bluuebunny/crossref_metadata_2025 huggingface dataset:
Variation in penicillin-binding protein patterns of penicillin-resistant clinical isolates of pneumococci
Authors: Markiewicz Z , Tomasz A
Published: January 2020

A large number of pneumococcal isolates (over 80 strains) from a variety of geographic locales and representing a spectrum of resistance levels from a penicillin MIC of 0.003 microgram/ml up to an MIC of 16 micrograms/ml were analyzed for their penicillin-binding protein (PBP) patterns. With a few exceptions, the great majority of strains with penicillin MICs up to about 0.05 microgram/ml contained the same set of five PBPs with molecular sizes typical of those of susceptible pneumococci. In strains with penicillin MICs of about 0.1 microgram/ml and up, virtually all isolates showed two common features: (i) all isolates showed loss of PBP 1A (98 kilodaltons) with or without a parallel appearance of a "new" PBP that ranged in molecular size between 96 and 97 kilodaltons; and (ii) in strains with penicillin MICs of 0.5 microgram/ml or more, PBP 2B could not be detected on the fluorograms even with very high concentrations of radioactive penicillin. Beyond these two common features, resistant strains with similar penicillin MICs showed a surprising variety of PBP profiles (i.e., in the number and molecular sizes of PBPs), each characteristic of a given isolate. We suggest that in pneumococci remodeling of critical PBPs in more than one way may result in comparable levels of penicillin resistance.
Effect of agitation and terminal subcultures on yield and speed of detection of the Oxoid Signal blood culture system versus the BACTEC radiometric system
Authors: Weinstein M P , Mirrett S , Reimer L G , Reller L B
Published: January 2020

In an initial evaluation, we found the Oxoid Signal blood culture system inferior to the BACTEC radiometric system for detection of some microorganisms causing septicemia (M. P. Weinstein, S. Mirrett, and L. B. Reller, J. Clin. Microbiol. 26:962-964, 1988). To determine whether modified processing of the Oxoid Signal blood culture system could improve its yield and speed of detecting positive cultures relative to the BACTEC radiometric system, we agitated all Oxoid bottles during the first 24 to 48 h of incubation and performed aerobic and anaerobic subcultures of all Oxoid bottles negative after 7 days of incubation. These modifications improved the overall performance of the Oxoid system, particularly with regard to the yield of streptococci, members of the family Enterobacteriaceae, and Haemophilus, Neisseria, and Acinetobacter spp. The speed of detecting positive cultures also was improved, especially within the first 24 h of incubation. However, the BACTEC system still detected more positive cultures (P less than 0.005), especially of obligate aerobes such as Pseudomonas aeruginosa (P less than 0.05) and yeasts (P less than 0.005). The BACTEC system also detected positive cultures earlier than the Oxoid system (e.g., at 24 h of incubation, 70.5% of BACTEC positive cultures detected versus 62.1% of Oxoid positive cultures detected). Further modifications of the Oxoid system which might include a revised medium, additional processing modifications, altered headspace atmosphere, or a complementary second broth medium should be considered, since the system is attractive in concept and is easy to use in the clinical laboratory.
Evaluation of a monoclonal antibody pool for rapid diagnosis of respiratory viral infections
Authors: Stout C , Murphy M D , Lawrence S , Julian S
Published: January 2020

A pool of monoclonal antibodies (MAbP) was evaluated both as a method of cell culture confirmation and as a rapid diagnostic screen for viral infection in respiratory secretions. The MAbP was used in a two-step fluorescent staining procedure on cells harvested from cultures (phase 1) and on exfoliated nasopharyngeal or tracheal cells (phase 2). Antibodies in the MAbP were directed against respiratory syncytial virus, adenoviruses, parainfluenza virus types 1, 2, and 3, and influenza viruses A and B. Individual antiviral antibody stains were used to identify specific viruses from MAbP-positive specimens. In phase 1 (cell culture confirmation only), 241 respiratory specimens were tested. MAbP sensitivity and specificity were 91 and 94%, respectively. In phase 2, 376 respiratory specimens were evaluated by direct staining of exfoliated cells, and these results were compared with results of cell culture isolation. The sensitivity and specificity of the MAbP used in direct specimen testing were 69 and 97%, respectively. These results indicate that the MAbP is highly specific and fairly sensitive for detection of seven different respiratory viruses in one testing procedure. The MAbP is a rapid screening technique for respiratory secretions and is potentially a cost-effective approach to viral detection.
Diagnosis of influenza A virus infections by detection of specific immunoglobulins M, A, and G in serum
Authors: Vikerfors T , Lindegren G , Grandien M , van der Logt J
Published: January 2020

