bluuebunny/crossref_metadata_2025 huggingface dataset:
Electrophoretic karyotyping of typical and atypical Candida albicans
Authors: Mahrous M , Lott T J , Meyer S A , Sawant A D , Ahearn D G
Published: January 2020

Electrophoretic karyotypes of atypical isolates of Candida albicans, e.g., strains that were germ tube negative, failed to express proteinase activity, demonstrated low virulence for mice, formed hyperchlamydoconidia, produced hyperhyphae, or were sucrose negative (including the type strain of Candida stellatoidea), were compared with those of typical C. albicans. Karyotypes of whole-cell DNA of classical C. albicans examined with transverse alternating-field electrophoresis under specific conditions were composed of seven DNA bands with a specific migration pattern. Certain atypical strains and representatives of the three serotypes of C. stellatoidea produced discrete karyotypes with 5 to 10 bands. All isolates demonstrated a significant degree of DNA relatedness, suggesting their conspecificity. Densitometric tracings of DNA bands provided an objective and standardized method for comparing bands within the gels.
Comparison of antibody, antigen, and metabolite assays for hospitalized patients with disseminated or peripheral candidiasis
Authors: Bougnoux M E , Hill C , Moissenet D , Feuilhade de Chauvin M , Bonnay M , Vicens-Sprauel I , Pietri F , McNeil M , Kaufman L , Dupouy-Camet J
Published: January 2020

Repeat serum samples from 22 patients with proven disseminated candidiasis and 42 with simple peripheral colonization were assayed for Candida antibodies by coelectrosyneresis, immunoprecipitation, and A and B immunofluorescence, for metabolites by D-arabinitol measurement, and for antigens by the mannan immunoassay and Cand-tec latex agglutination (mean number of samples tested, 2.5 per patient). For the antibody and metabolite assays, the results showed no statistical difference between the two groups. By contrast, the results of both antigen assays were positive for a significantly larger number of patients with disseminated candidiasis than of those with simple peripheral colonization. Results were regardless of whether the patients were neutropenic. They were not predictive of death. We calculated that the mannan antigen assay had 29% sensitivity and 97% specificity for the diagnosis of disseminated candidiasis. Likelihood ratios of a positive and a negative result of this test were 9.2 and 0.7, respectively, for this diagnosis. In the latex agglutination test, likelihood ratios were 2.5, 1.5, 1.6, and 0.3 when the test was positive for dilutions of 1/8, 1/4, and 1/2 and was negative, respectively.
Epidemiologic investigations of Yersinia enterocolitica and related species: sources, frequency, and serogroup distribution
Authors: Bissett M L , Powers C , Abbott S L , Janda J M
Published: January 2020

During an 11-year period (1978 to 1989), over 300 strains of Yersinia spp. (excluding Y. pestis and Y. pseudotuberculosis) were recovered from a variety of gastrointestinal and extraintestinal sites in patients in California. Over the 11-year period, Y. enterocolitica serogroup O:3 predominated, although a shift in the relative frequency of this serogroup was observed during this interval, increasing dramatically during the years 1984 to 1989. Of the remaining Y. enterocolitica isolates, over 40% were identified as belonging to serogroups generally considered to be nonpathogenic, although many of these isolates were recovered in association with milder cases of gastroenteritis. The results suggest a changing and expanding spectrum of Y. enterocolitica serogroups associated with various gastrointestinal and systemic infections.
Three new major somatic antigens of Pseudomonas aeruginosa
Authors: Liu P V , Wang S
Published: January 2020

Three new major somatic antigens of Pseudomonas aeruginosa have been described and added to the 17 existing groups in the International Antigenic Typing System (P. V. Liu, H. Matsumoto, H. Kusama, and T. Bergan, Int. J. Syst. Bacteriol. 33:256-264, 1983). These newly recognized antigens are present in some strains of P. aeruginosa that have been used as type strains of various typing schemata but have not been recognized because of cross-reactions of these strains with antisera of other serogroups with minor antigens. The lists of corresponding serogroups of various serotyping schemata have been revised accordingly.
Use of highly specific DNA probes and the polymerase chain reaction to detect Mycobacterium paratuberculosis in Johne's disease
Authors: Vary P H , Andersen P R , Green E , Hermon-Taylor J , McFadden J J
Published: January 2020

DNA probes that hybridize to a mycobacterial insertion sequence, IS900, present in multiple copies in the genome of Mycobacterium paratuberculosis were found to be highly specific for M. paratuberculosis. DNA sequences derived from IS900 were used to prepare DNA primers for detection and identification of M. paratuberculosis by the polymerase chain reaction. Highly specific direct detection of M. paratuberculosis DNA in feces from cattle with Johne's disease was obtained. The polymerase chain reaction test had a sensitivity equal to or greater than that obtained by standard culture techniques and was much more rapid, taking only hours compared with 6 to 12 weeks for culture.
Evaluation of an rRNA-derived oligonucleotide probe for culture confirmation of Neisseria gonorrhoeae
Authors: Rossau R , Duhamel M , Van Dyck E , Piot P , Van Heuverswyn H
Published: January 2020

The reliability of an rRNA-derived oligonucleotide probe for Neisseria gonorrhoeae was tested with 187 N. gonorrhoeae isolates, 81 Neisseria meningitidis isolates, and several strains of other bacterial species. The probe proved to be 100% specific and 100% sensitive. N. gonorrhoeae cells could also be reliably identified in contaminated cultures with the oligonucleotide probe. The 2.6-megadalton cryptic plasmid used as a probe for N. gonorrhoeae was shown to be less sensitive, detecting 179 of 181 N. gonorrhoeae isolates.
Immunodetection of human T-cell lymphotropic virus type I core protein in biological samples by using a monoclonal antibody immunoassay
Authors: Papsidero L , Swartzwelder F , Sheu M , Montagna R , Ehrlich G , Bhagavati S , Dosik H , Sninsky J , Poiesz B
Published: January 2020

