bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of the enhanced rapid identification method for Gardnerella vaginalis
Authors: Lien E A , Hillier S L
Published: January 2020

The enhanced rapid identification method (RIM; Austin Biological Laboratories), a micromethod for the identification of Gardnerella vaginalis, is based on starch and raffinose fermentation and hippurate hydrolysis. We tested 105 clinical isolates of G. vaginalis with both the RIM and standard biochemical tests. The RIM agreed with the standard biochemical methods for 96 (91.4%) of the strains; nine isolates which were hippurate hydrolysis positive by standard biochemical tests were hippurate hydrolysis negative in the RIM. RIM may serve as a useful adjunct to Gram stain and colony morphology for the identification of G. vaginalis.
Catheter-related bacteremia caused by Oerskovia turbata
Authors: LeProwse C R , McNeil M M , McCarty J M
Published: January 2020

A case of Broviac catheter-related bacteremia in a 3-year-old boy infected with Oerskovia turbata is discussed. The blood cultures of the patient remained positive despite antibiotic therapy, and only with catheter removal was his blood sterilized. The microbiology and antimicrobial susceptibilities of this organism are described.
Equine zygomycosis caused by Conidiobolus lamprauges
Authors: Humber R A , Brown C C , Kornegay R W
Published: January 2020

A 15-year-old Arabian mare from southern Louisiana with a 2-month history of periodic epistaxis and severe weight loss had a large, fibrosing, granulomatous mass containing numerous nodules ("kunkers") projecting dorsally into the nasopharynx, and was euthanized at the owner's request. In addition to these kunkers, the mass contained a single trematode tentatively identified as Fasciola hepatica. Several kunkers were removed, washed thoroughly in sterile water and embedded in nutrient agars; the fungus that grew out of them was identified as Conidiobolus lamprauges Drechsler (Entomophthorales: Ancylistaceae). This is the first report of C. lamprauges from any vertebrate mycosis, and only the third Conidiobolus species reported from vertebrates. Unlike many other entomophthoraleans, the fungus isolated from this mycosis grew well at 37 degrees C. The possible means by which Conidiobolus species may infect vertebrates is discussed. These mycoses probably result most often from chronic exposure during sleep to conidia discharged from fungal growth on decaying plant material in the bedding.
Comparison of bacteria with and without plasmid-encoded proteins as antigens for measurement of immunoglobulin M, G, and A antibodies to Yersinia enterocolitica by enzyme-linked immunosorbent assay
Authors: Granfors K , Lahesmaa-Rantala R , Ståhlberg T H , Toivanen A
Published: January 2020

Yersinia enterocolitica serovar O:3 bacteria with and without plasmid-encoded proteins were compared as antigens in an enzyme-linked immunosorbent assay. Good correlations between the two antigen preparations were obtained for immunoglobulin M (IgM), IgG, and IgA antibodies of patients with yersiniosis. For routine diagnostic purposes, these antigens are considered equal.
Absence of monocyte procoagulant activity during the immune response to influenza virus
Authors: Mathies M , Hogg N
Published: January 2020

The ability of mononuclear phagocytes (MPh) to manifest procoagulant activity (PCA) resulting in the formation of fibrin is thought to be a key MPh effector function in tissue repair. The present study addresses the question of whether monocyte PCA is confined to tissue hypersensitivity reactions or is a general correlate of all immune responses. We show here that PCA is not the obligate outcome when the immune system is stimulated. In particular, under in vitro conditions in which a mitogen (phytohemagglutinin) or an antigen (purified protein derivative of tuberculin) elicits good PCA responses, incubation with influenza virus does not result in the generation of PCA, although other parameters of response to the virus appear to be intact. Moreover, influenza virus can cause suppression of PCA when cultures are stimulated with either phytohemagglutinin, purified protein derivative of tuberculin, or endotoxin, conditions which would otherwise result in good PCA responses. This lack of PCA persists throughout the culture period and is not caused by an effect of influenza virus on the viability of either MPh or leukocytes in general.
Clonal groups of Salmonella typhimurium in New York State
Authors: McDonough P L , Timoney J F , Jacobson R H , Khakhria R
Published: January 2020

The epidemiology of 278 strains of Salmonella typhimurium isolated from 1973 to 1981 from animals in New York State was studied by using four "fingerprinting" techniques, bacteriophage type (B.R. Callow, J. Hyg. 57:346-359, 1959), biotype (J. P. Duguid, E. S. Anderson, G. A. Alfredsson, R. Barker, and D. C. Old, J. Med. Microbiol. 8:149-166, 1975), plasmid profile, and antibiogram. Phage type with biotype was the most useful marker for distinguishing clonal groups of S. typhimurium. Four clones of S. typhimurium predominated, i.e., phage type/biotypes U275/26, 49/26, 10/3, and 2/3. U275/26 and 49/26 were commonly found until 1976, but clones 10/3 and 2/3 were predominant after 1976. Comparison of results with data from Canada suggested a dissemination of strains of S. typhimurium between Canada and New York. Cattle were a common source of phage type 49, as has been observed in other countries.
DNA hybridization probe for clinical diagnosis of Entamoeba histolytica
Authors: Samuelson J , Acuna-Soto R , Reed S , Biagi F , Wirth D
Published: January 2020

