bluuebunny/crossref_metadata_2025 huggingface dataset:
Streptococcus faecium outbreak in a neonatal intensive care unit
Authors: Coudron P E , Mayhall C G , Facklam R R , Spadora A C , Lamb V A , Lybrand M R , Dalton H P
Published: January 2020

An outbreak of bacteremia and meningitis in a neonatal intensive care unit is described. Seven cases occurred in premature infants with severe underlying diseases. An epidemiological investigation failed to document the reservoir of the epidemic strain but suggested that its transmission among the infants was via the hands of hospital personnel. All patients had nasogastric tubes and multiple intravascular devices, and the portal of entry may have been either the gastrointestinal tract or the sites of the intravascular devices. Conventional biotyping of isolates failed to differentiate between isolates from infected patients and isolates recovered from prevalence surveys and from the environment. However, rapid identification systems (API-20S [Analytab Products, Plainview, N.Y.] and the AutoMicrobic system [Vitek Systems, Inc., Hazelwood, Mo.]) were able to distinguish isolates recovered from infected patients and hands of hospital personnel from isolates recovered during prevalence and environmental surveys and 29 isolates from widespread geographical areas. This is the first known report of a nosocomial neonatal outbreak of bacteremia and meningitis due to Streptococcus faecium; it underscores the importance of identifying streptococci to species level.
Evaluation of a rapid method to exclude the presence of certain enteric pathogens in stool specimens
Authors: Teti G , Burdash N M , Zamboni C , Fava C , Tomasello F , Mastroeni P
Published: January 2020

A new commercial method intended to exclude the presence of Salmonella spp., Shigella spp., and Yersinia enterocolitica and to presumptively identify Salmonella isolates within 2 h after primary isolation from stool specimens was evaluated. This system is marketed in Europe as API Z and in the United States as Rapid SST. The strip consists of five pairs of cupules for the screening of five lactose-negative colonies. The first cupule of each pair detects the presence of five enzymatic activities, whereas the second serves to maintain the strain for additional testing if necessary. A total of 197 fresh isolates from stool specimens and 217 stock cultures of Salmonella spp., Shigella spp., and Yersinia enterocolitica were tested, with the API 20E system as a reference method. In the stool specimens, 77.3% of the bacteria could be excluded from further workup for the presence of these organisms within 2 h. Over 97% of the stock strains and each of three fresh Salmonella isolates tested produced a reaction pattern corresponding to a correct presumptive identification. This reaction pattern was not produced by any isolate other than the Salmonella isolates. The API Z system can be used as a screen for the presence of Salmonella and Shigella spp. and can provide an accurate presumptive identification of Salmonella isolates within 2 h after primary isolation.
Automated determination of anti-streptolysin O antibodies by a kinetic hemolytic method
Authors: Ricci A , Arezzini C , Cocola F , Meiattini F
Published: January 2020

An automated method which uses oxidized Streptolysin O and determines the anti-Streptolysin O titer in whole blood without dilution of the sample is described. The addition of a reducing agent starts the hemolysis at an initial rate which is inversely proportional to the anti-Streptolysin O titer of the sample. An instrument (Taso-matic), designed to automatically execute all of the procedures, was used. Comparison between the described method and the classical hemolytic method gave the following linear regression slope y = 1.06x-13 (r = 0.974). Precision of the method at two different anti-Streptolysin O titers (200 and 500 IU), expressed as relative standard deviation, was 12.1 and 4.7% in the between-run procedure and 10.1 and 4.9% in the within-run procedure, respectively.
Outbreak of Salmonella typhimurium gastroenteritis due to an imported strain resistant to ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole in a nursery
Authors: Lamb V A , Mayhall C G , Spadora A C , Markowitz S M , Farmer J J , Dalton H P
Published: January 2020

An outbreak caused by a highly resistant strain of Salmonella typhimurium occurred in a nursery at a university medical center. The outbreak strain, which was resistant to ampicillin, chloramphenicol, and trimethoprim-sulfamethoxazole, was apparently imported from the Far East by a Cambodian refugee. The five patients involved had severe underlying diseases, and bacteremia and meningitis developed in one of these patients. The only reservoir identified was the gastrointestinal tracts of the infected patients, and infection was probably transmitted by the contaminated hands of hospital personnel. The outbreak was rapidly brought under control by isolating cases outside of the nursery and by instituting enteric precautions for infants who remained in the nursery. When compared by disk diffusion susceptibility tests with 353 strains of S. typhimurium tested at the Centers for Disease Control, the imported strain had a unique antibiogram. Bacteriophage typing of the strains revealed that all were untypable; this, in itself, was a good marker, because only 5 to 10% of S. typhimurium isolates in this country have this property. Agarose gel electrophoresis of isolates from the five patients revealed an identical plasmid banding pattern consisting of three large and three small plasmids. Highly resistant strains of S. typhimurium imported from the Far East may spread rapidly when introduced into a hospital nursery. Prompt institution of control measures may limit the outbreak and prevent systemic infections for which there are few effective therapeutic agents.
Cytomegalovirus in urine: detection of viral DNA by sandwich hybridization
Authors: Virtanen M , Syvänen A C , Oram J , Söderlund H , Ranki M
Published: January 2020