The value of immunoglobulin M (IgM) detection in the early diagnosis of influenza A was examined in a prospective study during an outbreak of influenza A/Philippines/2/82 (H3N2) virus infection in February and March 1986. The IgM response was investigated in sera from 64 adults with influenzalike symptoms; we found a fourfold rise in titer or high titers (greater than or equal to 80) of influenza A virus antibodies by the complement fixation test. The IgM response, analyzed by the hemadsorption immunosorbent technique, was compared with the IgG and IgA responses analyzed by an indirect enzyme-linked immunosorbent assay and the hemadsorption immunosorbent technique, respectively. Antigen detection in nasopharyngeal secretions by immunofluorescence was performed for all patients on admission to hospital. Specific IgM was detected in 86% (55 of 64) of the patients with influenza A. In sera from 36% (18 of 64) of the patients it was detected already on admission. Influenza A virus antigen was detected in nasopharyngeal cells by immunofluorescence on admission in 53% (34 of 64) of the patients. A combination of immunofluorescence and IgM results gave a significantly higher diagnostic rate, 69% (P less than 0.01), on admission than did each of the two tests separately. An IgA serum antibody response was seen in 76% (48 of 64) of the cases but did not contribute to any increase in the diagnostic rate. IgM detection by the hemadsorption immunosorbent technique was found to be a valuable supplement for the diagnosis of influenza A in an early phase of the disease.
Development and evaluation of immunoassay for detection of antibodies to the feline T-lymphotropic lentivirus (feline immunodeficiency virus)
Authors: O'Connor T P , Tanguay S , Steinman R , Smith R , Barr M C , Yamamoto J K , Pedersen N C , Andersen P R , Tonelli Q J
Published: January 2020

The feline T-cell lymphotropic lentivirus (feline immunodeficiency virus) is a recently described feline-specific retrovirus that can produce chronic immunodeficiency-like disorders in cats. A microdilution plate format enzyme-linked immunosorbent assay has been developed to detect the presence of antibody to the virus in feline serum or plasma. Temporal studies performed with experimentally infected animals show that seroconversion can be demonstrated 3 to 4 weeks after exposure to the virus. Results of a serosurvey (n = 1,556 samples) indicate that infection is fairly common in both clinic (5.2%) and sick cat (15.2%) populations. Western blot (immunoblot) and sodium dodecyl sulfate radioimmunoprecipitation assays were developed to confirm microdilution plate test results and to identify peptides specific for the feline immunodeficiency virus. All microdilution plate test positive results and selected negative results were confirmed by one or both of these procedures. These data demonstrate that this microassay plate enzyme-linked immunosorbent assay is a very sensitive and specific test for detection of antibody to the feline immunodeficiency virus.
In vitro differentiation of human-derived Pneumocystis carinii
Authors: Blumenfeld W , Griffiss J M
Published: January 2020

Two major impediments to the development of an in vitro cultivation system for Pneumocystis carinii are lack of an accurate means of quantitation and difficulty in determination of viability over time. Human-derived P. carinii exists as aggregates of cysts and trophozoites in bronchoalveolar lavage fluid. These aggregates persisted over time in an in vitro system consisting of a monolayer of radiated or nonradiated A549 cells, RPMI 1640, and 10% fetal calf serum. Parallel measurements at specified times after introduction into the in vitro system of the number of aggregates, total aggregate area, and total number of cysts varied and appeared to be a function of the number of aggregates initially added into the system. However, cyst density, the number of cysts per unit aggregate area, was independent of the total number of aggregates added into the system. Cyst density was determined by staining an aggregate with a cyst-specific stain, such as toluidine blue, and counterstaining with Diff-Quik, allowing simultaneous visualization of cysts and aggregate area. Preliminary experiments suggested an increase in cyst density over time. Cyst density may be a means of accurate in vitro quantitation of P. carinii.
Comparative field evaluation of the fluorescent-antibody test, virus isolation from tissue culture, and enzyme immunodiagnosis for rapid laboratory diagnosis of rabies
Authors: Bourhy H , Rollin P E , Vincent J , Sureau P
Published: January 2020

The rabies tissue culture infection test (RTCIT) and rapid rabies enzyme immunodiagnosis (RREID) were compared to the fluorescent-antibody test (FAT) with field specimens. At the French National Reference Center for Rabies, 15,248 specimens were analyzed by FAT and RTCIT, and 2,290 of those specimens were also tested by RREID; 818 other specimens were tested by FAT and RREID in 12 laboratories located in Africa, Asia, and Latin America. The sensitivities and specificities of RREID and RTCIT were comparable. This study showed that both tests can be used as backup procedures to confirm FAT. RREID is also strongly recommended for epidemiological studies and for laboratories which are not equipped for performing FAT.
Improved colorimetric assay for detecting influenza B virus neutralizing antibody responses to vaccination and infection
Authors: Tannock G A , Paul J A , Herd R , Barry R D , Reid A L , Hensley M J , Gillett R S , Gillett S M , Lawrance P , Henry R L
Published: January 2020

An automated neutralization test for influenza B virus is described in which antibody titers are determined according to the release of neutral red from infected or uninfected cells of the Madin-Darby canine kidney line. Endpoints are determined in a standard enzyme-linked immunosorbent assay reader. The test requires no expensive immunologic reagents and was used to evaluate responses to both vaccination and natural infection against influenza B virus. Overall responses to vaccination were comparable with those obtained by hemagglutination inhibition, using Tween-ether-split influenza B/Ann Arbor/1/86 virus as the antigen (the HI-TE test). The sensitivities of neutralization responses compared with those obtained by the HI-TE test for two vaccines were 88 and 89%; the specificities were lower at 61 and 60%, respectively. Responses to vaccination, measured by hemagglutination inhibition, were significantly higher with split virus compared with whole virus. However, seroconversion by both the HI-TE and neutralization tests was observed in 5 of 10 individuals from whom virus was detected by either culture of nasal or throat washings or the presence of antigen from immunofluorescence in cells from nasal washings.
Direct identification of serotypes of natural human rotavirus isolates by hybridization using cDNA probes derived from segment 9 of the rotavirus genome
Authors: Zheng B J , Lam W P , Yan Y K , Lo S K , Lung M L , Ng M H
Published: January 2020