As an alternative to microscopic identification of Entamoeba histolytica parasites isolated from stool, a sensitive and species-specific DNA hybridization probe was made for rapid diagnosis of E. histolytica parasites in clinical samples directly applied to nylon membranes. The DNA hybridization probe was made by screening a genomic library of a virulent HM-1:IMSS strain of E. histolytica to detect recombinant plasmids containing highly repeated parasite DNA sequences. Four plasmid clones that reacted across Entamoeba species coded for highly repeated rRNA genes of E. histolytica. Four other plasmid clones were E. histolytica specific in that they bound to four axenized and nine xenic strains of E. histolytica but did not recognize closely related E. histolytica-like Laredo, Entamoeba moshkovskii, or Entamoeba invadens parasites. The diagnostic clones detected as few as eight cultured amoebae and did not distinguish between pathogenic and nonpathogenic zymodemes of E. histolytica. The diagnostic clones were sequenced and contained 145-base-pair sequences which appear to be tandemly repeated in the genome. No stable transcript which is homologous to the diagnostic DNA was detected. In a study of stool samples from Mexico City shown by microscopy to contain E. histolytica, Entamoeba coli, Giardia lamblia, Endolimax nana, Trichuris trichiuria, and Chilomastix mesnili parasites, the DNA hybridization probe demonstrated a sensitivity of 1.0 and a specificity of 0.93. We conclude that the DNA hybridization probe can be used for rapid and accurate diagnosis of E. histolytica parasites.
Production of bacteriocin-like antagonism by clinical isolates of Yersinia enterocolitica
Authors: Cafferkey M T , McClean K , Drumm M E
Published: January 2020

Fourteen clinical isolates of Yersinia enterocolitica serotype O:3 and four well-documented virulent strains of serotypes O:3, O:8, and O:9 were biotyped and examined for plasmid-associated autoagglutination and calcium dependency and for epithelial cell adherence. These strains were tested for the production of bacteriocin-like antagonism by using tryptone soya blood agar at room temperature and at 37 degrees C. By using the cross-streaking method, three clinical isolates produced inhibitory substances at room temperature. These substances were active against a variety of clinical isolates and their plasmid-cured derivatives at both room temperature and 37 degrees C. The inhibition was easier to read after incubation of the cross-streaked plate at 37 degrees C. The inhibition patterns indicate that two of the three producer strains appear to recognize potentially virulent O:3 strains, with or without the virulence plasmid.
Switching of Candida albicans during successive episodes of recurrent vaginitis
Authors: Soll D R , Galask R , Isley S , Rao T V , Stone D , Hicks J , Schmid J , Mac K , Hanna C
Published: January 2020

Strain relatedness and switching were monitored in Candida albicans strains isolated from different body locations through three episodes of recurrent vulvovaginal candidiasis separated by two treatment-latency periods in a single patient. Strain relatedness was assessed by comparing Southern blot hybridization patterns with the relatively immobile mid-repeat sequence Ca3. The following conclusions are demonstrated. (i) Three different strains of C. albicans colonized the mouth, the area under the breasts, and the vulvovaginal, anal, and rectal regions, respectively, at the time of the first infection. (ii) The same strain of C. albicans was responsible for the three vaginal infections. (iii) Switching of colony phenotype occurred with each new vaginal infection. (iv) Enrichment of drug-resistant switch phenotypes (assessed in vitro) was unlikely the basis for the changes in the switch phenotypes of the strain found in the vulvovaginal, anal, and rectal areas after treatment of the first infection with clotrimazole. (v) The same strain of C. albicans was responsible for the recurrent increases in mouth colonization and was distinct from the recurrent vaginal strain. The results of this case study demonstrate the need for further detailed analyses of full-body mycofloras, strain relatedness, switching repertoires, and changes in drug susceptibility during successive episodes of recurrent vulvovaginal candidiasis.
Comparative evaluation of immunoglobulin M neutralizing antibody response in acute-phase sera and virus isolation for the routine diagnosis of enterovirus infection
Authors: Pozzetto B , Gaudin O G , Aouni M , Ros A
Published: January 2020

A total of 314 patients exhibiting symptoms consistent with a viral disease provided, during the early stage of hospitalization, at least one specimen from a peripheral site (throat or stools or both) and a serum specimen in order to evaluate the neutralizing immunoglobulin M (IgM) antibody response in acute-phase serum in comparison with virus isolation for the rapid diagnosis of enterovirus (EV) infection. IgM antibodies were fractionated by ion-exchange chromatography and tested by seroneutralization against the various types of EV that have been recently circulating. A total of 189 patients (60%) were negative, and 21 (7%) were positive by both methods; in 51 patients (16%), a virus was isolated without IgM antibody response; 53 patients (17%) showed the opposite pattern. In all age groups except for children under 6 months, the frequency of positive results was higher with IgM serology than with virus isolation (27 and 22%, respectively). Apart from meningitis, for which isolation was more efficient, the other clinical conditions were associated with similar percentages of positivity by both methods. Regarding the 21 cases with positive results by the two techniques, the same serotype was detected in 9 cases and different serotypes were detected in 12, suggesting crossreactivities. Thus, IgM neutralizing antibody response on acute-phase serum appears to be of limited value in the rapid diagnosis of acute EV infection but may prove useful for the investigation of the wide range of chronic diseases associated with EV.
Antigenic and genetic variation in Legionella pneumophila serogroup 6
Authors: McKinney R M , Kuffner T A , Bibb W F , Nokkaew C , Wells D E , Arnow P M , Woods T C , Plikaytis B D
Published: January 2020