A monoclonal antibody-based enzyme immunoassay (EIA) has been developed for detection of human T-cell lymphotropic virus type I (HTLV-I) core protein. The monoclonal antibody (clone 6.11) specifically recognizes the p19 gag gene-encoded protein of the virus. The EIA was over 100 times more sensitive than reverse transcriptase measurement and was capable of responding to less than 500 pg of whole-virus lysate. The assay exhibited type specificity in that HTLV-II antigens failed to produce a positive signal. In addition, a panel of other viruses demonstrated no antigenic cross-reactivity. These included herpesviruses, measles virus, human immunodeficiency viruses, and others. Viral p19 was followed during the course of density gradient ultracentrifugation in the presence of detergent, where it was noted to associate with viral membrane proteins. In comparison, reverse transcriptase activity localized in fractions of higher density containing envelope-free cores. Of clinical interest, the EIA was used to detect HTLV-I antigen in the viral cultures of patients with HTLV-I-associated myelopathies and from symptom-free individuals with proviral integration.
Antimicrobial susceptibility testing of less commonly isolated Haemophilus species using Haemophilus test medium
Authors: Jorgensen J H , Howell A W , Maher L A
Published: January 2020

Haemophilus test medium (HTM) was developed recently for dilution and disk diffusion antimicrobial agent susceptibility testing of Haemophilus influenzae. The application of HTM to the testing of other, less frequently encountered Haemophilus species recovered from humans was evaluated in this study by using commercially prepared HTM (BBL Microbiology Systems, Cockeysville, Md.) in broth microdilution and agar disk diffusion susceptibility tests with 18 antimicrobial agents. A total of 93.3% of 90 isolates belonging to six Haemophilus species provided acceptable growth in HTM agar disk diffusion tests, while only 63.3% (57 of 90) provided acceptable growth in the broth microdilution tests. However, HTM agar dilution testing provided an alternative method for those strains (primarily H. haemolyticus) which failed to grow adequately in broth. Based on the latest National Committee for Clinical Laboratory Standards guidelines (standard M2-T4) for interpretation of HTM disk tests of H. influenzae, the overall very major, major, and minor errors for all 18 drugs and six species tested were 0.2, 0.7, and 3.4%, respectively. Thus, the use of HTM in agar or broth susceptibility tests can be recommended for testing the less commonly encountered Haemophilus species by using the same test conditions and interpretive guidelines developed for H. influenzae.
Parasite detection efficiencies of five stool concentration systems
Authors: Perry J L , Matthews J S , Miller G R
Published: January 2020

Fresh fecal material that was free of ova and parasites was pooled with 10% Formalin in a 1:4 ratio to prepare a standard specimen. Portions of 100 ml of this specimen were individually seeded with Cryptosporidium oocysts, Entamoeba coli, Entamoeba histolytica, and Giardia lamblia cysts; ova of Necator americanus; and Strongyloides larvae. Appropriate volumes of each parasite suspension were used to evaluate the Fecal Concentrator Kit (Remel, Lenexa, Kans.), Fecal Parasite Concentrator (Evergreen Scientific, Los Angeles, Calif.), Para-Pak Macro-Con (Meridian Diagnostics, Inc., Cincinnati, Ohio), and Trend FeKal CON-Trate (Trend Scientific, Inc., St. Paul, Minn.). A standardized gauze filtration method was used as the reference procedure. Tests were performed in triplicate with each individual parasite-concentrator combination, with three slides examined from each sediment. All of the systems effectively concentrated parasites compared with direct examination of unconcentrated fecal material. The Fecal Concentrator Kit provided the best overall performance. Clarity of sediment, lack of debris, and uniformity of background material were found to be important considerations for microscopic detection of parasites in concentrated specimens.
Alga associated with diarrhea in patients with acquired immunodeficiency syndrome and in travelers
Authors: Long E G , Ebrahimzadeh A , White E H , Swisher B , Callaway C S
Published: January 2020

Spherical bodies resembling coccidian oocysts and measuring 8.0 to 9.0 microns in diameter were seen in the stools of eight persons with explosive, watery diarrhea. Seven had recently traveled to tropical countries, mostly in the Caribbean, and four had acquired immunodeficiency syndrome. The structures were easily discernible in wet mounts by light microscopy and contained variable numbers of granular inclusions, but were refractory to, or stained partially with, 12 commonly used laboratory stains. Electron microscopy revealed an outer fibrillar coat, a thin cell wall, granules, and organelles which were not surrounded by membranes. One type of organelle was similar to the thylakoid photosynthesizing organelles of blue-green algae (cyanobacteria). These findings indicate that the bodies may be a species of blue-green algae.
Serovars of Mycobacterium avium complex isolated from patients in Sweden
Authors: Hoffner S E , Källenius G , Petrini B , Brennan P J , Tsang A Y
Published: January 2020

