoms of gastroenteritis varied among all 12 patients, most had self-limiting diarrhea. The frequent occurrence of A. hydrophila gastroenteritis in pediatric patients warrants a greater appreciation of this agent as a significant cause of diarrhea, especially in summer.
Practical protocol for cytomegalovirus isolation: use of MRC-5 cell monolayers incubated for 2 weeks
Authors: Gregory W W , Menegus M A
Published: January 2020

Isolation of cytomegalovirus (CMV) in tissue culture is presently the most reliable means of proving active CMV infection. To improve both the cost-effectiveness and clinical usefulness of procedures for the isolation of CMV from fresh clinical specimens, we analyzed results obtained with standard isolation procedures and compared them with results obtained under different conditions. Cell monolayers from commercial sources were inoculated with fresh specimens and then were observed for a cytopathic effect typical of CMV. Of 1,375 specimens submitted over a 12-month period, 6.4% were CMV positive in WI-38 monolayers within 28 days after inoculation. The mean day of CMV detection for 45 urine, 13 cervical-vaginal, and 5 saliva specimens was 6.7 +/- 3.1 (mean +/- standard deviation), 9.9 +/- 3.3, and 7.7 +/- 3.3 days, respectively, and 92% were positive within 14 days. When 1,058 subsequent specimens were inoculated in parallel onto WI-38 and MRC-5 cell monolayers, 8.7% were positive for CMV. MRC-5 cells were significantly more sensitive than WI-38 cells: 98% of all positive specimens appropriate for comparison were detected in MRC-5 cultures, but only 85% were detected in WI-38 cells. Although 1 specimen was positive in WI-38 cells only, 38% of all isolates were positive earlier (16 specimens) or only (10 specimens) in MRC-5 cultures. Based on these data, we have developed a practical 2-week protocol for CMV isolation from fresh clinical specimens that includes the use of MRC-5 cell monolayers incubated at 36 degrees C.
Evaluation of the usefulness of counterimmunoelectrophoresis for diagnosis of Clostridium difficile-associated colitis in clinical specimens
Authors: Wu T C , Fung J C
Published: January 2020

Results of counterimmunoelectrophoresis (CIE) were compared with those of isolation of Clostridium difficile and assay for cytotoxicity in HeLa cells. On the basis of 471 stool specimens, CIE exhibited a sensitivity of 38% and a specificity of 88% as compared with the cytotoxin assay. The predictive value of a reactive CIE results is low (17%), whereas the predictive value of a nonreactive CIE result is significant (96%) and therefore warrants its use as a screening test. In addition, stool filtrates may nonspecifically precipitate with the C. difficile antitoxin in the CIE test. Such nonspecific reactions may be identified by simultaneous electrophoresis against nonimmune serum.
Enzyme-linked immunosorbent assay for detection of human antibodies to Salmonella typhi Vi antigen
Authors: Barrett T J , Blake P A , Brown S L , Hoffman K , Llort J M , Feeley J C
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies to Salmonella typhi Vi antigen in human serum, and the results were compared with those from a previously described hemagglutination assay (HA). The ELISA detected Vi antibodies at a titer of greater than or equal to 20 in 40 (52%) of 77 sera from typhoid fever patients, whereas the HA gave titers of greater than or equal to 20 in 35 (47%). Determination of titers of serum specimens from 170 persons without typhoid fever revealed Vi antibody titers of greater than or equal to 20 in 4 (2.3%) by the ELISA and 3 (1.7%) by the HA. Unlike the sensitized erythrocytes used in the HA, the ELISA reagents have a shelf life of greater than or equal to 1 year. The ELISA may be preferred by some laboratories, especially those already performing other ELISA tests.
Evidence for transient Staphylococcus epidermidis bacteremia in patients and in healthy humans
Authors: Zierdt C H
Published: January 2020

A new blood lysis-filtration culture technique revealed a high incidence of Staphylococcus epidermidis in the blood of patients and of healthy people. Of 2,004 blood cultures from patients, the blood lysis method grew S. epidermidis in 233 (11.6%), whereas a conventional two-bottle culture system grew this organism in 48 (2.4%). To determine the incidence deriving from the skin, 100 mock blood cultures by each technique were performed. The antecubital fossa was prepared as for a phlebotomy. The needle was inserted through the skin but not into the vein. Needles were cultured by conventional and lysis-filtration culture. A total of 1 conventional culture of 100 (1%) and 2 lysis-filtration cultures of 100 (2%) grew S. epidermidis. Of 100 lysis-filtration and conventional control cultures with broth in place of blood, no cultures were positive. Blood samples from 8 of 117 (6.8%) healthy blood donors were positive for S. epidermidis by lysis-filtration, whereas no matching conventional cultures were positive. Phage typing patterns of skin and blood strains from selected individuals were the same. S. epidermidis isolates were often concomitant with isolates of bona fide pathogens. I conclude that intermittent, transient, asymptomatic S. epidermidis bacteremia occurs frequently in patients and in healthy humans.
Prevalence of Eikenella corrodens in dental plaque
Authors: Goldstein E J , Tarenzi L A , Agyare E O , Berger J R
Published: January 2020