Legionella pneumophila subsp. pneumophila serogroup 6 is second in importance only to L. pneumophila serogroup 1 as a cause of legionellosis. Monoclonal antibody (MAb) reactivity and multilocus enzyme electrophoretic analyses were used to subtype serogroup 6 isolates as a potential aid for epidemiologic and virulence studies. Forty-eight serogroup 6 isolates submitted to the Centers for Disease Control from 1980 to 1985 were examined by these methods. The isolates were divided into two groups based on differential reactivity with two MAbs. Thirty-two of the isolates were of a single electrophoretic type (ET) and were reactive with both MAbs. The remaining 16 isolates were distributed among 10 ETs and were reactive with one or both MAbs. The mean genetic diversity for serogroup 6, as determined from the degree of variability at 20 enzyme loci, was found to be essentially the same as that for L. pneumophila subsp. pneumophila as a whole. The ETs of serogroup 6 isolates were unique but closely related genetically to the ETs of L. pneumophila subsp. pneumophila serogroups 1 to 5, 7, and 8. The range of serogroup 6 subtypes distinguished by MAbs and enzyme electrophoresis suggests that the combination of these two methods can be useful as a typing system.
Relationship between cytotoxic necrotizing factor production and serotype in hemolytic Escherichia coli
Authors: Caprioli A , Falbo V , Ruggeri F M , Minelli F , Orskov I , Donelli G
Published: January 2020

We examined the relationship between serotype and cytotoxic necrotizing factor (CNF) production in 123 hemolytic strains of Escherichia coli isolated from both stools and extraintestinal infections. Of 76 strains producing both hemolysin (Hly) and CNF, 66 (87%) belonged to one of six serogroups (O2, O4, O6, O22, O75, and O83). In contrast, 47 E. coli strains producing Hly only belonged to 21 different O serogroups, and only 2 of these (O6 and O18ac) were widely represented. Generally, CNF-positive and CNF-negative hemolytic isolates were assigned to different O serogroups, with the exception of O6, often present in both categories of isolates. Serogroups O4 and O18ac were significantly more prevalent among strains from extraintestinal infections than among those from stools. In contrast, the Hly-positive, CNF-negative isolates, belonging to numerous less common serogroups, were hardly ever isolated from extraintestinal infections. Serological typing further confirmed that hemolytic isolates of E. coli may grossly be divided into two main populations on the basis of the ability to produce CNF. Examination of hemolytic E. coli for this property may also be useful in achieving a more detailed characterization of pathogenic clones.
Detection of piluslike structures on clinical and environmental isolates of Vibrio vulnificus
Authors: Gander R M , LaRocco M T
Published: January 2020

Twenty clinical isolates of Vibrio vulnificus were compared with 10 environmental strains by using electron microscopy and agglutination assays with human erythrocytes, guinea pig erythrocytes, and Saccharomyces cerevisiae. In addition, the isolates were tested for ability to adhere to the human epithelial cell lines HEp-2 and A549. When examined by electron microscopy, 16 (80%) of the 20 clinical isolates demonstrated the presence of piluslike structures; the composition of the bacterial populations ranged from 0 to 68% piliated cells. In contrast, only 3 (30%) of the 10 environmental isolates were piliated, with a range from 0 to 16% piliated cells. A significant association between the presence of piliated cells and the isolate source was found (P less than 0.05). None of the 30 strains agglutinated erythrocytes or yeast cells. V. vulnificus adherence results obtained with HEp-2 cells showed 10 (50%) of 20 clinical isolates and 0 (0%) of 10 environmental isolates with averages of greater than 10 adherent bacteria per cell, demonstrating a correlation between attachment and the isolate source (P less than 0.05). Selected strains were tested to determine whether methyl alpha-D-mannopyranoside, fructose, or alpha-L-(-)-fucose would inhibit bacterial adherence to HEp-2 cells. Multiple patterns of adherence inhibition were observed. Adherence to A549 cells showed 8 (40%) of 20 clinical isolates and 0 (0%) of 10 environmental strains with averages of greater than 10 adherent bacteria per cell. A statistical association between attachment and the isolate source was demonstrated (P less than 0.05). These data suggest that the presence of piluslike structures and the ability to adhere to human epithelial cell lines may be more closely associated with V. vulnificus isolates from clinical specimens than with environmental strains.
New genome types of adenovirus types 1, 3, and 5 isolated from stools of children in Brazil
Authors: Gomes S A , Candeias J A , Monteiro S P , Pereira H G , Niel C
Published: January 2020

During an epidemiological survey made in São Paulo (Brazil), fecal specimens were periodically collected from 100 randomly chosen babies from birth to the age of 18 months. The stools, routinely collected each month and also collected each time a child presented any sign of disease, were screened for the presence of adenoviruses. Sixteen adenovirus strains, isolated from the stools of healthy and ill children, were characterized by restriction enzyme analysis. Five isolates were from subgenus A, five were from subgenus B, four were from subgenus C, and two were from subgenus D. All but two showed some restriction patterns different from those of the 42 human adenovirus prototypes and all the genome types described up to now. No fastidious adenovirus (subgenus F, serotypes 40 and 41) was encountered in the stools examined. We report here the restriction enzyme analysis of isolates of subgenera B and C. The following new designation genome types are proposed: Ad3e1 (subgenus B) and Ad1d, Ad5a1, and Ad5a2 (subgenus C).
Comparative evaluation of the Roche Cobas IDA and Enterotube II systems for identifying members of the family Enterobacteriaceae
Authors: Holmes B
Published: January 2020