Under stringent hybridization conditions, cDNA of segment 9 of the rotavirus genome, which codes for the viral protein VP7, permitted differentiation of serotypes of culture-grown rotaviruses and natural isolates of human rotaviruses directly from clinical specimens. This was evident for the following reasons. (i) The cDNA of one serotype selectively hybridized with the RNA of the same serotype of culture-grown rotaviruses. (ii) Natural isolates of the virus thus identified as serotype 2 were also those which gave the short electrophoretic pattern characteristic of the genome of serotype 2, subgroup 1 viruses. Isolates having the long electrophoretic pattern characteristic of the genome of subgroup 2 viruses were identified as serotype 1, 3, or 4 by this method. (iii) For patients who had previously undergone serological analysis, the serotypes being excreted were the same serotypes against which there was the most marked serum neutralizing antibody response. Although the virus population was genetically diverse, the preponderance of the population prevalent in Guangzhou and Foshan in each of the three successive years between 1982 and 1985 comprised a single serotype. The dominant serotype changed from one year to another, but there was minimal cocirculation of different serotypes in this community. All virus isolates belonged to serotype 1,2,3, or 4, and there was no evidence to suggest that the other human rotavirus serotypes were prevalent in this community.
Use of high-speed rolling to detect respiratory syncytial virus in cell culture
Authors: Sturgill M A , Hughes J H
Published: January 2020

We examined the effect of motion on respiratory syncytial virus (RSV) growth in cell culture. Infected cultures were incubated stationary, rolling, or on an orbital shaker. Enzyme immunoassay (EIA) results for cultures infected with high concentrations of a laboratory strain of RSV were similar for all incubation conditions. However, cultures infected with low concentrations of virus and rolled at 96 rpm had a significantly greater mean EIA optical density (1.78 +/- 0.22) than cultures rolled at 2 rpm (1.42 +/- 0.08) (P less than 0.05). The mean EIA optical density of high-speed-rolled cultures was also significantly greater than for cultures on an orbital shaker (1.25 +/- 0.08) or for stationary cultures (0.21 +/- 0.17) (P less than 0.01). The amount of virus measured by EIA from cultures infected with clinical specimens was also found to be significantly greater at 96 rpm than for stationary cultures. Cultures infected with cell culture isolates were detected significantly earlier at 96 rpm than when stationary. We suggest that high-speed-rolling can be used to enhance the detection of RSV in clinical specimens, especially if the virus is present in low concentrations.
Experience with an enzyme immunoassay for serotyping human group A rotaviruses
Authors: Unicomb L E , Coulson B S , Bishop R F
Published: January 2020

An enzyme immunoassay utilizing neutralizing monoclonal antibodies to VP7 of four human group A rotavirus serotypes successfully typed rotaviruses in 71.4% (568 of 796) of fecal specimens. Sensitivity was enhanced by using homologous capture and detector antibodies. Serotyping was most successful with specimens stored for less than 3 years and containing 10(4) or more particles per ml.
Serum immunoglobulin G antibody subclass response to respiratory syncytial virus F and G glycoproteins after first, second, and third infections
Authors: Wagner D K , Muelenaer P , Henderson F W , Snyder M H , Reimer C B , Walsh E E , Anderson L J , Nelson D L , Murphy B R
Published: January 2020

Serum samples from 31 children who experienced two or three infections with respiratory syncytial virus (RSV) in the first four years of life were tested in an enzyme-linked immunosorbent assay to examine the immunoglobulin G (IgG) subclass responses to the RSV F and G surface glycoproteins associated with primary infection and reinfection. We sought to determine whether the greater degree of glycosylation of the G glycoprotein was reflected in an IgG subclass immune response more like that to a polysaccharide antigen than to a protein antigen. We found that the IgG1/IgG2 ratio of postinfection antibody titers to F was fourfold higher than that to the G glycoprotein after RSV infections 1, 2, and 3. The IgG2 response to the heavily glycosylated G glycoprotein differed from that to a polysaccharide antigen in that the IgG1/IgG2 ratio remained constant with age, whereas the response to a polysaccharide antigen decreased as the IgG2 response increased with age. We also noted that antibody responses to both surface glycoproteins in the IgG1 and IgG2 subclasses reached their maximum levels after RSV infection 2.
Characterization of Borrelia burgdorferi isolates by restriction endonuclease analysis and DNA hybridization
Authors: LeFebvre R B , Perng G C , Johnson R C
Published: January 2020

Genomes of several Borrelia burgdorferi isolates from North America and Europe were characterized by restriction endonuclease analysis and DNA hybridization using labeled B. burgdorferi whole-cell DNA (strain ATCC 35210). Several different restriction and homology patterns were observed among these isolates, indicating genotypic heterogeneity within this genus and species. It was concluded from this study that restriction endonuclease analysis of B. burgdorferi whole-cell DNA may be a reliable and accurate method for identifying strains or genotypes of the Lyme disease agent.
Evaluation of DNA colony hybridization and other techniques for detection of virulence in Yersinia species
Authors: Robins-Browne R M , Miliotis M D , Cianciosi S , Miller V L , Falkow S , Morris J G
Published: January 2020