The prevalence of Eikenella corrodens in dental plaque and saliva samples of 282 volunteers was determined by a semiquantitative method with a selective medium. E. corrodens was recovered in 58.9% of plaque samples and 0.3% of saliva samples. This prevalence rate was not significantly altered (P greater than 0.05) by variables of sex, race, smoking habits, clinic attended by the patient, general health status, or age; however, patients 7 to 14 years old had a significantly higher prevalence rate (90.5%; P less than 0.05). E. corrodens should be considered as a potential pathogen in infections associated with and wounds inoculated by the human gingival flora.
Edwardsiella tarda isolated in Israel between 1961 and 1980
Authors: Sechter I , Shmilovitz M , Altmann G , Seligmann R , Kretzer B , Braunstein I , Gerichter C B
Published: January 2020

Edwardsiella tarda was isolated from patients, water tortoises (Clemmys caspica), and samples of water from Lake Kinnereth, the river Jordan, well water, and sewage water. Of the 53 isolates, 35 belonged to completely identified serotypes, among them 7 new ones. Fourteen cultures had O antigens, and one had an H antigen, different from those previously described. Three serotypes isolated from patients were also found in other sources: water tortoises, lake water, or both.
Typing of herpes simplex virus by an enzyme-linked immunosorbent assay with monoclonal antibodies
Authors: Nilheden E , Jeansson S , Vahlne A
Published: January 2020

We developed a method that uses monoclonal antibodies for typing herpes simplex virus type 1 and type 2 strains. Clinical isolates from GMK cells were seeded directly into a monolayer of GMK cells. After incubation overnight, monoclonal antibodies were added to the infected monolayer, and antibody binding was indicated by a peroxidase enzyme-linked immunosorbent assay. Using prototype strains and previously typed patient strains, we verified the specificity of this technique. This method is now used routinely for typing herpes simplex strains in our laboratory. We have also used this technique for specific staining of type 1 plaques in a mixture of type 1 and type 2 plaques. With this method it is possible to find a single type 1 plaque among several hundred type 2 plaques on a single petri dish. Infectious virus can also be recovered from stained, unfixed type 1 plaques.
Antigenic relationships between the enteroinvasive Escherichia coli O antigens O28ac, O112ac, O124, O136, O143, O144, O152, and O164 and Shigella O antigens
Authors: Cheasty T , Rowe B
Published: January 2020

Enteroinvasive Escherichia coli strains have been found in the following O serogroups: O28ac, O112ac, O124, O136, O143, O144, O152, and O164. The biochemical reactions of these enteroinvasive E. coli strains are often similar to those of Shigella strains, and identification may be difficult. The problem of differentiation and identification is further complicated by the sharing of antigenic components. The antigenic relationships between these O antigens and those of all the established and provisional Shigella serovars were examined. O antigen identity was demonstrated between E. coli O124 and Shigella dysenteriae 3 and between E. coli O152 and provisional Shigella serovar 3341:55. Reciprocal relationships were found between E. coli O112ac and S. dysenteriae 2, S. boydii 1, S. boydii 15, between E. coli O136 and S. dysenteriae 3, and between E. coli O164 and S. dysenteriae 3. A one-way relationship was demonstrated between E. coli O136 and S. boydii 1. The remaining enteroinvasive E. coli O serogroups showed no significant antigenic relationships with any Shigella serovar.
Establishment of minimum inhibitory concentrations of cefoperazone for control and reference anaerobic organisms
Authors: Zabransky R J , Randall E , Sutter V L , Birk R J , Westenfelder G , Emmerman J , Ghoneim A T
Published: January 2020

The minimum inhibitory concentrations of cefoperazone were determined in a collaborative study for eight National Committee for Clinical Laboratory Standards control and reference strains of anaerobic bacteria by agar and microdilution techniques with several types and sources of media. Recommended minimum inhibitory concentrations for the control strains, Bacteroides fragilis ATCC 25285 and Bacteroides thetaiotaomicron ATCC 29741, are 32 to 64 micrograms/ml and 64 micrograms/ml, respectively. Clostridium perfringens ATCC 13124 gave inconsistent results, and no value is recommended. Recommended values for reference strains are presented. Modification of media did not significantly change the minimum inhibitory concentrations.
A new serogroup (L) of Neisseria meningitidis
Authors: Ashton F E , Ryan A , Diena B , Jennings H J
Published: January 2020

A strain of neisseria meningitidis (LCDC 78189) isolated from the mother of a 3-year-old male with meningococcal meningitis was found to be antigenically distinct from the known serogroups A, B, C, D, H, I, K, X, Y, Z, 29E, and W135; it was designated serogroup L. Anti-78189 serum specifically agglutinated the homologous strain and three other strains which were isolated from the father and two other contacts of the child. Only those strains isolated from the contacts produced immunoprecipitates with the anti-78189 serum by the antiserum-agar method. A structurally unique capsular polysaccharide which was obtained from strain 78189 in a highly purified state was demonstrated to be the antigen responsible for the serological properties of the strain. The polysaccharide formed a precipitin band with the anti-78189 serum but not with the meningococcal grouping sera, and it was also able to absorb both the agglutinating and precipitating activity from the anti-78189 serum.