Two Hoffmann-La Roche products were evaluated in parallel for their ability to identify 321 strains of the family Enterobacteriaceae. The first product was the well-established Enterotube II, which correctly identified 90% of the isolates after 24 h of incubation. The second was the ID-E rotor, originally designed for use in the Cobas Bact but here used in the new Cobas IDA system. The Cobas IDA, used with the full data base held in a microcomputer supplied with the product, identified 87% of strains after 4 h of incubation.
Identification of Campylobacter jejuni and C. coli by gel electrophoresis of the outer membrane proteins
Authors: Derclaye I , Delor I , Van Bouchaute M , Moureau P , Wauters G , Cornelis G R
Published: January 2020

Analysis of the electrophoretic profiles of the outer membrane proteins could be used to differentiate Campylobacter jejuni (16 strains) from Campylobacter coli (10 strains). This observation was confirmed by the study of DNA homology obtained by a quantitative filter hybridization method. The hippurate hydrolysis test gave a poor correlation with the results of differentiation obtained by DNA homology studies and outer membrane protein profile.
Characterization of Sicilian strains of spotted fever group rickettsiae by using monoclonal antibodies
Authors: Vitale G , Di Stefano R , Damiani G , Mansueto S
Published: January 2020

Twenty-two hybridomas producing anti-Rickettsia conorii monoclonal antibodies were obtained by nine fusion experiments. The strain chosen for immunization of mice was MAVI, an R. conorii strain isolated from a Sicilian patient with Boutonneuse fever. When tested for immunoglobulin isotype by an indirect immunofluorescence (IIF) assay, 46.6% of supernatants from the 22 hybridomas were immunoglobulin M. The supernatants were tested in the IIF assay for binding to the MAVI strain and four spotted fever group rickettsia strains isolated from Sicilian ticks (two virulent and two nonpathogenic when inoculated intraperitoneally in male guinea pigs). Only five of the supernatants showed a positive IIF result on all tested strains, although they produced different titers to the various strains, possibly an indication that they recognized an antigen common to spotted fever group rickettsiae. Immunodominant epitopes for humans were determined by using patient sera to analyze inhibition of binding to the MAVI strain. Although a limited number of serum samples were screened, a high percentage of Boutonneuse fever patients produced antibodies recognizing the same epitopes as were recognized by the mouse monoclonal antibodies. A striking heterogeneity was found both in the expression of mouse-recognized epitopes on the five rickettsial strains and in the serum antibody responses of Boutonneuse fever patients to these epitopes.
Comparison of Diamond's medium modified and Kupferberg medium for detection of Trichomonas vaginalis
Authors: Gelbart S M , Thomason J L , Osypowski P J , James J A , Hamilton P R
Published: January 2020

Diamond's medium modified, the commercially available version of the Klaas modification of Diamond's medium, was compared to Kupferberg medium and to direct wet-mount examination for detection of trichomonads in symptomatic patients. Diamond's medium was found to be superior to both Kupferberg medium (P less than 10(-4) and wet mount (P less than 10(-6].
Novel phase-shift marker in cell surface proteins of Bordetella bronchiseptica
Authors: Kuzuya M , Kodama Y
Published: January 2020

Cell surface proteins of phase I cultures of Bordetella bronchiseptica strains from various species of animals were compared with those of isogenic phase III cultures by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A protein band with a molecular mass of 74 kilodaltons was found only in various phase I cultures. This protein disappeared in parallel with antigenic modulation and had strong antigenicity. We found it to be an appropriate phase-shift marker for two reasons: it was readily extracted and recognized, and it was specifically identified by its distinct antigenicity.
B.CAT CONFIRM, a rapid test for confirmation of Branhamella catarrhalis
Authors: Janda W M , Ruther P
Published: January 2020

B.CAT CONFIRM (Scott Laboratories, Inc., Fiskeville, R.I.), a rapid test for detection of tributyrin hydrolysis, was evaluated for its ability to identify strains of Branhamella catarrhalis and to differentiate them from Neisseria species and related species. On initial testing, B.CAT CONFIRM was positive for 65 (96%) of 68 B. catarrhalis strains within 30 min after inoculation. Retesting of the remaining three strains resulted in their correct identification. B.CAT CONFIRM was negative for all Neisseria spp. (130 strains) and for Kingella spp. (3 strains). Two of the three Moraxella spp. were weakly positive in the B.CAT CONFIRM after 60 min, but these reactions were easily distinguishable from the strong reactions of B. catarrhalis strains. This test will be helpful in the clinical laboratory for the rapid identification of B. catarrhalis in clinical specimens.
Microbiology of postthoractomy sternal wound infection
Authors: Brook I
Published: January 2020