The virulence of yersiniae varies according to (i) species and biotype and (ii) possession of a 67- to 72-kilobase virulence plasmid. Y. pestis, Y. pseudotuberculosis, and biotypes 1B, 2, 3, 4, and 5 of Y. enterocolitica are inherently virulent but express full virulence only when in possession of a virulence plasmid. Other Yersinia species and biotypes 1A and 3B of Y. enterocolitica are seldom implicated in disease. In this study, we prepared DNA probes from eight nonoverlapping regions of the virulence plasmid of a strain of Y. enterocolitica and from the inv and ail chromosomal loci responsible for the invasive capacity of Y. enterocolitica and Y. pseudotuberculosis. The probes were used in colony hybridization experiments to investigate 156 yersiniae of various species and biotypes and of differing virulence. Probes prepared from the inv gene of Y. pseudotuberculosis hybridized with Y. pseudotuberculosis and Y. pestis only, whereas an analogous probe prepared from Y. enterocolitica hybridized with all species and biotypes of yersiniae (but not with other bacteria) regardless of virulence or potential virulence. Probes prepared from the ail region of Y. enterocolitica reacted almost exclusively with Y. enterocolitica strains of pathogenic biotypes. Probes prepared from the virulence plasmid of a serogroup O:8, biotype 1B isolate of Y. enterocolitica identified virulent yersiniae in all species with a high degree of sensitivity and specificity. These probes did not react with yersiniae of avirulent biotypes or species. Of the other assays of virulence evaluated (calcium dependence, binding of crystal violet, and pyrazinamidase activity), binding of crystal violet provided a simple means for identifying plasmid-bearing strains.
Seroepidemiology of human herpesvirus 6 infection in normal children and adults
Authors: Okuno T , Takahashi K , Balachandra K , Shiraki K , Yamanishi K , Takahashi M , Baba K
Published: January 2020

Sera from normal subjects were examined for reactivity to human herpesvirus 6 (HHV-6) by the anticomplement immunofluorescence test. Of a total of 179 serum specimens from donors aged from under 10 to 59 years, 141 specimens showed positive reactivity against HHV-6. The positive rate was 70 to 83% for all age groups, and there were no substantial differences in the positive rates. Sera from younger children aged from 0 to 21 months were then examined in detail. The antibody-positive rate of children aged from 0 to 5 months decreased from 52 to 5%, but it gradually increased by 12 months. Almost all children had the antibody against this virus after 13 months of age.
Clostridium perfringens food poisoning: use of serotyping in an outbreak setting
Authors: Gross T P , Kamara L B , Hatheway C L , Powers P , Libonati J P , Harmon S M , Israel E
Published: January 2020

An outbreak of Clostridium perfringens food poisoning occurred among attendees of a firehouse luncheon. The predominant symptoms of diarrhea (100%) and abdominal pain (81%) among case-patients, the mean incubation period (13.4 h), and the mean duration of illness (21.2 h) were all characteristic of C. perfringens enteritis. Roast beef, although not epidemiologically implicated, was the most likely vehicle of transmission. Fecal specimens from case-patients contained a median C. perfringens spore count of greater than 10(6) and yielded isolates that were heat sensitive and predominantly nonhemolytic, produced C. perfringens enterotoxin A, and, in the majority of specimens (four of five), were identical in serotype. Food samples were negative. This outbreak demonstrates that following enumeration of C. perfringens from a suitable number of fecal specimens from case-patients, serotyping of the isolates may be helpful in implicating C. perfringens as the cause of foodborne illness. This is especially true when implicated food items test negative or are no longer available for testing.
Comparison of the Pasteurella haemolytica A1 envelope proteins obtained by two cell disruption methods
Authors: Simons K R , Morton R J , Mosier D A , Fulton R W , Confer A W
Published: January 2020

The French pressure cell and sonication methods of bacterial cell disruption were compared for the evaluation of surface proteins from Pasteurella haemolytica A1. Several protein bands were quantitatively different when compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and densitometry. With densitometry, sonicated preparations had higher concentrations of the 102-, 83-, 50-, 38-, and 30-kilodalton (kDa) proteins; French pressure cell preparations had higher concentrations of the 96-, 65-, and 42-kDa proteins. Qualitative differences between these two disruption methods were evident at the 102-, 96-, 91-, 50-, 38-, and 30-kDa protein bands. However, significant differences (P less than 0.05) were detected between the two methods at only the 102-, 96-, 91-, and 50-kDa bands.
Direct isolation of atypical thermophilic Campylobacter species from human feces on selective agar medium
Authors: Walmsley S L , Karmali M A
Published: January 2020

Campylobacter upsaliensis is the name which has been proposed for a new group of thermophilic campylobacter strains which differ from C. jejuni and C. coli in having a negative or weak catalase reaction. Primary isolation of these strains from human feces has been achieved only by use of filtration techniques. We report here direct isolation of strains corresponding to C. upsaliensis from stools of six children. The strains were isolated on a newly described campylobacter-selective medium. The strains were oxidase positive, hippurate negative, nitrate positive, negative for H2S in triple sugar iron, and susceptible to cephalothin (30-micrograms disk) and nalidixic acid (30-micrograms disk), and they grew at 37 and 43 degrees C, but not at 25 degrees C. The selective medium used was a blood-free, charcoal-based medium consisting of Columbia agar base, activated charcoal, cefoperazone (32 micrograms/ml), vancomycin (20 micrograms/ml), and cycloheximide (100 micrograms/ml). The medium supported the growth of the weakly reacting or catalase-negative strains, with colony counts equivalent to those obtained on antibiotic-free horse blood agar. These strains could not be isolated directly from stool on Skirrow medium, and colony counts confirmed that this medium could not support a low inoculum of these organisms. The clinical significance of these strains is unknown. We conclude that C. upsaliensis can be isolated directly from stool by using a selective medium, without the need for filtration.
Purification, characterization, and immunological cross-reactivity of alginates produced by mucoid Pseudomonas aeruginosa from patients with cystic fibrosis
Authors: Pedersen S S , Espersen F , Høiby N , Shand G H
Published: January 2020