A cytomegalovirus (CMV)-specific sandwich hybridization test was constructed by using two adjacent BamHI DNA fragments of CMV DNA as reagents. The fragments were cloned into two different vectors. One of the recombinants was attached to the filter, and the other was the labeled probe. When present in the sample, CMV DNA mediated labeling of the filter by hybridizing to both the filter-bound DNA and the probe. The sandwich hybridization test was applied for the detection of CMV DNA from urine. DNA was released from virus by 2% Sarkosyl, concentrated by 2-butanol extraction and isopropanol precipitation, denatured, and finally subjected to the sandwich hybridization test. As a result, 70 to 90% of the original viral DNA could be recovered and demonstrated by the quantitative hybridization reaction. Urine could be stored at room temperature in Sarkosyl for at least 2 days without affecting the detectability of CMV. The clinical applicability of the test was evaluated by studying urine samples from four infants excreting CMV. Sandwich hybridization demonstrated the presence of CMV DNA in all of the specimens. These contained originally 10(5) to 10(8) CMV DNA molecules per ml.
Biotypes of serologically nontypable Haemophilus influenzae isolated from the middle ears and nasopharynges of patients with otitis media with effusion
Authors: DeMaria T F , Lim D J , Barnishan J , Ayers L W , Birck H G
Published: January 2020

A biotype analysis was performed on serologically nontypable Haemophilus influenzae isolates from the middle ear effusions and nasopharynges of 33 children with chronic otitis media with effusion (serous otitis media). Over 50% of the H. influenzae isolates from the middle ears belonged to biotype II. The incidence of beta-lactamase production was 17%. The results of this study indicate that future clinical or experimental investigations of chronic otitis media with effusion induced by serologically nontypable H. influenzae should focus on this clinically important biotype.
Enzyme-linked immunosorbent assays for Staphylococcus aureus exfoliative toxins A and B and some applications
Authors: Piémont Y , Haubensack M , Monteil H
Published: January 2020

Two enzyme-linked immunosorbent assays were developed for detection of staphylococcal exfoliative toxins A and B (ETA and ETB) with a double-antibody sandwich protocol. Antibodies against both toxins were purified by affinity chromatography from sheep antisera raised against purified ETA and ETB. These affinity-purified antibodies were free of detectable amounts of antibodies to other staphylococcal antigens and neutralized the actions of ETA and ETB. Alkaline phosphatase was conjugated to these antibodies. The enzyme-linked immunosorbent assay, which could detect at least 3 ng of ETA and ETB per ml, was used to quantitate the toxins in the culture supernatant fluids of staphylococcal strains. Thus, the kinetics of ETA and ETB synthesis and of ETA and ETB release into the supernatant fluids were determined; other determinations included the roles of carbon dioxide concentration, pH, glucose concentration, temperature, and agitation on the production of ETA and ETB.
Differentiation of Bacteroides ovatus and Bacteroides thetaiotaomicron by means of bacteriophage
Authors: Cooper S W , Szymczak E G , Jacobus N V , Tally F P
Published: January 2020

Two members of the Bacteroides fragilis group, B. ovatus and B. thetaiotaomicron, are difficult to distinguish by biochemical methods. They are currently identified on the basis of their variable ability to ferment salicin. We studied a method of identification for these two species by using cell lysis by bacteriophages. A total of 38 bacteriophages were used to distinguish the two species. Identification by bacteriophages was compared with species identification by prereduced anaerobically sterilized biochemical testing with salicin as the differentiating test. A total of 215 clinical isolates biochemically identified as B. ovatus or B. thetaiotaomicron were tested. A total of 100% of the strains identified as B. ovatus by bacteriophages produced strong acid in salicin (pH less than or equal to 5.4). However, 40% of the strains identified as B. thetaiotaomicron by bacteriophages also produced strong acid in salicin, and an additional 39% produced weak acid (pH 5.5 to 5.7). This study demonstrates that salicin fermentation is an inadequate test for the differentiation of B. ovatus and B. thetaiotaomicron.
Reproducibility of the MS-2 system for identification of members of the family Enterobacteriaceae: a collaborative study with blindly assigned reference stains
Authors: Yamane N , Kato H , Saitoh Y , Mukojima T , Matsuoka K
Published: January 2020

The reproducibility of identification and biochemical reactions for five different reference organisms of Enterobacteriaceae; Proteus vulgaris, Klebsiella pneumoniae, Escherichia coli, Serratia marcescens, and Enterobacter cloacae, were evaluated using the updated MS-2 system software (Abbott Laboratories, Diagnostic Division, Irving, Tex.) in a collaborative study involving 11 laboratories. When a total of 220 randomly coded test organisms were blindly examined, the MS-2 system correctly identified 92.7 and 86.8% for over 80 and 90% probability identification, respectively. Four organisms, P. vulgaris, K. pneumoniae, E. coli, and S. marcescens, were correctly identified in all laboratories with high probability, but 9 of 44 tests of Enterobacter cloacae resulted in misidentifications or low-likelihood (less than 80%) identifications. Accuracy was directly related to level of experience and familiarity with the MS-2 system in the individual laboratories. Biochemical reactions varied among the identification trials, especially in the identification of S. marcescens and Enterobacter cloacae. Among a total of 44 subcultures for each organism, 10 different biochemical patterns for P. vulgaris, 6 for K. pneumoniae, 9 for E. coli, 15 for S. marcescens, and 14 for Enterobacter cloacae were obtained. The results indicate that the MS-2 system performs with high accuracy and reproducibility in identifying Enterobacteriaceae, except for Enterobacter cloacae.
Rapid differentiation of enterotoxigenic Escherichia coli that produce heat-stable and heat-labile toxins by frequency-pulsed electron capture gas-liquid chromatography analysis of diarrheal stool specimens
Authors: Brooks J B , Basta M T , el Kholy A M , Moss C W
Published: January 2020