Specimens from 74 patients with postthoractomy sternal wound infection were studied for aerobic and anaerobic bacteria. Bacterial growth was obtained in specimens from 65 patients (88%). Aerobic or facultative bacteria only were recovered in 50 specimens (77%), anaerobic bacteria only in 6 (9%), and mixed aerobic, facultative, and anaerobic bacteria in 9 (14%). Eighty-seven isolates were recovered (1.3 per specimen): 68 aerobic or facultative (1.0 per specimen) and 19 anaerobic (0.3 per specimen). The predominant aerobes were Staphylococcus epidermidis (28 isolates), Staphylococcus aureus (21 isolates), and members of the family Enterobacteriaceae (14 isolates). The predominant anaerobes were Peptostreptococcus spp. (10 isolates), Bacteroides spp. (4 isolates), and Clostridium spp. (3 isolates). Polymicrobial infection occurred in 18 instances (28%). A single organisms was recovered in 47 instances (72%); these included 20 isolates of S. epidermidis, 15 of S. aureus, 5 of Enterobacteriaceae, and 4 of anaerobes. These data highlight the previously unrecognized polymicrobial aerobic and anaerobic bacteriology in a percentage of patients with postthoractomy sternal wound infection.
Wound infection caused by Branhamella catarrhalis
Authors: Gray L D , Van Scoy R E , Anhalt J P , Yu P K
Published: January 2020

Branhamella catarrhalis was isolated from sputum, tracheal secretions, and a nonhealing and infected thoracic surgical wound in a 59-year-old woman who had a history of a chronic, interstitial fibrosis and who had undergone an open lung biopsy procedure. The patient's upper respiratory tract was the likely source of the organism. To our knowledge, this is the first report of a wound infection caused by B. catarrhalis.
Levels of antibody to defined antigens of Mycobacterium tuberculosis in tuberculous meningitis
Authors: Chandramuki A , Bothamley G H , Brennan P J , Ivanyi J
Published: January 2020

The aim of the study was to develop an antibody immunoassay for tuberculous meningitis with a sensitivity greater than that of the initial bacterial smear while maintaining a 100% specificity. Antibody titers to five purified antigens of Mycobacterium tuberculosis were measured in cerebrospinal fluid by using an enzyme-linked immunosorbent assay technique. Seventy-four patients with tuberculous meningitis (26 culture positive) were compared with 26 patients with purulent meningitis, 69 patients with suspected but excluded tuberculous meningitis, and 29 patients with other neurological diseases. Antibody responses to both the 14-kilodalton (kDa) antigen and lipoarabinomannan were immunodominant, accounting for 74% of those with any antibody titer to a soluble extract of M. tuberculosis. A sensitivity of 61% was achieved (77% for the culture-positive samples) by using immunoglobulin G titers to lipoarabinomannan and the 14-, 19-, and 38-kDa antigens. Thus, estimation of levels of antibody in cerebrospinal fluid to selected mycobacterial antigens would be valuable in the diagnosis of tuberculous meningitis. The possible reasons for the immunodominance of the 14-kDa antigen and lipoarabinomannan in the pathogenesis of tuberculous meningitis are discussed.
Four new provisional serovars of Shigella
Authors: Gross R J , Thomas L V , Cheasty T , Rowe B , Lindberg A A
Published: January 2020

Four bacterial strains are described that possess the biochemical characteristics of Shigella species but do not belong to any of the established Shigella serovars or to any previously described provisional serovar. One strain fermented mannitol, and it is proposed that this be the type strain for a new provisional serovar of Shigella boydii. The remaining strains did not ferment mannitol and belonged to three different serovars. These strains are proposed as type strains for three new provisional serovars of Shigella dysenteriae. All four strains were invasive in a HEp-2 cell tissue culture test, but only one was invasive in the guinea pig eye test and might therefore be expected to cause dysenterylike illness in humans. It is important that the designation of such strains remain provisional until other reference laboratories have had the opportunity to search for additional isolates and the possible pathogenicity of these strains for humans can be further assessed.
Purification of a Brucella canis cell wall antigen by using immunosorbent columns and use of the antigen in enzyme-linked immunosorbent assay for specific diagnosis of canine brucellosis
Authors: Serikawa T , Iwaki S , Mori M , Muraguchi T , Yamada J
Published: January 2020

A cell wall antigen of Brucella canis was purified by immunosorbent columns. The antigen contained two proteins of 30 and 28 kilodaltons and a polysaccharide exhibiting a 12-kilodalton band upon 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antibody to the purified antigen, which specifically reacted with the polysaccharide, was used as the first coating antibody in an enzyme-linked immunosorbent assay (ELISA) for serological diagnosis of canine brucellosis. Dogs inoculated orally with live B. canis were positive and dogs from B. canis-free colonies were negative in the ELISA. Of 199 dogs from a brucellosis-contaminated area, 116 with negative titers in the tube agglutination test (TAT), using heat-inactivated whole B. canis cells as the antigen, were also negative in the ELISA. Seventy-eight of the dogs with questionable titers in the TAT were divided into two groups: 20 dogs that were positive in the ELISA and 58 that were negative. Of five dogs with positive titers in the TAT, three were positive in the ELISA and the gel immunodiffusion test (GD) with crude B. canis extract as the antigen and were also culture positive for B. canis. One dog was positive in the ELISA and GD but gave a negative culture result. Serum from the remaining dog, which was positive with high titer in the TAT but negative in the ELISA and in culture for B. canis, formed a spur precipitate with a homologous precipitate in the GD. These results indicate that the ELISA is a specific serological test for B. canis infection in dogs.
Monitoring human immunodeficiency virus type 1-infected patients by ratio of antibodies to gp41 and p24
Authors: Schmidt G , Amiraian K , Frey H , Wethers J , Stevens R W , Berns D S
Published: January 2020