Alginates from nine mucoid Pseudomonas aeruginosa isolates from patients with cystic fibrosis were purified by repeated ethanol precipitation, nuclease digestion, anion-exchange chromatography, dialysis, and lyophilization. Uronic acid constituted 72% of the dry weight when mannuronolactone was used as the internal standard in the carbazole-borate assay for uronic acids. The average degree of acetylation was 16%, and the ratio of mannuronic acid to gluluronic acid was 4.7. No homopolymeric blocks of guluronic acid were found when analyzed by nuclear magnetic resonance spectroscopy. Contaminating proteins were denatured by heating, and during purification the content of protein relative to alginate fell from 566 to 0.9%. The content of lipopolysaccharide was 0.012%. No immunological or biological activity was attributable to the protein or lipopolysaccharide content as estimated by immunoblotting, enzyme-linked immunosorbent assay (ELISA), and a neutrophil chemotaxis assay. Rabbits were hyperimmunized with P. aeruginosa alginates and alginate from the seaweed Laminaria hyperborea, and an ELISA that detected alginate-specific antibodies was developed. Antibodies to P. aeruginosa alginate were detected by ELISA in 1:4,000 dilutions of serum from patients with cystic fibrosis with chronic P. aeruginosa lung infection. The serological cross-reactions between serum from the nine patients with cystic fibrosis and the corresponding P. aeruginosa alginates were investigated and showed considerable heterogeneity. This finding indicates that P. aeruginosa alginate from more than one P. aeruginosa strain should be used in serological tests. There was no serological cross-reactivity between P. aeruginosa and Laminaria hyperborea alginate in either rabbits or patients with cystic fibrosis.
Diagnosis of herpes simplex virus infection in a clinical setting by a direct antigen detection enzyme immunoassay kit
Authors: Dascal A , Chan-Thim J , Morahan M , Portnoy J , Mendelson J
Published: January 2020

A commercial 4-h direct herpes simplex virus (HSV) antigen detection enzyme immunoassay (EIA) kit (Du Pont Herpchek) was evaluated by using 273 clinical specimens obtained in a hospital-based infectious disease practice. The EIA was compared with a standard culture method in which WI38 cells were inoculated within 20 min of sample collection. Cultures were observed for 2 weeks, and positive findings were confirmed by fluorescein-labeled monoclonal antibody (FA) staining. The values for the overall HSV detection rate were 40.7% by the standard culture method and 41.4% by EIA. In eight cases, the EIA was positive, while the culture method was negative; however, clinical data and confirmatory blocking EIA suggested that a true HSV infection was present. For six FA-confirmed, culture-positive samples, the direct EIA was negative; however, an EIA performed on the supernatants of these cultures was positive, suggesting that the failure of the EIA to detect these samples was not due to lack of strain specificity of the test. After confirmatory tests of standard culture and EIA discrepant results, the overall sensitivity of the test was 95.0% (113 of 119) and the specificity was 100% (154 of 154).
Diagnosis of Mycobacterium bacteremia in patients with acquired immunodeficiency syndrome by direct examination of blood films
Authors: Eng R H , Bishburg E , Smith S M , Mangia A
Published: January 2020

Thirty acquired immunodeficiency syndrome patients with mycobacterial bacteremia documented by Du Pont Isolator (Du Pont Co., Wilmington, Del.) blood cultures underwent microscopic examination of buffy coat blood smears. Of 30 patients, 14 were culture positive for Mycobacterium avium-Mycobacterium intracellulare complex and 1 was positive for M. tuberculosis. Of 15 culture-positive patients, 13 had identifiable organisms on Kinyoun- or auramine-stained direct blood smears.
Comparison of a new rapid test (TestPack Rotavirus) with standard enzyme immunoassay and electron microscopy for the detection of rotavirus in symptomatic hospitalized children
Authors: Brooks R G , Brown L , Franklin R B
Published: January 2020

We compared a new, rapid, qualitative test for rotavirus (TestPack Rotavirus; Abbott Laboratories, North Chicago, Ill.) with another enzyme immunoassay (Pathfinder Rotavirus; Kallestad Laboratories, Inc., Austin, Tex.) and electron microscopy to determine its clinical utility in a population of symptomatic hospitalized children. In the first part of the study, 100 frozen stool samples were tested. The results after resolution with a blocking reagent showed a sensitivity of only 50% and a specificity of 88% for TestPack Rotavirus. In the second part of the study, we tested TestPack Rotavirus on 100 fresh, unfrozen samples. The results (sensitivity/specificity) were as follows: TestPack Rotavirus, 95/90%; Pathfinder Rotavirus, 84/98%; direct electron microscopy, 63/100%. Although it was not as sensitive or specific as immune electron microscopy, TestPack Rotavirus was more sensitive than direct electron microscopy or Kallestad Pathfinder Rotavirus. TestPack Rotavirus represents a rapid, qualitative method for the detection of rotavirus in stools of symptomatic children.
Temporal variation of Malaysian rotavirus electropherotypes
Authors: Rasool N , Othman R Y , Adenan M I , Hamzah M
Published: January 2020