The serovars of clinical isolates of Mycobacterium avium complex from 24 acquired immunodeficiency syndrome (AIDS) and 140 non-AIDS patients in Sweden were studied by using type-specific antisera. A wide distribution of serovars was seen. Serovar 6 was predominant in both groups of patients, isolated from 33 and 16% of the AIDS and non-AIDS patients, respectively. The results indicate geographical as well as disease-related differences in the distribution of M. avium complex serovars of clinical importance.
Diversity of rotavirus serotypes in Mexican infants with gastroenteritis
Authors: Padilla-Noriega L , Arias C F , López S , Puerto F , Snodgrass D R , Taniguchi K , Greenberg H B
Published: January 2020

One hundred thirty-two stool specimens from infants with rotavirus gastroenteritis hospitalized in two Mexican cities (Mexico City and Mérida) were examined by serotype- and subgroup-specific enzyme immunoassays. Among them, 38 (29%) were serotype 1, 15 (11%) were serotype 2, 13 (10%) were serotype 3, 22 (17%) were serotype 4, none was serotype 5 or 6, and 44 (33%) could not be serotyped. By subgrouping, 121 specimens were characterized as follows: 24 (18%) were subgroup 1, 97 (74%) were subgroup 2, and none had both subgroup specificities. While serotype 1 rotavirus predominated in the Mexico City area for 4 consecutive years (1984 to 1987), serotype 4 predominated in Mérida during the single epidemic season studied (1985). These data demonstrate that all four primary human rotavirus serotypes circulated in Mexico, with serotype 1 being the most prevalent. The seroneutralization responses of 14 of the 22 patients infected with serotype 4 strains had been previously studied. Of these 14 infants, 11 appeared to have primary infections, as indicated by absence of neutralizing antibodies in the acute-phase sera and their young age (8 months on average) at the time of illness. Seven patients seroresponded to serotypes 1 and 4; two seroresponded to serotypes 1, 3, and 4; three seroresponded to serotype 1; and two had low-level seroresponses to serotype 3 or 4. These data indicate that heterotypic neutralizing antibody responses occur frequently following infection with serotype 4 rotaviruses.
Pneumonia caused by a newly recognized pseudomonad in a child with chronic granulomatous disease
Authors: Trotter J A , Kuhls T L , Pickett D A , Reyes de la Rocha S , Welch D F
Published: January 2020

A pseudomonad was isolated from the pleural fluid and pulmonary decortication tissue of a 5-year-old child with chronic granulomatous disease. Although the isolate was phenotypically similar to Pseudomonas cepacia, its biochemical profile was more similar to that of Pseudomonas pickettii biovar 2. Its slow growth rate, ability to hydrolyze urea rapidly, and lateral and polar flagellar pattern were suggestive of Oligella ureolytica (formerly CDC group IVe). The cellular fatty acid composition was similar to that of P. cepacia and Pseudomonas gladioli, except for the presence of dodecanoic acid. Numerical analysis of the fatty acid data supported the interrelatedness of the isolate with other species of the pseudomallei group (rRNA homology group II) of Pseudomonas. The organism described in this report is an addition to the growing list of catalase-positive organisms which can potentially cause severe morbidity in patients with chronic granulomatous disease.
Occurrence of nonspecific reactions among stool specimens tested by the Abbott TestPack rotavirus enzyme immunoassay
Authors: Lipson S M , Leonardi G P , Salo R J , Schutzbank T E , Kaplan M H
Published: January 2020

Sixty-five stool specimens obtained from children suffering from gastroenteritis were tested for the presence of antigen to rotavirus by the Abbott TestPack Rotavirus (TestPack) enzyme immunoassay kit. The Kallestad Pathfinder enzyme immunoassay, polyacrylamide gel electrophoresis, immune electron microscopy, and virus isolation were utilized as reference assays. Fifty-four specimens were in accord by TestPack and Kallestad Pathfinder. Among 11 discordant specimens positive with TestPack but negative by Kallestad Pathfinder, rotavirus was not identified by polyacrylamide gel electrophoresis, immune electron microscopy, or isolation in primary African green monkey kidney cell cultures. TestPack displayed a performance specificity of 83%. The inordinately high number of stool specimens reported as false-positive by TestPack precludes the incorporation of this antigen detection kit into our routine regimen of diagnostic virologic testing.
Accuracy and reproducibility of the MicroScan rapid anaerobe identification system with an automated reader
Authors: Stoakes L , Kelly T , Manarin K , Schieven B , Lannigan R , Groves D , Hussain Z
Published: January 2020

Rapid anaerobe identification (MicroScan) panels (4 h) were evaluated both visually and by the AutoScan-4, a computer-controlled microplate reader. The results of both reading methods were compared with identifications obtained by the conventional (Virginia Polytechnic Institute) method. In total, 237 anaerobes were tested. Correct identifications were obtained for 166 strains (70%) by visual reading and 157 strains (66.2%) by the AutoScan-4. Supplementary tests resulted in 80.1 and 76.7% total correct identifications, respectively. Comparison of the two reading methods revealed complete agreement for 169 strains. Differences between the two reading methods were due to difficulties in reading specific reactions. This was especially true with the clostridial species. The performance of the MicroScan system in the identification of anaerobic bacteria appears comparable to that of other 4-h identification systems for anaerobes, but this system shows significant variance from the conventional system. Improvements in the trays and data base are required before the system can be recommended for routine use.
Phenotypic conversion of Pseudomonas aeruginosa in cystic fibrosis
Authors: Fegan M , Francis P , Hayward A C , Davis G H , Fuerst J A
Published: January 2020