Antibody responses of 85 patients to human immunodeficiency virus type 1 antigens were quantitated by densitometric analysis of Western blot (immunoblot) assays. All patients had been classified into the following three clinical categories: asymptomatic (ASY), acquired immunodeficiency syndrome (AIDS)-related complex (ARC), or AIDS. Fifty of the patients were monitored for 6 to 29 months. The gp41/p24 antibody ratio was examined in three studies. In the first study, initial specimens from each patient were analyzed. The mean gp41/p24 antibody ratios were 1.5 (ASY), 3.2 (ARC), and 5.4 (AIDS). Of ASY patients, 79% had antibody ratios of less than 2.0. In contrast, 72% of patients with AIDS had ratios of greater than or equal to 2.0. In the second study, serially obtained specimens from ASY, ARC, and AIDS patients were analyzed. These patients were further grouped according to progression of their clinical condition. Of ASY patients whose clinical condition progressed to ARC, 80% consistently had ratios of greater than or equal to 2.0. Of ARC patients whose clinical condition progressed to AIDS, 71% consistently had ratios of greater than or equal to 2.0. Of AIDS patients who died during the study, 100% consistently had ratios of greater than or equal to 2.0. No patients were treated with azidothymidine during the first two studies. In the third study, AIDS patients were monitored before and during treatment with azidothymidine. During treatment, ratios stabilized or improved transiently in five of seven patients. In these three studies, a gp41/p24 antibody ratio of less than 2.0 correlated with a benign clinical state and a ratio of greater than or equal to 2.0 correlated with AIDS or progression to AIDS.
Enzyme polymorphism, prodigiosin production, and plasmid fingerprints in clinical and naturally occurring isolates of Serratia marcescens
Authors: Gargallo-Viola D
Published: January 2020

Enzyme polymorphism and genetic relationship among 99 Serratia marcescens isolates obtained from clinical and environmental sources were determined by analysis of electromorphs in nine enzyme loci encoded by chromosomal genes. Seven of the loci were polymorphic, and 33 distinctive electrophoretic types (ETs) representing multilocus genotypes were identified. Cluster analysis, based on the proportion of mismatches between multilocus genotypes, revealed two clearly differentiated groups of ETs in S. marcescens. One was represented exclusively by isolates with nonchromogenic biotypes recovered almost entirely (97.3%) from clinical samples. The other group comprised all isolates characterized by the production of prodigiosin or by belonging to a chromogenic biotype. Absolute correlation was found between the ability to produce prodigiosin and the absence of plasmids. In contrast, 24% of the nonchromogenic isolates contained plasmids. Results obtained by analysis of multilocus genotypes were related to those obtained by biotyping and plasmid fingerprinting. However, more groups could be distinguished by analysis of ETs than by biotyping. Plasmid fingerprinting was a limited typing system because many isolates lacked plasmids. Although the results of this study did not permit a definitive correlation between ETs and pathogenicity of the isolates, more detailed studies of these groups will help to understand the different clinical significances of the nonchromogenic and chromogenic isolates of S. marcescens.
In situ hybridization for quantitative assay of infectious hepatitis A virus
Authors: Xi J N , Estes M K , Metcalf T G
Published: January 2020

A method of in situ hybridization using single-stranded RNA probes of opposite polarity for quantitative enumeration of hepatitis A virus (HAV) in infected cells has been developed. Kinetic experiments showed that foci of infected cells appeared as early as day 2 postinfection. The absence of foci in cells examined immediately after virus adsorption indicated that foci detected subsequently were related to viral replication. Foci were detected by hybridization with RNA probes complementary to HAV genomic RNA but not with RNA probes identical to HAV genomic RNA. The number of foci observed was linearly related to the HAV dose inoculated. Focus formation was reduced when a virus inoculum was pretreated with guinea pig anti-HAV hyperimmune serum but not when it was pretreated with preimmune serum. The high resolution of hybridization signals and relative rapidity of the test indicated that this technique will be useful for measuring serum neutralizing antibodies and for quantitative assay of infectious HAV.
Evaluation of a latex agglutination test for Clostridium difficile in two nursing home outbreaks
Authors: Bennett R G , Laughon B E , Mundy L M , Bobo L D , Gaydos C A , Greenough W B , Bartlett J G
Published: January 2020

The Culturette Brand Clostridium difficile test (CDT; Marion Laboratories, Inc., Kansas City, Mo.) is a latex agglutination test for C. difficile. The recent controversy involving the identity of antigens detected by CDT has made decisions on its use difficult. We compared the test results with those of selective culture and stool cytotoxin assays in investigations of two nursing home outbreaks of C. difficile-associated disease in order to formulate usage recommendations. Selective culture for C. difficile identified 27 (19%) of 142 subjects as carriers. CDT and the stool cytotoxin assay identified only 52 and 48% of these carriers, respectively. Compared with the stool cytotoxin assay, CDT had a high sensitivity (92%) and specificity (89%) for the detection of C. difficile disease, but the positive predictive value of the test was only 17% when the prevalence of disease was 2%. We conclude that the CDT should not be used to identify carriers but that it is a sufficiently sensitive and specific screening test for diagnosing C. difficile disease. However, since the positive predictive value of the CDT is low when the prevalence of disease is low, positive test results should be confirmed by the stool cytotoxin assay.
Effect of differences in specimen processing and passage technique on recovery of Chlamydia trachomatis
Authors: Jones R B , Van Der Pol B , Katz B P
Published: January 2020