An analysis of rotavirus electropherotypes circulating in Kuala Lumpur, Malaysia, over 7 years showed that all except 1 of the 360 electropherotypes encountered were characteristic of group A rotaviruses. The long electropherotype predominated annually, and there was a rarity of short electropherotypes. Extensive genome variability and cocirculation of different electropherotypes were observed annually. A sequential appearance of the predominant electropherotype was observed in all years of the study, except for 1985 and 1988, when one electropherotype predominated throughout the study periods. There was no shift in the predominant electropherotype over a 6-year period.
Antigenic characterization of swine rotaviruses in Argentina
Authors: Mattion N M , Bellinzoni R C , Blackhall J O , La Torre J L , Scodeller E A
Published: January 2020

Fecal samples from 156 diarrheic piglets were collected from several herds located in two main breeding areas of Argentina. Rotaviruses were detected in 60 samples (38.4%) by polyacrylamide gel electrophoresis and in 55 samples by a group A-specific enzyme-linked immunosorbent assay (ELISA). All samples which were positive by polyacrylamide gel electrophoresis and negative by ELISA had elicited atypical electropherotypes resembling those of group B or C. ELISA-positive samples showing genome rearrangements were also detected (R.C. Bellinzoni, N.M. Mattion, O.R. Burrone, S.A. González, J.L. La Torre, and E.A. Scodeller, J. Clin. Microbiol. 25:952-954, 1987; N.M. Mattion, S.A. González, O.R. Burrone, R.C. Bellinzoni, J.L. La Torre, and E.A. Scodeller, J. Gen. Virol. 69:695-698, 1988). By subgrouping with monoclonal antibodies, it was found that of 32 positive samples, 13 belonged to subgroup I, 2 belonged to subgroup II, 2 samples had both specificities, and 15 samples were neither subgroup I nor subgroup II (non-I/II). In addition, 10 samples were adapted to grow in tissue culture, cloned, and serotyped by means of neutralization assays. Two samples were classified as serotype 5, and none of them were classified as serotype 4. The other strains showed only a one-way relationship with serotype 5 and can be tentatively classified as new porcine serotypes. Two samples with rearranged genomes had a one-way relationship with antiserum to human strain 69M, which displays a supershort electropherotype and was classified as a new human serotype (S. Matsuno, A. Hasegawa, A. Mukoyama, and S. Inouye, J. Virol. 54:623-624, 1985). At one farm, similar rearranged strains were detected during three successive years. Serotype changes were found between the isolates of the first and the second year, suggesting that a high degree of antigenic variability went on during continuous circulation of these strains in the field.
Evaluation of the Minitek Gram-Positive Set for identification of streptococci isolated from bovine mammary glands
Authors: Watts J L
Published: January 2020

A total of 127 isolates were used to evaluate the Minitek Gram-Positive Set for identification of streptococci cultured from bovine mammary glands. The overall accuracy of the Minitek Gram-Positive Set was 34.6%. Of 12 Streptococcus agalactiae strains, 4 (33.3%) were correctly identified. Of 43 Streptococcus dysgalactiae strains, 32 (74.4%) were identified correctly. Of 44 Streptococcus uberis strains, 42 (95.5%) were identified as Enterococcus spp. Poor performance was attributed to the limited number of veterinary strains in the data base. Incorporation of large numbers of veterinary isolates into the data base is needed for further development of this system.
Rapid purification of staphylococcal enterotoxin B by high-pressure liquid chromatography
Authors: Strickler M P , Neill R J , Stone M J , Hunt R E , Brinkley W , Gemski P
Published: January 2020

The Staphylococcus aureus enterotoxins represent a group of proteins that cause emesis and diarrhea in humans and other primates. We have developed a rapid two-step high-pressure liquid chromatography (HPLC) procedure for purification of staphylococcal enterotoxin B (SEB). Sterile filtrates (2.5 liters) of strain 10-275 were adsorbed directly onto a reversed-phase column (50 mm by 30 cm Delta Pak; 300 A [30 nm], 15 microns, C18). SEB was obtained by using a unique sequential gradient system. First, an aqueous ammonium acetate to acetonitrile gradient followed by an aqueous trifluoroacetic acid (TFA) wash was used to remove contaminants. A subsequent TFA to acetonitrile-TFA gradient eluted the bound SEB. Further purification was obtained by rechromatography on a cation-exchange column. From 35 to 45% of the SEB in starting filtrates was recovered. Analysis by immunoblotting of samples separated on sodium dodecyl sulfate-polyacrylamide gels indicated that HPLC-purified SEB exhibited immunological and biochemical properties similar to those of the SEB standard. Induction of an emetic response in rhesus monkeys showed that the HPLC-purified toxin also retained biological activity.
Molecular species of R-protein antigens produced by clinical isolates of group B streptococci
Authors: Flores A E , Ferrieri P
Published: January 2020