Thirty-three stool specimens from infants in the village of Tamooh near Cairo, Egypt, were studied by frequency-pulsed electron capture gas-liquid chromatography (FPEC-GLC). In 13 of the diarrheal cases, the suspected causative agent isolated was Escherichia coli which produced heat-stable toxin (ST), and in 10 other cases E. coli that produced heat-labile toxin (LT) were isolated. Ten control stool samples, collected from infants from whom no pathogenic organisms were isolated, were analyzed at the same time. Comparisons also were made against healthy control stools from individuals in the United States who had been previously analyzed by FPEC-GLC (Brooks et al., J. Clin. Microbiol. 20:549-560, 1984). The stools were suspended in water and centrifuged, and the supernatant was extracted with organic solvents and derivatized to form electron-capturing derivatives of carboxylic acids, hydroxy acids, alcohols, and amines. Results from the study showed distinct differences among the FPEC-GLC profiles of E. coli ST-positive stools, of E. coli LT-positive stools, and of the control stool samples. An unidentified compound appearing in the ether-soluble hydroxy acid fraction from E. coli ST-positive stools was tentatively identified by mass spectrometry as 6-methoxy-2-hydroxyhexanoic acid. 6-Methoxy-2-hydroxyhexanoic acid was found in all stools that contained E. coli ST but was not present either in stools from which E. coli LT was isolated or in control samples. 6-Methoxy-2-hydroxyhexanoic acid may prove to be an important marker for use in the identification of E. coli ST. In addition to 6-methoxy-2-hydroxyhexanoic acid, the carboxylic acid, alcohol, and amine FPEC-GLC profiles obtained from stools were very different between these two organisms. The data indicate that FPEC-GLC analysis of diarrheal stool specimens might be a rapid way to distinguish diarrhea caused by E. coli ST, E. coli LT, Clostridium difficile, and rotavirus.
Antibody responses to capsular polysaccharide, lipopolysaccharide, and outer membrane in adults infected with Haemophilus influenzae type b
Authors: Lagergård T , Nylén O , Sandberg T , Trollfors B
Published: January 2020

The antibody response to Haemophilus influenzae type b antigens as capsular polysaccharide, lipopolysaccharide, and outer membrane components was studied by enzyme-linked immunosorbent assay in 10 adults infected with this bacterium. Almost all patients had detectable amounts of immunoglobulins G and M antibodies specific to capsular polysaccharide, lipopolysaccharide, or outer membrane in their first serum sample. A significant antibody response in one or more antibody subclasses to capsular polysaccharide, lipopolysaccharide, and outer membrane was noted in 9 of 10, 7 of 10, and 7 of 10 patients, respectively. The occurrence of H. influenzae type b infections in adults cannot be completely explained either by the absence of antibodies against H. influenzae type b in their serum or by the failure to develop specific antibodies to the capsule or certain cell wall components even if these factors probably are important in some cases, e.g., in one patient who was splenectomized.
Evaluation of a C-reactive protein latex agglutination detection test with sera from patients with sexually transmitted diseases
Authors: Schalla W O , Arko R J , Thompson S E
Published: January 2020

A total of 149 sera, including 79 pre- and posttreatment sera from 33 patients with disseminated gonococcal infections, 18 from patients with uncomplicated gonococcal infections, 6 from patients with pelvic inflammatory disease, 4 from patients with genital Chlamydia trachomatis infections, and 42 from normal volunteers, were examined for C-reactive protein with a latex agglutination C-reactive protein detection kit (Difco Laboratories, Detroit, Mich.). Results were quantitated with LC-Partigen C-reactive protein radial immuno-diffusion plates (Calbiochem-Behring, La Jolla, Calif.). Positive latex agglutination results were observed in all of the pretreatment sera and some of the posttreatment sera of patients with disseminated gonococcal infections and in two sera from patients with pelvic inflammatory disease, which corresponded to quantitative C-reactive protein levels in the radial immunodiffusion plates. C-reactive protein levels were not detectable in the serum samples from normal volunteers or patients with uncomplicated gonococcal infections or genital chlamydial infections. Positive latex agglutination occurred as early as 20 s in sera with high C-reactive protein levels, and all positive results were observed within 90 s of the 3-min test limit. Positive latex test results were obtained with C-reactive protein levels as low as 1 mg/dl (10 micrograms/ml).
Correlation of leukocyte esterase activity and bacterial isolation from body fluids
Authors: Smalley D L , Bradley M E
Published: January 2020

We evaluated 230 body fluid samples, of which 131 were peritoneal effluents and 99 were other body fluids. Of these, 63 dialysates were culture positive, and 54 (85.7%) of these 63 were leukocyte esterase positive. Of 99 other body fluids, 8 were both culture positive and leukocyte esterase positive.
Simple spot test for rapid detection of urease activity
Authors: Qadri S M , Zubairi S , Hawley H P , Ramirez E G
Published: January 2020