Pseudomonas aeruginosa strains isolated from cystic fibrosis patients were tested for production of exoenzymes, sensitivity to pooled normal human serum, and colony morphology. Strains isolated from patients exhibiting a severe form of the disease were seen to produce a decreased range of exoenzymes, to show an increase in their serum sensitivity, and to be predominantly mucoid in colonial character compared with strains isolated from patients with a milder form of the disease. These results suggest that P. aeruginosa undergoes phenotypic changes with respect to exoenzyme secretion, serum sensitivity, and colony form as the clinical condition of the cystic fibrosis patient changes.
Use of two-dimensional gas chromatography with electron-capture detection for the measurement of lipopolysaccharides in peritoneal fluid and plasma from rats with induced peritonitis
Authors: Sonesson A , Larsson L , Andersson R , Adner N , Tranberg K G
Published: January 2020

The content of 3-hydroxymyristic acid from Escherichia coli lipopolysaccharide in peritoneal fluid and plasma from rats was determined by two-dimensional gas chromatography with electron-capture detection of the 3-O-pentafluorobenzoyl methyl ester derivative. The detection limit of lipopolysaccharide in peritoneal fluid was 3 ng/ml. An experimental model of E. coli peritonitis in the rat was used, with and without coinjection of bile. The concentrations of lipopolysaccharide were highest in both peritoneal fluid and plasma samples from rats injected with E. coli and bile, reaching a maximum 1 h after injection by the gas chromatographic method. Corresponding Limulus assay results for peritoneal samples showed a small increase of lipopolysaccharide concentrations during the first 4 h after injection, followed by a substantial increase. The results indicate that bile salts cause an increased release of lipopolysaccharide from gram-negative bacterial cells in vivo and that this may be responsible for the high mortality caused by peritonitis. In contrast to the Limulus assay, gas chromatography enables the total amount of lipopolysaccharide in a clinical sample to be determined.
Immunodominant epitopes of the adhesin of Mycoplasma pneumoniae
Authors: Jacobs E , Pilatschek A , Gerstenecker B , Oberle K , Bredt W
Published: January 2020

Overlapping octapeptides from the amino acid sequence of the adherence protein of Mycoplasma pneumoniae were synthesized and used as the antigen in an enzyme-linked immunosorbent assay with serum samples from M. pneumoniae-infected patients. Of a sequence of 338 amino acids positioned between leucine 801 and leucine 1139, only two regions with immunodominant continuous epitopes were detected. The immunoglobulin G and immunoglobulin M antibodies of child and adult patients reacted especially with the NH2-(810)-W-I-G-N-G-Y-R-Y peptide but also reacted with the NH2-(1124)-F-T-D-F-V-K-P-R peptide.
Molecular identification by RNA-RNA hybridization of a human rotavirus that is closely related to rotaviruses of feline and canine origin
Authors: Nakagomi O , Ohshima A , Aboudy Y , Shif I , Mochizuki M , Nakagomi T , Gotlieb-Stematsky T
Published: January 2020

With a few exceptions subgroup I group A human rotavirus strains have short RNA patterns, whereas most animal rotavirus strains belong to subgroup I and have long RNA patterns. Thus, new isolates of subgroup I human rotaviruses with long RNA patterns are considered to have a high likelihood of being animal rotaviruses. A group of human rotaviruses represented by the AU-1 strain has recently been shown to be genetically related to a feline rotavirus (FRV-1) isolated in Japan. A human rotavirus, strain Ro1845, which is similar to the AU-1 strain in its subgroup (I), serotype (3), and electropherotype (long), was compared with various human and animal strains by RNA-RNA hybridization to determine its genogroup, a term proposed to classify rotaviruses based on their gene homology. The Ro1845 strain did not show a significant level of homology with AU-1, FRV-1, or other human strains, indicating that the Ro1845 strain is different in its genogroup not only from the AU-1 strain but also from other human strains. However, the Ro1845 strain showed a high degree of homology with another feline rotavirus (Cat97) isolated previously in Australia, suggesting that the Ro1845 strain might originate from a feline rotavirus that is genetically distinct from the Japanese FRV-1 strain. Furthermore, the Ro1845 strain as well as the Cat97 strain were related genetically to the canine rotavirus RS15 strain. Taken together, these results indicate that at least two genogroups are present in feline rotaviruses, one resembling the AU-1 strain and the other resembling the Ro1845 strain as well as canine rotaviruses.
Use of a novel phosphatase test for simplified identification of species of the tribe Proteeae
Authors: Pompei R , Cornaglia G , Ingianni A , Satta G
Published: January 2020

Phosphatase activity of 334 isolates of the tribe Proteeae carefully identified to species level has been evaluated by using both a number of conventional tests and three versions of the novel methyl green-phenolphthalein (MGP) method (G. Satta. R. Pompei, G. Grazi, and G. Cornaglia, J. Clin. Microbiol. 26:2637-2641, 1988). We found that the different species of Proteeae show different and easily distinguishable. behaviors by the MGP method, while all of them behave in a uniform way in the conventional tests. On studying the mechanism underlying these different behaviors, we found that the phosphatase activity of Morganella morganii and Providencia stuartii is higher than that of all other members of the family Enterobacteriaceae. Furthermore, we found that all P. stuartii strains display the property (shared by virtually none of the other species of Enterobacteriaceae) of excreting significant amounts of phosphatase. This extracellular activity is reliably detected by the MGP method but not by conventional tests. Finally, by exploiting the peculiar phosphatase activity of this tribe, as revealed by the MGP method, we devised a simple scheme for routine identification of the species of Proteeae and evaluated its reliability by comparing it with three commonly used commercial kits. The new scheme proved much simpler, but also more reliable, since identifications obtained by this method were in almost complete accord (99%) with those of the reference identification schemes, while in the commercial systems examined the percentage of errors ranged from 13 to 14 in the identification of Providencia species.
Plasmid profiling of members of the family Enterobacteriaceae, lactobacilli, and bifidobacteria to study the transmission of bacteria from mother to infant
Authors: Tannock G W , Fuller R , Smith S L , Hall M A
Published: January 2020