Clinical isolates of group B streptococci from body fluids and mucosal surfaces were examined for production of a trypsin-resistant antigen known as R protein. R protein was extracted with 1% trypsin from cells grown in a semidefined medium. The extracts were tested by immunodiffusion in agarose with a panel of antisera for detection and precise identification of the four species of R protein described by Wilkinson. R antigen was present in 49 of 131 (37%) of the strains tested. Analysis by serotype revealed that 0 of 2 type Ia, 0 of 11 Ib, 1 of 16 (6%) Ia/c, 12 of 15 (80%) II, 0 of 20 II/c, 35 of 49 (71%) III, 0 of 6 IV, and 1 of 12 (8%) nontypeable strains produced R antigen. Production of the R protein and the trypsin-resistant or alpha component of the c protein appeared to be mutually exclusive. R antigen was more prevalent in isolates from blood (50%) than in those from mucosal sites (27%) for type II strains; no difference was seen for type III strains from these sites. Concordant results were obtained with five paired body fluid-mucosal surface isolates from individual patients and with isolates from 17 mother-baby pairs. The most frequent species of R antigen was R4 (45 of 49), followed by R1 (4 of 49). These two species of R protein were biochemically (trypsin resistant and pepsin sensitive) and immunologically identical to the R-protein antigens produced by prototype strains of groups A, B, and C streptococci.
Activities and sources of beta-lactamase in sputum from patients with bronchiectasis
Authors: Dragicevic P , Hill S L , Burnett D , Merrikin D , Stockley R A
Published: January 2020

beta-Lactamase activity was measured in secretions from patients with bronchiectasis. Of 28 sputum samples, 23 contained measurable amounts of activity; values were significantly higher (P less than 0.01) in purulent samples than in mucoid or mucopurulent samples. beta-Lactamase activity was usually present in saliva collected before and between sputum expectorations, although values for sputum were higher than for either group of saliva samples (P less than 0.025 and P less than 0.005, respectively). This difference suggests that at least part of sputum beta-lactamase activity originates in the bronchial tree. Detailed microbiological study of a further eight specimens (seven were beta-lactamase positive) led to the isolation of Haemophilus influenzae from six, although only two of these isolates were beta-lactamase positive. Several other beta-lactamase-producing organisms were also isolated, including Staphylococcus aureus (n = 3), Escherichia coli (n = 1), Proteus spp. (n = 1), and Bacteroides spp. (n = 3). Size-exclusion high-performance liquid chromatography of the sputum showed several peaks of beta-lactamase activity which usually coeluted in fractions similar to those of their beta-lactamase-positive isolates. Therefore, sources of sputum beta-lactamases are often bacteria not considered truly pathogenic or not isolated during routine bacteriological assessment. These observations should be considered when embarking on antimicrobial therapy in bronchiectatic patients and suggest that increased dosages of penicillins are indicated.
Semisolid blood-free selective-motility medium for the isolation of campylobacters from stool specimens
Authors: Goossens H , Vlaes L , Galand I , Van den Borre C , Butzler J P
Published: January 2020

Isolation of Campylobacter jejuni and C. coli from stool specimens is done by growing campylobacter colonies on solid selective media with or without blood. However, recognition of these colonies can be difficult. Therefore, we decided to evaluate an isolation procedure based on the swarming of campylobacters through a semisolid medium. We developed a semisolid blood-free selective motility (SSM) medium which is composed of Mueller-Hinton broth with 0.4% agar and supplemented with cefoperazone (30 micrograms/ml) and trimethoprim (50 micrograms/ml). The SSM medium was compared with our previously described Butzler Medium Virion (Goossens et al., J. Clin. Microbiol. 24:840-843, 1986) and blood-free medium (Bolton and Coates, J. Appl. Bacteriol. 54:115-125, 1983) with cefoperazone (32 micrograms/ml) (Bolton et al., J. Clin. Pathol. 37:956-957, 1986). Of 1,890 routine stool specimens tested, 100 were found to be positive for campylobacters: 95 were recovered with the SSM medium, 94 with the Virion medium, and 90 with the blood-free medium. The SSM medium performed equally well whether it was incubated in the special incubator or the candle jar. Only 4.4 and 7.3% of the plates grew contaminating fecal flora when incubated in the special incubator and the candle jar, respectively. Clearly the SSM medium is easy, quick, cheap, sensitive, and more selective than any other medium which has been developed so far and does not require the addition of blood. We believe that this medium has a future in the routine microbiology laboratory in developed as well as in developing countries.
Effects of amino acids on colony phenotype of Neisseria gonorrhoeae
Authors: Saikh K U , Mitra S , Bhattacharyya F K
Published: January 2020

The effects of amino acid supplementation of commercially available semidefined GC media (Oxoid GC medium and Difco GC base) on the stability of the virulent colony phenotype of Neisseria gonorrhoeae were studied. The -SH-containing amino acids, together with glycine, were found to cause segregation of avirulent colony types followed by growth inhibition after addition of 40 mg/100 ml. Proline and arginine were found to change the colony phenotype profile significantly without reducing the viable count. Arginine supplementation in both media caused a progressive loss of virulent colony types, whereas proline had the opposite effect.
Comparison of four charcoal media for the isolation of Bordetella pertussis
Authors: Hoppe J E , Schwaderer J
Published: January 2020

Charcoal-horse blood agar with 40 micrograms of cephalexin per ml, charcoal-horse blood agar with 3 micrograms of lincomycin per ml, charcoal agar with 3 micrograms of lincomycin per ml, and Legionella (buffered charcoal-yeast extract) agar with 3 micrograms of lincomycin per ml were compared for isolation of Bordetella pertussis. Charcoal-horse blood agar with 40 micrograms of cephalexin per ml gave the best results, with a B. pertussis recovery rate of 100%. Growth was most rapid and the mean number of colonies was highest on this agar, and growth of pharyngeal flora was completely suppressed.
Repeat antimicrobial susceptibility testing of identical isolates
Authors: Thomson R B , File T M , Burgoon R A
Published: January 2020