We have previously found that optimum recovery of Chlamydia trachomatis in microdilution plate culture required multiple blind passages. However, others have found this not to be the case for culture in vials. In the present study, the effect on recovery of the use of vials (as opposed to microdilution plates) and the effect of vortexing, sonication, or both were compared. Three different passage techniques were also evaluated. Vortexing or sonication resulted in equivalent recoveries. However, compared with vortexing alone, a combination of vortexing and sonication increased recovery from 95 (78%) to 114 (94%) of 121 positive specimens (P = 0.002). In multiple-passage experiments, the combination of vortexing and sonication, compared with vortexing only, increased the proportion of isolates recovered with no more than a single passage from 81 to 96%. Substitution of vials for microdilution plates increased recovery with only a single passage to greater than 96%, irrespective of whether sonication was employed. The most sensitive technique for single-passage technique was one using blunt scraping of cell monolayers with passage of two monolayers to one. The sensitivity of cell culture for C. trachomatis is highly dependent on the technique(s) employed. However, the combination of sonication and vortexing of clinical specimens enhanced recovery in microdilution plates, and a single blind passage did so in both microdilution plates and vials. Consideration should be given to their use for routine clinical cultures.
Phenotypic characteristics of Branhamella catarrhalis strains
Authors: Soto-Hernandez J L , Holtsclaw-Berk S , Harvill L M , Berk S L
Published: January 2020

Isolates of Branhamella catarrhalis from 13 patients with pneumonia, 6 patients with tracheobronchitis, and 8 patients who were colonized with the organism were studied with respect to susceptibility to the bactericidal action of normal human serum (NHS), glass slide hemagglutination (HA) of group O human erythrocytes, beta-lactamase production, and susceptibility to selected antimicrobial agents and laboratory drugs. A total of 18 of 27 isolates were serum resistant, 22 of 27 produced HA, and 21 of 27 were beta-lactamase positive. Statistically significant correlations were found between susceptibility to NHS and susceptibility to trypsin (r = +0.47; P = 0.01) and between susceptibility to NHS and HA (r = -0.48; P = 0.009). Significant correlations were also observed among several pairs of antimicrobial drugs. Analysis of variance showed that mean ampicillin MICs correlated with isolate group (r = -0.49; P = 0.03) in that the pneumonia isolates had higher MICs. Some phenotypic characteristics appeared to be independent of each other. These data suggest that important differences exist among clinically significant B. catarrhalis strains and that these differences may be due to differences in the cell wall envelope of the organism.
Evaluation of cold-recombinant influenza A/Korea (CR-59) virus vaccine in infants
Authors: Anderson E L , Belshe R B , Burk B , Bartram J , Maassab H F
Published: January 2020

Twenty-four infants 5 to 13 months of age were intranasally vaccinated with a live cold-recombinant influenza A/Korea (CR-59, H3N2) virus vaccine. Nineteen infants served as controls. The inocula ranged from 10(3.2) to 10(6.2) 50% tissue culture infective doses (TCID50) per infant. Zero of six, one of four, seven of ten, and four of four infants receiving 10(3.2), 10(4.2), 10(5.2), and 10(6.2) TCID50, respectively, were infected by the intranasal vaccine. The amount of virus required to infect 50% of infants was calculated to be 10(4.6) TCID50. The occurrence of fever, respiratory illness, and otitis media was common among both controls and vaccinees in the postinoculation period. Maternal antibody was present in low titers in some infants and did not inhibit replication of the vaccine virus.
Bacteriological and electron microscopy examination of brown pigment stones
Authors: Leung J W , Sung J Y , Costerton J W
Published: January 2020

Bacteriological and morphological studies of 38 brown pigment common duct stones were performed. Stone cultures were positive for bacteria in 80.5% of those studied. Enterococci were the most common organisms that were isolated. Scanning electron microscopy showed the presence of bacteria in 84.2% of the stones. The bacteria were seen embedded within an amorphous matrix in alternating layers of flakelike crystals. Transmission electron microscopy showed the presence of gram-positive and gram-negative bacteria surrounded by a ruthenium red-stained exopolysaccharide material. Results of the bacteriological and morphological studies confirmed the close relationship between the presence of bacteria and the development of brown pigment stones.
Investigation of atypical western blot (immunoblot) reactivity involving core proteins of human immunodeficiency virus type
Authors: Josephson S L , Swack N S , Ramirez M T , Hausler W J
Published: January 2020

Serum specimens which originally exhibited a narrow (indeterminate) 24-kilodalton core protein (p24) or p24/p55 pattern of reactivity with human immunodeficiency virus (HIV) in the Western blot (immunoblot) test were studied to gather information on antibody specificity. A total of 12 specimens were initially reevaluated with an indirect immunofluorescence assay (IFA), three enzyme-linked immunosorbent assays (ELISAs), and Western blot analyses. Five of the specimens were IFA positive and contained anti-gp160/gp120 antibodies which were observed only when an HIV Western blot antigen rich in gp160 and gp120 was used. The remaining seven serum specimens were nonreactive by IFA and showed variable reactivity in HIV antibody ELISAs. The specimens did not cross-react with core antigens for human T-cell leukemia virus types 1 and 2 or contain detectable levels of HIV p24 antigen. The p24/p55 reactivity of six of the seven indeterminate specimens could be reduced or eliminated by preincubating the specimens with disrupted, HIV-infected H9 cells but not with uninfected H9 cells. The six specimens also exhibited discernible reactivity with recombinant HIV p24 antigen. When an additional 23 indeterminate specimens were assayed, all of the serum specimens were nonreactive by IFA while 65% (15 of 23) showed various degrees of reactivity with the recombinant p24 protein. There was no indication that any of the HIV core antibody reactivity was caused by HIV infection. Indeterminate results for five patients with specific p24 reactivity, who were retested after a period of weeks or months, remained indeterminate for HIV antibody with no significant change in ELISA or Western blot reactivity.
Cellular fatty acid composition of Campylobacter pylori from primates and ferrets compared with those of other campylobacters
Authors: Goodwin C S , McConnell W , McCulloch R K , McCullough C , Hill R , Bronsdon M A , Kasper G
Published: January 2020