A spot test for the detection of urease activity was developed and evaluated with 761 strains of gram-negative bacteria. The test was compared with the conventional Christensen urea agar slants and urease test on the Vitek Enterobacteriaceae card (Vitek Systems, Inc., St. Ana, Mo.). Of the 348 strains of the Proteus-Providencia-Morganella group that were urease positive, 327 (94%) yielded positive results within 1 min, and all strains yielded positive results within 2 min. All these organisms also gave a positive urease reaction on the Vitek Enterobacteriaceae card test within 5 h and on the Christensen urea agar slants in 4 to 48 h. All the bacteria that did not hydrolyze urea by these two tests also remained negative by the spot test.
Evaluation of the Strep-A-Fluor identification method for group A streptococci
Authors: Wasilauskas B L , Hampton K D
Published: January 2020

Strep-A-Fluor (Bio Spec, Inc., Dublin, Calif.) is a new test designed for the rapid identification of group A streptococci. A filter paper strip impregnated with a synthetic substrate is used to detect a specific aminopeptidase present in group A streptococci by UV fluorescence. In a blind study, 305 beta-hemolytic streptococcal isolates were correctly categorized as group A or non-group A.
Evaluation of fluorescent-antibody tests as a means of confirming infant botulism
Authors: Glasby C , Hatheway C L
Published: January 2020

Fluorescent-antibody techniques were evaluated for confirming infant botulism. Seventy-seven stool specimens from suspected cases were examined. All 34 specimens containing viable Clostridium botulinum at time of study gave positive results (29 on direct smears and 34 on enrichments). Two false-positive reactions were observed.
Value of blood agar for primary plating and clinical implication of simultaneous isolation of Aeromonas hydrophila and Aeromonas caviae from a patient with gastroenteritis
Authors: Janda J M , Dixon A , Raucher B , Clark R B , Bottone E J
Published: January 2020

The simultaneous recovery of Aeromonas hydrophila and Aeromonas caviae from the stool of a 49-year-old woman with watery diarrhea was facilitated through the use of a blood agar medium which detected the hemolytic capability of A. hydrophila. In vitro phenotypic tests support the conclusion that only the A. hydrophila isolate was clinically significant.
Nosocomial pneumonia caused by a glucose-metabolizing strain of Neisseria cinerea
Authors: Boyce J M , Taylor M R , Mitchell E B , Knapp J S
Published: January 2020

We describe what appears to be the first reported case of nosocomial pneumonia caused by Neisseria cinerea. The isolate metabolized glucose when tested in BACTEC Neisseria Differentiation Kits (Johnston Laboratories), but did not produce detectable acid in cystine-Trypticase (BBL Microbiology Systems) agar medium or in modified oxidation-fermentation medium. Clinical laboratories that rely on the BACTEC method for differentiation of pathogenic neisseriae should be aware of the fact that N. cinerea may mimic N. gonorrhoeae when tested in BACTEC Neisseria Differentiation kits. The ability of N. cinerea to grow well on tryptic soy and Mueller-Hinton agars and its inability to grow on modified Thayer-Martin medium are characteristics which help to distinguish N. cinerea from N. gonorrhoeae.
Comparison of PRAS II, RapID ANA, and API 20A systems for identification of anaerobic bacteria
Authors: Karachewski N O , Busch E L , Wells C L
Published: January 2020

This study evaluated the PRAS II, RapID ANA, and API 20A systems for the identification of anaerobic bacteria. A total of 80 isolates (68 fresh clinical isolates and 12 stock cultures) were examined and included 25 Bacteriodes spp., 7 Fusobacterium spp., 12 Clostridium spp., 2 Veillonella spp., 16 gram-positive cocci, and 18 gram-positive nonsporeforming bacilli. All isolates were initially identified by the procedures outlined in Holdeman et al. (ed.), Anaerobe Laboratory Manual, Virginia Polytechnic Institute and State University, Blacksburg, Va., 1977; identifications from the PRAS II, RapID ANA, and API 20A systems were compared with these initial identifications. If no supplemental tests were required, the RapID ANA and API 20A systems had incubation times of 4 and 24 h, respectively; the PRAS II system generally required 2 to 5 days of incubation, depending on the growth rate of the isolate. PRAS II identified 74% correct to species level, 14% correct to genus only, and 6% incorrect; 6% could not be identified. PRAS II data were reevaluated according to a revised data base that was provided after completion of the study; PRAS II (revised) identified 82% correct to species, 12% correct to genus only, and 6% incorrect. RapID ANA identified 62% correct to the species level, 28% correct to genus only, and 10% incorrect. API 20A identified 71% correct to the species level, 10% correct to genus only, and 3% incorrect; 16% could not identified. The API 20A is a more established system for identification of anaerobic bacteria; PRAS II and RapID ANA appear to be promising new methods for the identification of anaerobic bacteria.
Addition of three new serotypes of Shigella boydii to the Shigella schema
Authors: Wathen-Grady H G , Davis B R , Morris G K
Published: January 2020

No new serotypes have been added to the Shigella schema since 1958, although several provisional serotypes have been described. We conducted biochemical and serological studies on three provisional Shigella boydii serotypes. Four strains of serotype 2710-54 from four widely separated countries, 7 strains of serotype 3615-53 from three different countries, and 31 strains of serotype 1344-78 (E10163) from six different countries were included. Reactions of all three serotypes were consistent with those of S. boydii. On the basis of these results and other published research, we propose that these three provisional serotypes be admitted to the Shigella schema as S. boydii 16, 17, and 18.
Enzyme immunoassay of teichoic acids from Listeria monocytogenes
Authors: Kamisango K , Nagaoka M , Fujii H , Azuma I
Published: January 2020