Plasmid profiles of isolates of the family Enterobacteriaceae, lactobacilli, and bifidobacteria cultured from vaginal, oral, and rectal swabs collected from women soon after admission to a maternity hospital were compared with those of strains detected in the feces of their infants. Lactobacilli inhabiting the vaginas of the mothers did not appear to colonize the infant digestive tract, but evidence for the transmission of fecal isolates of the Enterobacteriaceae and bifidobacteria from mother to infant was obtained in four out of five cases. Many of the bifidobacteria isolated were plasmid-free but could be distinguished with biochemical profiles.
Computer-assisted methods for assessing strain relatedness in Candida albicans by fingerprinting with the moderately repetitive sequence Ca3
Authors: Schmid J , Voss E , Soll D R
Published: January 2020

When used to probe EcoRI-digested Candida albicans DNA, the moderately repetitive sequence Ca3 generated a Southern blot hybridization pattern which included 15 to 25 bands, depending upon the strain. The pattern was stable through 400 generations in each of three independent strains but variable between most of 46 unrelated tester strains, making it a very effective probe for discrimination between strains. Computer-assisted methods (Dendron) were developed for storage of Ca3 patterns in data files, calculation of similarity (SAB) values between strains based upon band positions and intensities, and generation of histograms and dendrograms based on SAB values for all strains or any subset of strains in large epidemiological studies. In testing the effectiveness of the system, it was found that (i) multiple isolates from different body locations of the same healthy individual could represent either the same strain or different strains, (ii) isolates from oral lesions of a husband and wife represented the same strain, (iii) strains isolated from the mouths of 10 healthy individuals on the same day and in the same geographical location were as dissimilar on average as the 46 unrelated tester strains, and (iv) strains isolated from seven immunocompromised patients hospitalized over a 2.5-month period in the same hospital were highly similar, indicating nosocomial origin. The apparent effectiveness of these fingerprinting methods and the Dendron program suggests that interlaboratory procedures for fingerprinting should be standardized and all patterns should be analyzed and stored in a common and accessible data base for broad epidemiological analysis.
Antigenic characterization of small, round-structured viruses by immune electron microscopy
Authors: Okada S , Sekine S , Ando T , Hayashi Y , Murao M , Yabuuchi K , Miki T , Ohashi M
Published: January 2020

Small, round-structured viruses (SRSVs) detected from nonbacterial gastroenteritis outbreaks in Tokyo and Saitama Prefecture, Japan, during the period from 1977 to 1988 were tentatively classified into nine antigenic patterns from SRSV-1 (S-1) to SRSV-9 (S-9) by cross-immune electron microscopy (IEM). S-1 and S-2 appeared pattern specific, while S-3 to S-9, distinguishable from each other in their reactivity, appeared somewhat antigenically related. Their antigenic relatedness to the Norwal, Hawaii, and Otofuke agents was also examined by IEM by using antisera to these agents. S-3 appeared most closely related to the Norwalk agent. S-4 and S-5 were related to the Norwalk agent and, presumably, were distantly related to the Hawaii and Otofuke agents. S-6 and S-7 were related to the Hawaii and Otofuke agents. S-8 and S-9 were related to the Otofuke agent and, presumably, were distantly related to the Hawaii agent. The prevalence of each antigenic pattern in 38 outbreaks was examined: S-8 was implicated in 24% of the outbreaks S-5 in 16%, S-4 in 13%, S-9 in 13%, S-6 in 11%, and others in 5%.
Restriction endonuclease analysis of Eikenella corrodens
Authors: Chen C K , Sunday G J , Zambon J J , Wilson M E
Published: January 2020

Eikenella corrodens is a gram-negative facultative bacillus commonly found in the oral cavity. Although the role of E. corrodens in periodonititis is not clear, its isolation from extraoral infections attests to its pathogenic potential. Previous studies suggested that this species is phenotypically diverse. In the present study, we used restriction endonuclease analysis (REA) to assess the genetic diversity of this species and to explore the applicability of REA in studying the transmission of E. corrodens. Two groups of E. corrodens isolates were used in this study. Group 1 included 47 epidemiologically independent isolates recovered from dental plaques in periodontally healthy subjects and periodontitis patients and from extraoral infections in different geographic areas. Group 2 E. corrodens included 40 isolates recovered from two periodontitis patients and two periodontally healthy subjects. The results indicated that E. corrodens is genetically heterogeneous, as determined by REA. The majority of the group 1 E. corrodens isolates exhibited strain-specific restriction patterns. Forty restriction patterns were distinguishable among the 47 isolates. Analyses of group 2 isolates revealed that three of four subjects harbored more than one clonal type of E. corrodens. In one instance, a periodontitis patient was found to be colonized by six different clones. Furthermore, two different clonal types of E. corrodens were recovered from a single periodontal pocket in this patient. The results indicated that REA may be a useful tool in the epidemiologic investigation of E. corrodens infections.
DNA probe culture confirmation assay for identification of thermophilic Campylobacter species
Authors: Tenover F C , Carlson L , Barbagallo S , Nachamkin I
Published: January 2020