The cellular fatty acid profiles of newly described campylobacters were determined on a polar, capillary column. Six isolates of the gastric spiral organism, Campylobacter pylori subsp. mustelae, from ferrets from Australia, England, and the United States were all found to have a similar fatty acid profile which was different from that of C. pylori from humans; C. pylori subsp. mustelae did not have 3-hydroxyoctadecanoic acid (3-OH C18:0) and had much less tetradecanoic acid (C14:0) and much more hexadecanoic acid (C16:0). Inasmuch as Lambert et al. (M.A. Lambert, C.M. Patton, T.J. Barrett, and C.W. Moss, J. Clin. Microbiol. 25:706-713, 1987) have proposed that campylobacters can be grouped by cellular fatty acid composition, we propose this organism should be in a new gas-liquid chromatography (GLC) group, group J. Seven isolates of gastric spiral organisms from macaque monkeys and baboons, including three from Macaca nemestrina, and one isolate from a pig were found to have fatty acid profiles very similar to that of C. pylori; but a second type of organism (type B) from M. nemestrina had a unique profile without 19-carbon cyclopropane fatty acid (C19:0 cyc) but with 3-hydroxy tetradecanoic acid (OH C14:0), which is not present in other gastric spiral bacteria. We propose that this organism (nemestrina type B) should be in a new GLC group, group K. The cellular fatty acid profile of seven isolates of C. jejuni subsp. doylei was found to be similar to that for C. jejuni, but with possibly significant differences in that the former did not have 3-OH C14:0 but did have 3-hydroxyhexadecanoic acid (3-OH C16:0) and had more C14:0 than did C. jejuni. Two strains of urease-positive thermophilic campylobacters were found to have a profile similar to that of "C. cinaedi" and thus should be included with them in GLC group D. We confirm that C. sputorum has a unique cellular fatty acid composition and suggest that it should be in a new group, group H.
Rapid diagnosis of acute Epstein-Barr virus infection by an indirect enzyme-linked immunosorbent assay for specific immunoglobulin M (IgM) antibody without rheumatoid factor and specific IgG interference
Authors: Ho D W , Field P R , Cunningham A L
Published: January 2020

An indirect enzyme-linked immunosorbent assay (ELISA) for detection of Epstein-Barr virus-specific immunoglobulin M (IgM) antibody was developed with commercial reagents. Sera containing rheumatoid factor (RF) (as little as 0.5 IU/ml) coupled with specific IgG resulted in false-positives in the ELISA. This interference was eliminated by the use of anti-human IgG antibodies to remove RF and IgG. Thus, pathogen-specific IgG complexes to which IgM-RF could be bound during the subsequent test were inhibited, and competition between specific IgG and IgM was also prevented. Of the 1,672 serum specimens tested, 353 were found to be Epstein-Barr virus IgM antibody positive by indirect immunofluorescence (IF). Compared with the IF test, the ELISA showed 96.6% sensitivity, 99.7% specificity, and 99% accuracy. Further evidence indicated that most of the 12 ELISA false-negatives were IF false-positives. There was a linear correlation between mean ELISA values and increasing IF titers (r = 0.96). However, the IF test has the disadvantages that it lacks automated reading and requires considerable technical expertise, both of which restrict the range of laboratories performing the test. The indirect ELISA has the advantages that it is simple and rapid and can be automated. All the reagents used in this assay are commercially available, have been prestandardized, and are stable.
Cleavage of procaryotically expressed human immunodeficiency virus fusion proteins by factor Xa and application in western blot (immunoblot) assays
Authors: Ellinger S , Glockshuber R , Jahn G , Plückthun A
Published: January 2020

The proteins p15 and p24 of the human immunodeficiency virus (HIV) type 1 gag gene were expressed as fusion proteins in Escherichia coli for detecting antibodies against the acquired immunodeficiency virus by Western blot (immunoblot) analysis. These fusion proteins contain amino acids 1 to 375 of the E. coli beta-galactosidase linked to the viral protein(s) by a recognition sequence for the specific protease factor Xa. They are obtained in large amounts in insoluble inclusion bodies. To avoid ambiguous results caused by cross-reaction of sera with bacterial proteins in Western blots, we purified the recombinant fusion proteins and subsequently removed the bacterial part of the fusions by cleavage with factor Xa. The cleavage mixtures were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted onto nitrocellulose membranes. The viral proteins obtained by this method did not contain any bacterial proteins or protein fragments. Thus, false-positive results in HIV Western blot analysis with bacterially expressed HIV proteins can be excluded with these purified recombinant viral antigens.