Amino groups were introduced into Listeria monocytogenes teichoic acids by reductive amination, and the product was coupled to biotin. Teichoic acids were assayed by their binding to specific antibody adsorbed to a solid phase, followed by detection of the antigen-antibody complex by horseradish peroxidase-avidin. Less than 20 ng of teichoic acid was detectable.
Isolation and characterization of six new genome types of human adenovirus types 1 and 2
Authors: Fife K H , Ashley R , Corey L
Published: January 2020

Several of the 41 types of human adenovirus have been divided into genome types based on aberrant restriction endonuclease digestion patterns of viral DNA. In the process of screening a large number of clinical adenovirus isolates by restriction endonuclease digestion of viral DNA, we have identified nine isolates from eight patients which were identified by neutralization as adenovirus type 1 or 2 but had aberrant cleavage patterns. Cleavage sites for the enzymes SmaI, EcoRI, HindIII, KpnI, and HpaI were mapped. The variants could be placed in six distinct groups based on cleavage patterns. The designation genome types 1a, 1b, 1c, 2b, 2c, and 2d are proposed for these isolates.
Bacterial chromosomal restriction endonuclease analysis of the homology of Bacteroides species
Authors: Bradbury W C , Murray R G , Mancini C , Morris V L
Published: January 2020

Chromosomal DNAs of selected Bacteroides organisms whose relatedness had been previously determined by "conventional" filter-annealing studies (J. L. Johnson, Int. J. Syst. Bacteriol. 28:245, 1978) were further analyzed by restriction endonuclease analysis coupled with the Southern hybridization procedure (E. M. Southern, J. Mol. Biol. 98:503, 1975). By comparing their EcoRI restriction fragment patterns in agarose gel electrophoresis, each Bacteroides strain could be clearly differentiated. As a simple and direct means for comparison purposes this method was particularly useful for differentiating genetically similar organisms such as Bacteroides strains of the same species which shared greater than 75% homology. In contrast, bacterial chromosomal restriction endonuclease analysis in conjunction with Southern hybridizations was most effectively used to determine the significance of low levels of homology (less than 24%) as this technique provided additional information on the nature and relative distribution of that homology when the areas of homology were displayed as reproducible bands in autoradiograms.
Enzyme immunoassay for evaluation of Toxoplasma gondii growth in tissue culture
Authors: Merli A , Canessa A , Melioli G
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) for the evaluation of growth kinetics of Toxoplasma gondii in tissue cultures was developed. Tissue culture microplates (96 wells) were seeded with Vero cells, infected with a virulent strain of T. gondii, and incubated for different time periods. The ELISA was performed with anti-T. gondii antibodies on the infected cells. The method was simple, rapid, and accurate, and very good correlations between the ELISA results and the percentage of T. gondii rosettes in infected cells, the number of free T. gondii in the supernatant, and the amount of T. gondii antigen in the supernatant were observed.
Demonstration of a common antigen in sonicated cells for identification of Vibrio vulnificus serotypes
Authors: Nishibuchi M , Seidler R J
Published: January 2020

Ouchterlony immunodiffusion of sonicated Vibrio vulnificus cells illustrated a single major precipitation line with antiserum prepared from whole cells of the same species. Antigenic analysis by two-dimensional immunoelectrophoresis verified the presence of a single dominant precipitation line. Tandem two-dimensional immunoelectrophoretic analyses of V. vulnificus antigens from various strains revealed one fused precipitation line of identity. No fused precipitation lines were seen with other Vibrio species tested. This dominant antigen, designated VVA, was not dialyzable, lost antigenicity by heating at 100 degrees C but not at 70 degrees C, and was precipitated by 70%, but not by 50%, saturated ammonium sulfate. VVA was not present in concentrated (20X) spent culture medium. VVA is possibly an intracellular protein specific to V. vulnificus species and may be useful in the serological identification of this important human pathogen.
Use of a sodium polyanetholesulfonate disk for the identification of Gardnerella vaginalis
Authors: Reimer L G , Reller L B
Published: January 2020

Several methods have been previously suggested for the presumptive identification of Gardnerella vaginalis in clinical laboratories, but none is entirely satisfactory. We previously found that sodium polyanetholesulfonate (SPS) inhibits G. vaginalis in blood culture media. We compared susceptibility to an SPS-containing paper disk with beta-hemolysis on human blood agar, hippurate hydrolysis, and inhibition by alpha-hemolytic streptococci for identification of 62 previously confirmed G. vaginalis strains. All strains were positive by SPS disk and alpha-hemolytic streptococcus inhibition, 78% were positive by beta-hemolysis, and 81% were positive by hippurate hydrolysis. Although positive reactions occurred with SPS disk and alpha-hemolytic streptococcus tests for 5 and 9 of 84 other bacteria tested, respectively, none of these bacteria were positive for both tests. We conclude that a combination of SPS disk susceptibility and alpha-hemolytic streptococcus inhibition provides excellent identification of G. vaginalis when performed by the methods suggested.
Comparison of processing techniques for detection of Pneumocystis carinii cysts in open-lung biopsy specimens
Authors: Gay J D , Smith T F , Ilstrup D M
Published: January 2020