We studied the ability of a new DNA probe-based assay system to correctly identify isolates of the thermophilic campylobacters Campylobacter jejuni, C. coli, and C. laridis grown in vitro. We examined 424 organisms, including 214 Campylobacter isolates and 210 other aerobic and anaerobic isolates. The probe assay, which uses a new homogeneous system in which all reactions take place within a single tube, demonstrated 100% accuracy, producing neither false-positive nor false-negative results. The assay does not, however, distinguish among C. jejuni, C. coli, and C. laridis.
Isolation, characterization, and host-cell-binding properties of a cytotoxin from Campylobacter jejuni
Authors: Mahajan S , Rodgers F G
Published: January 2020

A 68,000-molecular-weight protein was isolated by polyacrylamide gel electrophoresis from the organism-free filtrate of a fully virulent clinical strain of Campylobacter jejuni. The eluted protein was heat labile, was inactivated at either pH 3.0 or 9.0, was sensitive to trypsin, and was lethal for fertile chicken eggs. It also had toxic effects on chicken embryo fibroblast, Chinese hamster ovary (CHO), and intestinal 407 (Int407) cells. A monoclonal antibody (CETPMAb4) raised to this eluted toxic protein (ETP) from C. jejuni abolished these toxic activities. Homology between C. jejuni ETP and Vibrio cholerae toxin was not observed in that specific antisera to each did not block their respective toxic activities. In enzyme-linked immunosorbent assays, ETP, unlike chlorea enterotoxin, did not bind to GM1 ganglioside. Furthermore, the C. jejuni toxin had cytotoxinlike properties and induced rounding of CHO cells. Binding of ETP to Int407 and primary chicken embryo fibroblast cells was maximal after 2 h as assessed by enzyme-linked immunosorbent assay, and this toxin adherence to host cell membranes was significantly reduced by prior treatment of the cells with proteolytic enzymes, neuraminidase, or glutaraldehyde but not by treatment with beta-galactosidase, lipase, Nonidet P-40, or sodium metaperiodate. In competitive binding assays, sugars, lectins, or GM1 ganglioside did not adversely influence uptake of ETP by these cells. These results suggest that the ETP produced by C. jejuni is a cytotoxin which binds to Int407 cells via a protein- or glycoproteinlike receptor on cell membranes and possesses properties dissimilar to those of V. cholerae toxin.
Reactivity of human Lyme borreliosis sera with a 39-kilodalton antigen specific to Borrelia burgdorferi
Authors: Simpson W J , Schrumpf M E , Schwan T G
Published: January 2020

Borrelia burgdorferi is the causative agent of Lyme borreliosis, a spirochetal illness with a variety of acute clinical manifestations that may lead to debilitating neurological and arthritic complications. Diagnosis is difficult because symptoms mimic a variety of unrelated clinical conditions, spirochetes cannot always be isolated from infected patients, and current serological tests are frequently inconclusive because of the presence of cross-reacting non-B. burgdorferi antibodies. To identify antigens specific to B. burgdorferi that could be used in the serodiagnosis of Lyme borreliosis, we screened a Borrelia DNA expression library in Escherichia coli for antigens reactive with human Lyme borreliosis sera. One clone carried a 6.3-kilobase EcoRI chromosomal fragment (pSPR33), which encoded two species-specific antigens with molecular masses of 28 (P28) and 39 (P39) kilodaltons (kDa). These two antigens were immunologically distinct from OspA, OspB, and the 41-kDa flagellin. Ninety-four serum specimens from patients having Lyme borreliosis were tested for reactivity with P39. All of 33 the serum specimens with immunofluorescence assay titers of greater than or equal to 1:256, 13 of 17 serum specimens with titers of 1:128, and 14 of 44 serum specimens with titers of less than or equal to 1:64 reacted with P39. Notably, many sera reactive to P39 did not appear to react with the 41-kDa flagellin. Therefore, antibody to P39 could be mistaken for antibody to the 41-kDa flagellin in tests of human sera by Western blot (immunoblot). Twenty-five control serum specimens, which included sera from syphilitic, relapsing fever, and amyotrophic lateral sclerosis patients as well as from 10 normal individuals, did not react to P39. Our data suggest that P39 may be a useful antigen for the serological confirmation of Lyme borreliosis.
Epidemiological markers for epidemic strain and carrier isolates in an outbreak of nosocomial oxacillin-resistant Staphylococcus aureus
Authors: Bouvet A , Fournier J M , Audurier A , Branger C , Orsoni A , Girard C
Published: January 2020

An outbreak of nosocomial infections occurring in a postoperative intensive care unit was caused by a single strain of oxacillin-resistant Staphylococcus aureus. Six patients were infected, or colonized, by this strain, which was traced by using the following four epidemiological markers: antibiogram, bacteriophage type, capsular polysaccharide type, and esterase electrophoretic type. This strain was compared with S. aureus isolates obtained from the noses of 13 carriers from a group of 42 staff members. A good correlation in terms of phenotypic markers was found between the epidemic strain and a strain isolated from one carrier. Both exhibited the same pattern of multiple resistance as well as the same phage type, 77, capsular polysaccharide type, 5, and esterase electrophoretic type, 6. In contrast, an oxacillin-resistant strain, isolated from another carrier, differed from the epidemic strain by susceptibility to rifampin and by susceptibility to four additional bacteriophages. The other 11 strains isolated from carriers were susceptible to oxacillin and exhibited widely different phenotypes. These results confirm the interest of using several epidemiological markers to trace the spread of epidemic S. aureus strains and to delineate the carrier strains.
Comparison of rotavirus immunoglobulin A coproconversion with other indices of rotavirus infection in a longitudinal study in childhood
Authors: Coulson B S , Grimwood K , Masendycz P J , Lund J S , Mermelstein N , Bishop R F , Barnes G L
Published: January 2020