Duplicate antimicrobial susceptibility test results were reviewed over a 1-year period to determine whether repeat testing of sequential isolates with the same identification from the same patient and specimen site was necessary. In our institution, repeat testing is always needed for coagulase-negative staphylococci and Pseudomonas aeruginosa and is needed after 3 days for members of the family Enterobacteriaceae, but it is not routinely necessary for Staphylococcus aureus.
Isolation and characterization of monoclonal antibodies against alkaline phosphatase of Pseudomonas aeruginosa
Authors: Husson M O , Mielcarek C , Gavini F , Caron C , Izard D , Leclerc H
Published: January 2020

Monoclonal antibodies against the alkaline phosphatase of Pseudomonas aeruginosa were produced from spleen cells of BALB/c mice primed with purified alkaline phosphatase of P. aeruginosa ATCC 10145 and SP20/Ag-14 myeloma cells. The eight stable clones established produced antibodies that reacted by enzyme-linked immunosorbent and indirect immunofluorescence assays with all bacterial strains of P. aeruginosa, including the 17 serotypes and two nontypable strains. Three of the clones cross-reacted only with some Pseudomonas species of the rRNA homology group I defined by N. J. Palleroni (in N. R. Krieg and J. G. Holt, ed., Bergey's Manual of Systematic Bacteriology, 8th ed., p. 140-218, 1984). The other clones also interacted with other species, including Pseudomonas acidovorans and Xanthomonas maltophilia. Because other species of the genera Aeromonas and Acinetobacter and species of the family Enterobacteriaceae were not detected by these monoclonal antibodies, the antibodies could be used as reagents for routine detection of P. aeruginosa in clinical specimens. Interactions of the antibodies with other Pseudomonas species such as P. fluorescens and P. stutzeri are not important, since these species are susceptible to the same antipseudomonal agents.
Susceptibility of methicillin-resistant Staphylococcus aureus to lysostaphin
Authors: Huber M M , Huber T W
Published: January 2020

One hundred and eleven isolates of methicillin-resistant Staphylococcus aureus recovered from patients at the Olin E. Teague Veterans Center from March 1983 to April 1987 were as susceptible to lysis by lysostaphin as methicillin-susceptible S. aureus controls were.
Stomatococcus mucilaginosus catheter-associated infection with septicemia
Authors: Poirier L P , Gaudreau C L
Published: January 2020

A case of Stomatococcus mucilaginosus catheter-associated infection is reported. The organism was isolated from the catheter and from two blood cultures.
Comparison of methods for determining DNase and phosphatase activities of staphylococci
Authors: Langlois B E , Harmon R J , Akers K , Aaron D K
Published: January 2020

A greater percentage of DNase-positive strains was detected with DNase test agar than with DNase test agar containing 0.005% methyl green or 0.005% toluidine blue (P less than 0.01). No significant differences were obtained in the percentage of phosphatase-positive strains with the four methods compared. On the basis of ease of use, P agar containing para-nitrophenylphosphate disodium (0.495 mg/ml) would be the preferred method for determining phosphatase activity of staphylococci.
Gardnerella vaginalis bacteremia from pulmonary abscess in a male alcohol abuser
Authors: Legrand J C , Alewaeters A , Leenaerts L , Gilbert P , Labbe M , Glupczynski Y
Published: January 2020

A case of Gardnerella vaginalis bacteremia is reported. This bacteremia occurred in a male alcohol abuser who developed definite signs of pulmonary abscess and empyema. Streptococcus milleri grew from another blood culture, but Gardnerella vaginalis was also isolated from a bronchoscopic aspirate and pleural drainage sample as part of mixed flora containing anaerobes, Streptococcus species, Neisseria sicca, and a Haemophilus sp. We discuss the possible pathogenic character of G. vaginalis outside the genital tract from a review of the literature.
Detection of Cryptosporidium oocysts in human fecal specimens by an indirect immunofluorescence assay with monoclonal antibodies
Authors: Rusnak J , Hadfield T L , Rhodes M M , Gaines J K
Published: January 2020

An indirect fluorescence assay (IFA) with monoclonal antibodies developed for the detection of Cryptosporidium oocysts in fecal smears (Meridian Diagnostics, Inc., Cincinnati, Ohio) was compared with the Zeihl-Neelsen-modified acid-fast stain (MAFS) in 119 human fecal specimens collected between 1984 and 1987. The sensitivity of the IFA was 100%; all 56 specimens positive by MAFS exhibited fluorescence. There were 63 specimens negative for Cryptosporidium sp. by MAFS; of these, 61 were negative by IFA (97% specificity). This discrepancy may reflect an increased sensitivity of the IFA to detect oocysts that were not visualized by MAFS because of faint staining or a paucity of organisms. On average, the IFA required less time than the MAFS (1 versus 5 min, respectively) when only rare or few oocysts were present. Cost comparison of reagents showed the IFA to be three times more expensive. The IFA offers a reasonable alternative to the MAFS because of its high sensitivity and specificity, the simplicity of performing it and interpreting results, and its capability of providing a definitive diagnosis of Cryptosporidium oocysts.
Extensive colonization of the porcine colonic epithelium by a spirochete similar to Treponema innocens
Authors: Jacques M , Girard C , Higgins R , Goyette G
Published: January 2020

Specimens of colonic mucosa from two pigs with diarrhea were examined by light and electron microscopy. The epithelial surfaces of both pigs were extensively colonized by large spirochetes morphologically compatible with Treponema hyodysenteriae or Treponema innocens. The microorganisms were intimately attached end-on to the luminal cells. A weakly beta-hemolytic spirochete similar to T. innocens was isolated from the colon of one of the pigs.