Methenamine silver stain was used to compare the number of cysts of Pneumocystis carinii contained in lung concentrate smears of homogenized lung tissue with the number in impression smears. Results were also compared with histopathological examination of methenamine silver-stained paraffin-embedded sections. Of slides from 23 preparations, a greater number of cysts were contained in concentrate smears than in impressions (P less than 0.001). In four preparations, cysts were noted in concentrate smears only. All concentrate smears were positive, whereas 11 of the 23 histopathological sections were negative (P less than 0.01). The ability to detect the cyst phase of P. carinii in lung tissue is enhanced by the use of concentrate smears.
Comparative evaluation of commercial rubella virus antibody kits
Authors: Wittenburg R A , Roberts M A , Elliott L B , Little L M
Published: January 2020

Comparative evaluation of six commercial rubella virus antibody kits is described. A set of serum samples skewed toward nonreactive and low-positive rubella antibody levels was used because the goal of an immunity screening test is the detection of susceptible individuals. The M. A. Bioproducts Rubastat and the Hynson, Westcott and Dunning Rubascan were superior in achieving high sensitivity, specificity, and reproducibility. They also were the easiest and fastest tests to perform, and they require equipment available in most laboratories.
Selective isolation and rapid identification of Clostridium botulinum types A and B by toxin detection
Authors: Dezfulian M , Bartlett J G
Published: January 2020

A simple procedure for rapid identification of Clostridium botulinum type A and B colonies from cultures and stool samples from infants with botulism was devised. The stool samples were directly streaked on C. botulinum isolation medium containing selective inhibitory agents. Typical lipase-positive colonies that appeared within 24 to 48 h were examined for the presence of botulinal toxin by the enzyme-linked immunosorbent assay and conventional mouse toxicity test. The amount of toxin associated with 48-h colonies of stock strains was comparable to that of 96-h broth culture. The quantity of toxin present in a single colony or combination of two was shown to be sufficient for toxin detection by the enzyme-linked immunosorbent assay. Of 42 additional stock strains tested in this manner, 41 (97.5%) were identified as toxigenic C. botulinum type A or B. The remaining one strain also proved to be toxigenic when it was tested as a concentrated cell suspension. This procedure should prove useful for large-scale serological screening of food and clinical specimens.
Leminorella, a new genus of Enterobacteriaceae: identification of Leminorella grimontii sp. nov. and Leminorella richardii sp. nov. found in clinical specimens
Authors: Hickman-Brenner F W , Vohra M P , Huntley-Carter G P , Fanning G R , Lowery V A , Brenner D J , Farmer J J
Published: January 2020

Leminorella is proposed as a new genus for the group of Enterobacteriaceae formerly known as Enteric Group 57. Strains of Leminorella gave positive tests for H2S production, acid production from L-arabinose and D-xylose, and tyrosine clearing; they were negative for indole production, Voges-Proskauer, urea hydrolysis, phenylalanine deaminase, motility, gelatin liquefaction, lysine and ornithine decarboxylases, arginine dihydrolase, growth in KCN, and acid production from adonitol, D-arabitol, cellobiose, erythritol, D-galactose, myo-inositol, lactose, maltose, D-mannitol, D-mannose, melibiose, alpha-CH3-glucoside, raffinose, L-rhamnose, salicin, D-sorbitol, sucrose, and trehalose. By DNA hybridization, strains of Leminorella were only 3 to 16% related to other Enterobacteriaceae and were divided into three groups. Leminorella grimontii is proposed as the type species for the genus and strain CDC 1944-81, ATCC 33999, is designated as the type strain. There were four strains of L. grimontii from stool specimens and two from urine specimens. L. richardii is proposed as the name for the second species (type strain, CDC 0978-82, ATCC 33998). All four L. richardii strains were from stool specimens. L. grimontii can be distinguished from L. richardii because it produces gas from glucose (100%) and acid from dulcitol (83%) and is methyl red positive (100%). One strain, CDC 3346-72, was more related to L. grimontii by DNA hybridization than to L. richardii, but the lower relatedness to both of these species indicated that it may be a third species. Biochemically it could not be distinguished from L. grimontii. All Leminorella strains were resistant (no zone of inhibition) to ampicillin, carbenicillin, and cephalothin. Some of the Leminorella strains were sent to us for Salmonella serotyping, and two reacted weakly in Salmonella antisera. The clinical significance of Leminorella is unknown.
Staphylococcus simulans septicemia in a patient with chronic osteomyelitis and pyarthrosis
Authors: Males B M , Bartholomew W R , Amsterdam D
Published: January 2020

Staphylococcus simulans was identified as the etiological agent of osteomyelitis and septic arthritis in an adult male who had sustained a fracture of the fibula and syndesmosis separation which required the installation of orthopedic hardware. Identifying characteristics and antibiograms for this organism, recovered from blood, wound exudate, and deep tissue samples, were determined. Recent evidence has linked slime production (adherence to smooth surfaces) by coagulase-negative staphylococci to infections by these organisms at sites where foreign bodies had been inserted. Tests for adherence showed this S. simulans strain to be a strong slime producer. This is the first reported case of osteomyelitis and septicemia due to S. simulans.
Actinomyces naeslundii as an agent of pelvic actinomycosis in the presence of an intrauterine device
Authors: Bonnez W , Lattimer G , Mohanraj N A , Johnson T H
Published: January 2020

Actinomyces naeslundii is a saprophyte, sometimes a pathogen, of the human oral cavity. Very few extra-oral infections related to this agent have been described. We report the first instance of A. naeslundii as an etiological agent of pelvic actinomycosis in a user of an intrauterine device, an infection so far exclusively attributed to Actinomyces israelii.
Pseudomonas pickettii as a cause of pseudobacteremia
Authors: Verschraegen G , Claeys G , Meeus G , Delanghe M
Published: January 2020