In order to determine the sensitivity and reliability of antirotaviral fecal immunoglobulin A (IgA) as an indicator of rotavirus reinfection, the antibody responses to rotavirus of 44 infants with severe rotavirus gastroenteritis recruited on admission to a hospital were studied. Feces were collected daily during hospitalization and weekly thereafter, and sera were obtained every 4 months, for 6 to 32 months (median, 17 months). Antirotaviral IgG, IgA, and IgM were measured by enzyme immunoassay in all samples. Rotavirus antigen, rotavirus-neutralizing antibody, and total IgA were measured in feces. The results showed that use of an IgA index (ratio of specific IgA to total IgA) was unnecessary to identify copro-IgA conversion to rotavirus. The other markers of rotavirus infection tested showed a high level of predictive accuracy of coproconversion in rotavirus-neutralizing antibody. Copro-IgM, serum IgM, and virus in feces were insensitive measures of neutralizing antibody coproconversion. Seroconversion in IgG or IgA was detected in 46% of neutralizing coproconversions. The most sensitive marker, present in 92% of neutralizing coproconversions, was antirotaviral fecal IgA conversion. This correlation of fecal IgA with fecal neutralizing antibody suggests that coproconversions in IgA represent true elevations in antirotaviral IgA with neutralizing capacity. A coproconversion in IgA appears to indicate genuine rotavirus infection. Copro-IgA conversions in feces collected weekly are likely to be more sensitive markers of rotavirus reinfection than are seroconversion and virus detection combined in epidemiological studies of acute diarrhea in children and in rotavirus vaccine trials.
Immunologic abnormalities in chronic fatigue syndrome
Authors: Klimas N G , Salvato F R , Morgan R , Fletcher M A
Published: January 2020

The chronic fatigue syndrome (CFS), formerly known as chronic Epstein-Barr virus syndrome, is a clinical state of some complexity and uncertain etiology. In order to characterize in a comprehensive manner the status of laboratory markers associated with cellular immune function in patients with this syndrome, 30 patients with clinically defined CFS were studied. All of the subjects were found to have multiple abnormalities in these markers. The most consistent immunological abnormality detected among these patients, when compared with normal controls, was low natural killer (NK) cell cytotoxicity. The number of NK cells, as defined by reactivity with monoclonal antibody NKH.1 (CD56), was elevated, but the killing of K562 tumor cells per CD56 cell was significantly diminished. Lymphoproliferative responses after stimulation with phytohemagglutinin and pokeweed mitogen were decreased in most patients when compared with those in normal controls, as was the production of gamma interferon following mitogen stimulation. Lymphocyte phenotypic marker analysis of peripheral blood lymphocytes showed that there were significant differences between patients with CFS and controls. There was an increase in the percentage of suppressor-cytotoxic T lymphocytes, CD8, and a proportionally larger increase in the number of CD8 cells expressing the class II activation marker. Most patients had an elevated number of CD2 cells which expressed the activation marker CDw26. The numbers of CD4 cells and the helper subset of CD4+CD29+ cells in patients with CFS were not different from those in controls. There was, however, a significant decrease in the suppressor inducer subset of CD4+ CD45RA+ cells. The number of B cells, CD20 and CD21, were elevated, as were the numbers of a subset of B cells which coexpressed CD20 and CD5. The patterns of immune marker abnormalities observed was compatible with a chronic viral reactivation syndrome.
Evaluation of a new bicolored latex agglutination test for immunological diagnosis of hepatic amoebiasis
Authors: Robert R , Mahaza C , Bernard C , Buffard C , Senet J M
Published: January 2020

A new bicolored latex agglutination amoeba test (BLA) for detection of antibodies against Entamoeba histolytica was evaluated for its practicability and diagnostic sensitivity and specificity. BLA is rapid (5 min) and simple to perform. It requires only 20 microliters of a 1/3-diluted serum, 17 microliters of reagent, and a glass slide. Reading of the test is easy because a positive result shows a green spot with a red surrounding edge. This bicolored pattern is easily distinguishable from the negative test result, which shows a homogeneous dark-brown spot. By using serum samples from 348 individuals, BLA was compared with immunofluorescence assay, indirect hemagglutination, and counterimmunoelectrophoresis. Sensitivity, specificity, efficiency, and positive and negative predictive values of the four methods were almost identical. The results of this study indicate that BLA could be very useful both as a screening method for the diagnosis of invasive amoebiasis and for epidemiological purposes.
Comparison of three enzyme immunoassays to tissue culture for the diagnosis of rotavirus gastroenteritis in infants and young children
Authors: Christy C , Vosefski D , Madore H P
Published: January 2020

Three enzyme immunoassays (EIAs), Rotazyme II, IDL, and Pathfinder, were evaluated for rotavirus detection in stool and rectal swab specimens from children with symptomatic gastroenteritis and compared with virus isolation in primary African green monkey kidney cells. Of 125 specimens tested, 49 were rotavirus positive by tissue culture isolation; of these 49, 40 were positive by Rotazyme II, 43 were positive by IDL, and 46 were positive by Pathfinder EIAs. As compared with tissue culture isolation, the Rotazyme II, IDL, and Pathfinder EIAs had sensitivities of 82, 88, and 94%, specificities of 90, 99, and 95%, and overall agreements of 86, 94, and 94%, respectively.
Sialidase activity of the "Streptococcus milleri group" and other viridans group streptococci
Authors: Beighton D , Whiley R A
Published: January 2020

Viridans group streptococci were examined for the production of sialidase (neuraminidase) activity, using the fluorescent substrate 4-methylumbelliferyl-alpha-D-N-acetylneuraminic acid in a simple and rapid (15-min) assay. Sialidase was produced by all strains of Streptococcus oralis and S. intermedius and by a majority of S. mitis strains. S. mutans, S. sobrinus, S. gordonii, S. sanguis, S. vestibularis, S. salivarius, S. anginosus, S. constellatus, "S. parasanguis," and the "tufted fibril group" were uniformly negative. Sialidase production may be a useful characteristic to assist in the identification of viridans group streptococci.
Presence of gamma interferon in human acute and congenital toxoplasmosis
Authors: Raymond J , Poissonnier M H , Thulliez P H , Forestier F , Daffos F , Lebon P
Published: January 2020

The production of gamma interferon in acute acquired and congenital toxoplasmosis was studied. Gamma interferon was produced at significant titers (P less than 0.001) in the course of both congenital toxoplasmosis and acquired toxoplasmosis at an early stage of infection, when Toxoplasma gondii was multiplying. Its presence in fetal blood was correlated with the positive inoculation of fetal blood or amniotic fluid into mice (95%). The data suggest that the fetus is able to synthesize gamma interferon as early as week 21 of pregnancy. This test, easily and rapidly performed, could be included among those useful for diagnosing fetal toxoplasmic disease.
Selective medium for Pasteurella multocida and its use to detect oropharyngeal carriage in pig breeders
Authors: Avril J L , Donnio P Y , Pouedras P
Published: January 2020

A selective culture medium for isolation of Pasteurella multocida was prepared by incorporating 2 mg of amikacin per liter, 4 mg of vancomycin per liter, and 4 mg of amphotericin B per liter into Mueller-Hinton blood agar. Use of this medium revealed the presence of P. multocida in the oropharynges of 19 of the 49 pig breeders who were examined.
Comparison of IDEIA III and cell culture for the detection of Chlamydia trachomatis in endocervical specimens
Authors: Lebar W D , Schubiner H , Jemal C , Herschman B R
Published: January 2020

A total of 803 endocervical samples were obtained from females with clinical or epidemiological histories suggesting chlamydia infection. These specimens were tested by IDEIA III and cell culture for the presence of Chlamydia trachomatis. After resolution of discrepant results by direct fluorescent-antibody staining of pelleted cell culture transport materials, IDEIA III demonstrated sensitivity, specificity, and positive and negative predictive values of 93.8, 99, 92.9, and 99.1%, respectively.
Cord formation in BACTEC 7H12 medium for rapid, presumptive identification of Mycobacterium tuberculosis complex
Authors: Yagupsky P V , Kaminski D A , Palmer K M , Nolte F S
Published: January 2020

We evaluated cord formation in BACTEC 7H12 medium as a criterion for rapid identification of Mycobacterium tuberculosis complex. Kinyoun-stained smears, prepared from 270 radiometrically positive BACTEC 7H12 bottles, were examined independently by three observers. Smears from 93.2, 88.6, and 83.0% of the M. tuberculosis complex cultures were read as cord positive, and smears from 97.3, 97.8, and 99.5% of the mycobacteria other than M. tuberculosis cultures were read as cord negative by the three observers, respectively. There was 93.3% agreement between the observers. The presence of cords in BACTEC 7H12 medium can be a reliable criterion for rapid, presumptive identification of M. tuberculosis complex.
Cell culture propagation of a coronavirus isolated from cows with winter dysentery
Authors: Benfield D A , Saif L J
Published: January 2020

Fecal filtrates from cows with winter dysentery were inoculated into gnotobiotic and conventional calves, and a coronavirus was isolated from calf feces. Cytopathic effects were observed on human rectal tumor cells but not bovine cell cultures. The winter dysentery isolates morphologically and antigenically resembled the Mebus strain of bovine coronavirus.
Evaluation of the Becton Dickinson Directigen test for respiratory syncytial virus in nasopharyngeal aspirates
Authors: Kok T , Barancek K , Burrell C J
Published: January 2020

A premarket trial of the Becton Dickinson Directigen respiratory syncytial virus membrane-based enzyme immunoassay compared the test with virus isolation for the detection of respiratory syncytial virus in 583 nasopharyngeal aspirates. After modification, the Directigen test showed a sensitivity of 83% and a specificity of 90%. It offers the potential for an efficient bedside test--without the need for any equipment--for the diagnosis of respiratory syncytial virus infection and requires only a 0.25-ml sample volume. However, for optimum reliability, freezing-thawing of samples and access to a confirmatory test were shown to be necessary.
Microplate hybridization of amplified viral DNA segment
Authors: Inouye S , Hondo R
Published: January 2020

We have developed a simple hybridization method for a DNA segment which is amplified by the polymerase chain reaction: after heat denaturation, the amplified DNA segment with a length of more than 300 bases is adsorbed to microplate wells in the presence of 1.5 M NaCl or 0.5 M ammonium sulfate; the immobilized DNA is hybridized with a biotin-labeled DNA probe; then, the hybridization signal is detected by streptavidin-conjugated beta-galactosidase or peroxidase. This method has several advantages over the conventional dot blot hybridization method: (i) radioisotopes are not used, (ii) synthetic oligonucleotide for the probe is not needed, (iii) the time required for washing of the solid phase is greatly reduced, and (iv) the baking and prehybridization procedures are eliminated. By this method, we were able to detect viral genomes in vesicle specimens from patients infected with varicella-zoster virus.