An outbreak of pseudobacteremia caused by Pseudomonas pickettii biovariant 1 is reported. The common source was the aqueous chlorhexidine solution prepared by the hospital pharmacy. The contamination problem caused by the antiseptic solution was eventually solved by a series of preventive measures.
Non-O1 Vibrio cholerae in Thailand: homology with cloned cholera toxin genes
Authors: Hanchalay S , Seriwatana J , Echeverria P , Holmgren J , Tirapat C , Moseley S L , Taylor D N
Published: January 2020

We examined 281 non-O1 Vibrio cholerae isolates from Thailand for homology with genes coding for cholera toxin. Five isolates from environmental sources were homologous with the cholera toxin gene probe and produced both the A and B subunits of cholera toxin.
Detection of Mycoplasma hominis septicemia by radiometric blood culture
Authors: Smaron M F , Boonlayangoor S , Zierdt C H
Published: January 2020

The ease with which Mycoplasma hominis can be recovered and the frequency of its occurrence in septicemia may not be fully appreciated. We detected the growth of M. hominis radiometrically with an automated blood culture instrument. The organism grew in both aerobic and anaerobic culture media, but the cultures were not visibly positive. It was necessary to stain the cultures with acridine orange to visualize M. hominis and to subculture them on Columbia base sheep blood agar to confirm the positive growth index indicated by the instrument. Sodium polyanetholesulfonate inhibited the growth of M. hominis and is not recommended for use as the anticoagulant when blood is cultured for Mycoplasma spp.
Improved recovery of group A beta-hemolytic streptococci with a new selective medium
Authors: Carlson J R , Merz W G , Hansen B E , Ruth S , Moore D G
Published: January 2020

A new selective group A streptococcus agar (ssA) (BBL Microbiology Systems, Cockeysville, Md.) was evaluated for the recovery and direct plate identification of group A beta-hemolytic streptococci (GABHS) in comparison with sheep blood agar (SBA). A total of 265 GABHS were recovered from 1,116 throat swab specimens on both media, with detection rates of 98.9 and 91.7% for ssA and SBA, respectively. Primary bacitracin disk susceptibility tests were performed on 549 specimens, and presumptive identifications were possible for 81.4% of the GABHS on ssA, as compared with 44.2% on SBA. All of the 120 GABHS recovered from another 567 specimens were identified by a coagglutination method after overnight incubation on ssA, whereas only 70.2% were identified at this interval on SBA. The major advantage of the ssA was the inhibition of normal respiratory flora, permitting improved recovery and rapid identification of GABHS.
Recognition of Staphylococcus saprophyticus in urine cultures by screening colonies for production of phosphatase
Authors: Pickett D A , Welch D F
Published: January 2020

Phenolphthalein diphosphate was incorporated into a primary blood agar medium for use in performing quantitative urine cultures. Phosphatase-negative staphylococci, such as Staphylococcus saprophyticus, were differentiated from phosphatase-positive species, such as Staphylococcus epidermidis, by spot testing colonies on filter paper saturated with 1 N NaOH. Phosphatase-positive colonies turned pink within seconds, and phosphatase-negative colonies showed no color. None of 55 S. saprophyticus isolates showed production of phosphatase on this medium. Of 193 consecutive coagulase-negative staphylococci isolated from the urine of 190 adolescent female patients, 84% were phosphatase positive, non-S. saprophyticus species; 16% were phosphatase-negative and indicated S. saprophyticus (22), Staphylococcus haemolyticus (4), Staphylococcus simulans (2), Staphylococcus warneri (1), and Staphylococcus hominis (1). Phosphatase activity was variable in the other flora encountered in the urine cultures. Mixtures of phosphatase-positive and -negative organisms did not cause false-positive reactions.
Pseudomonas mesophilica infections in humans
Authors: Smith S M , Eng R H , Forrester C
Published: January 2020

Reported here is the case of a patient with metastatic adenocarcinoma of the lung who had bacteremia involving Pseudomonas mesophilica. Of the common laboratory media tested at 35 degrees C, buffered charcoal yeast extract agar and nutrient agar provided the best growth; however, other media supported growth at lower temperatures. Since blood cultures are routinely subcultured onto chocolate agar and then incubated at 35 degrees C, awareness of the characteristics of P. mesophilica and of the isolation techniques as outlined may enhance the recovery of this and related bacterial species.
Decreased oxygen radical generation by neutrophils from patients with measles presumably owing to activation of suppressor T lymphocytes
Authors: Niwa Y , Sakane T , Somiya K , Miyachi Y
Published: January 2020

The capacity for the generation of oxygen radicals by polymorphonuclear leukocytes (PMNs) was assessed in 29 patients with measles and in control groups. Patients with secondary bacterial infections showed a significantly decreased generation of oxygen radicals; this abnormality did not persist for more than 2 months after disease onset. Normal PMNs incubated with T lymphocytes from these measles patients generated significantly fewer oxygen radicals than those incubated with T cells from either control group. However, normal PMNs incubated with non-T lymphocytes from these measles patients produced normal oxygen radical levels. In addition, irradiation above 1,500 rads of T lymphocytes abrogated the suppressive effect of T cells on PMNs. On the other hand, these abnormal findings were not observed in patients with measles but without secondary bacterial infections. The secondary bacterial infections sometimes seen in measles patients may result from a decrease in oxygen radical generation, presumably induced by the suppressive activity of the T lymphocytes of the patients.
Rapid diagnosis of acute mumps infection by a direct immunoglobulin M antibody capture enzyme immunoassay with labeled antigen
Authors: Gut J P , Spiess C , Schmitt S , Kirn A
Published: January 2020

A new immunoglobulin M (IgM) antibody capture enzyme immunoassay with peroxidase-labeled mumps antigen (dMACEIA) is described, and its suitability for practical diagnosis of acute mumps infection is evaluated. All 54 patients with proven mumps infection that were tested showed mumps-specific IgM antibodies. On the other hand, no specific IgM antibodies were present in 16 cases of suspected mumps that could not be confirmed by classical complement fixation serology, and IgM mumps virus antibodies could be detected neither in the sera of 100 healthy individuals nor in those of 16 patients positive for rheumatoid factor. In all, 22 children with acute respiratory illness caused by parainfluenza virus and 44 patients with infections due to other viruses showed no IgM response in mumps dMACEIA. The particular characteristic in which complement fixation antibodies against mumps nucleocapsids appear before and disappear earlier than antibodies to the enveloped mumps virus could not be demonstrated in the dMACEIA. In an extensive epidemic of mumps virus infection, the dMACEIA gave a clear diagnosis of mumps infection in 200 out of 371 suspected cases. By day 2 of the illness, 71% of the patients had detectable IgM, and by day 3, all of them had detectable IgM. In 99% of the cases, dMACEIA gave a positive result in the first available serum specimens, most of which were negative for complement fixation antibodies. A positive but only moderate correlation was thus observed between the two serological procedures. IgM antibodies persisted for at least 6 weeks. The dMACEIA, performed in 3 h, offers a reliable, simple, and rapid alternative to routine methods for detection of acute mumps infection.
Comparison of genomic homologies in the coxsackievirus B group by use of cDNA:RNA dot-blot hybridization
Authors: Tracy S
Published: January 2020

Radioactive cDNAs to coxsackievirus B (CB) genomic RNAs were used to probe genomic homologies between RNAs of the CB group serotypes. CB5 shared approximately equivalent homology with the other five CB serotypes. CB1, CB2, and CB3 hybridized preferentially to CB5 in turn, suggesting that these three have diverged in one direction from CB5. CB4 and CB6 were more related to each other than to the other serotypes at a lower criterion, but at a more stringent criterion they did not demonstrate apparent homology with each other. This suggests that they have diverged from CB5 as well but perhaps earlier in time or at a greater rate than did CB1, CB2, or CB3. Some potential applications of these relationships to clinical CB detection assays based on nucleic acid hybridization are discussed.
Four-step enzyme-linked immunosorbent assay for detection of Treponema pallidum antibody
Authors: Farshy C E , Hunter E F , Helsel L O , Larsen S A
Published: January 2020

Further studies of a four-step enzyme-linked immunosorbent assay procedure to detect Treponema pallidum antibody are described. High-titered antibody, produced in rabbits by intravenous injection of T. pallidum, was used to coat polyvinyl chloride microtiter plates. To these plates a known concentration of T. pallidum was added, followed in successive steps by serial dilutions of human sera and appropriately diluted peroxidase-labeled anti-human immunoglobulin G antibody. O-Phenylenediamine was the substrate. A total of 340 sera were obtained from the DeKalb County Sexually Transmitted Diseases Clinic, Atlanta, Ga., and examined within 3 days of receipt. Ninety-six percent test agreement between the enzyme-linked immunosorbent assay and the fluorescent treponemal antibody absorption-double staining test was obtained. A total of 372 additional sera stored at -20 degrees C were examined. The overall sensitivity of the enzyme-linked immunosorbent assay with sera from patients with various stages of syphilis was 96%. With sera from uninfected individuals, the specificity of the enzyme-linked immunosorbent assay was 95%. No antigen instability was noted with the two antigen preparations used during this evaluation.
Evaluation of a rapid method for the detection of streptococcal group A antigen directly from throat swabs
Authors: Venezia R A , Ryan A , Alward S , Kostun W A
Published: January 2020

Throat swabs from 196 pediatric patients were processed by a direct extraction-latex agglutination method (Group A Strep Direct Antigen Identification Test [DAI]) that detects group A streptococci in the specimen. The method requires a 45-min enzymatic extraction period at 37 degrees C and a 4-min reaction period with antibody-linked latex particles. The results were compared with those of the culture and fluorescent antibody methods and the clinical presentation of the patient for pharyngitis. Ninety-three percent of the specimens resulted in agreement by all tests, and 28% were culture positive for group A streptococci. Compared with the culture method, the DAI had a sensitivity and a specificity of 83% and 99%, respectively. The positive predictive values were 98% versus the culture method and 93% versus the fluorescent antibody method, whereas the negative predictive values were 94% versus both other methods. Of the 14 discrepant results when both clinical presentation of an acute pharyngitis and the test results were compared, the culture method provided the best correlation. An additional 64 specimens were processed by the DAI and another direct extraction-latex agglutination method (Culturette Ten-Minute Group A Strep ID Test), and the results were compared with those of the culture method. This group had a 40.6% culture isolation rate for group A streptococci. The sensitivity and specificity of the DAI and Strep ID methods versus the culture method were 81 and 100%, and 77 and 97%, respectively. These results indicate that the DAI is accurate for diagnosing group A streptococcal pharyngitis directly from throat swabs. However, negative results in the presence of a symptomatic patient must be confirmed by standard culture techniques.