bluuebunny/crossref_metadata_2025 huggingface dataset:
Prospective multicenter study of vascular-catheter-related complications and risk factors for positive central-catheter cultures in intensive care unit patients
Authors: Richet H , Hubert B , Nitemberg G , Andremont A , Buu-Hoi A , Ourbak P , Galicier C , Veron M , Boisivon A , Bouvier A M
Published: January 2020

To determine the incidence rate of complications associated with vascular catheters in intensive care unit patients and to analyze risk factors for a positive vascular culture, we performed a multicenter study of intensive care unit patients at eight French hospitals. During the study period, 865 intravenous catheters were inserted in 566 patients; 362 (41.8%) were peripheral catheters, and 503 (58.2%) were central catheters. Local complications (i.e., infiltration) occurred significantly more often with peripheral than with central catheters (P less than 0.001); in contrast, fever and bacteremia were significantly more often associated with central than with peripheral catheters (P less than 0.01 and P less than 0.05, respectively). The culture of the vascular-catheter tip was positive for 24% of central catheters (32 of 1,000 catheters days) and for 9% of peripheral catheters (21 of 1,000 catheters days). Staphylococcus epidermidis was the most common microorganism isolated from both peripheral and central catheters, followed by Staphylococcus aureus and Pseudomonas aeruginosa. No significant risk factor associated with positive cultures for peripheral catheters was found by univariate analysis. In contrast, the purpose of the cannula (nutrition and monitoring of central venous pressure), the insertion site (jugular), the dressing type (semipermeable transparent dressing), the antiseptic used to prepare the insertion site (povidone iodine), and routine changing of the intravenous administration set were significantly associated with positive cultures of central catheters. Three factors, duration of catheterization, use of a semipermeable transparent dressing, and the jugular insertion site, were found to be independently associated with positive cultures of central catheters by multivariate analysis.
Serotypic differentiation of group A rotaviruses with porcine rotavirus gene 9 probes
Authors: Rosen B I , Saif L J , Jackwood D J , Gorziglia M
Published: January 2020

The serotypic specificities of Gottfried and OSU porcine rotavirus gene 9 probes were investigated in a dot hybridization assay. The probes were reacted with homologous and heterologous serotypes of group A rotaviruses of human and animal origin. Hybridizations were conducted under relatively low-stringency (52 degrees C, no formamide, 5 x SSC) and high-stringency (52 degrees C, 50% formamide, formamide, 5 x SSC) conditions (1 x SSC is 0.15 M NaCl plus 0.015 M sodium citrate). Under conditions of relatively low stringency, the Gottfried and OSU gene 9 probes demonstrated broad cross-reactivity and were useful in the detection of homologous and heterologous serotypes of group A rotaviruses. Under conditions of relatively high stringency, the Gottfried and OSU gene 9 probes were serotype specific. The Gottfried gene 9 probe (serotype 4) hybridized with homologous Gottfried porcine rotavirus as well as the serotype 4 human rotaviruses ST3 and VA70. The OSU gene 9 probe (serotype 5) hybridized with homologous OSU porcine rotavirus and the serotype 5 equine rotavirus H1. Hybridization was not observed with the antigenically distinct group B and C porcine rotaviruses or with other porcine enteric viruses, including calicivirus and a coronavirus, transmissible gastroenteritis virus, regardless of stringency conditions. Analysis of 14 group A rotavirus-positive field samples resulted in the serotypic differentiation, collectively, of six serotype 4 or 5 porcine rotaviruses. No field samples reacted with both the Gottfried and OSU gene 9 probes.
Enhancement of anti-influenza A virus cytotoxicity following influenza A virus vaccination in older, chronically ill adults
Authors: Gorse G J , Belshe R B
Published: January 2020

We studied anti-influenza cytotoxicity by bulk peripheral blood mononuclear leukocyte (PBL) cultures derived from older, chronically ill volunteers undergoing vaccination. Vaccinees received either cold-recombinant, live-attenuated influenza A/Korea/1/82 (H3N2) virus intranasally or inactivated monovalent influenza A/Taiwan/1/86 (H1N1) subvirion vaccine intramuscularly. PBL were collected pre- and postvaccination and in vitro stimulated by autologous PBL infected with influenza A virus homologous and heterosubtypic to the respective vaccine strain. Cytotoxicity was measured against influenza A virus-infected autologous and human leukocyte antigen (HLA)-mismatched PBL targets infected with influenza A virus homologous or heterosubtypic to the vaccine virus strain. Vaccinees infected with the live-attenuated virus developed significant rises in mean anti-influenza, HLA-restricted cytotoxicity that was cross-reactive against influenza A viruses homologous and heterosubtypic to the vaccine virus. The enhanced cross-reactive cytotoxicity was inducible postvaccination by in vitro stimulation with autologous PBL infected with the homologous influenza A (H3N2) virus and with influenza A (H1N1) virus. In contrast, after vaccination with inactivated monovalent subvirion vaccine, volunteers developed significant increases in mean anti-influenza, HLA-restricted cytotoxicity only against autologous PBL infected with homologous influenza A (H1N1) virus. Increased cytotoxicity occurred only after in vitro stimulation with autologous cells infected with homologous influenza A (H1N1) virus. Mean gamma interferon levels in supernatant fluids of influenza A virus-stimulated effector PBL did not increase postvaccination, despite increased levels of anti-influenza cytotoxicity displayed by the effector cells. We conclude that the live-attenuated influenza A virus infection induced a broader range of enhanced anti-influenza cytotoxicity than did the inactivated subvirion vaccine.
Assessment of a new hub design and the semiquantitative catheter culture method using an in vivo experimental model of catheter sepsis
Authors: Segura M , Alía C , Valverde J , Franch G , Torres Rodríguez J M , Sitges-Serra A
Published: January 2020

An in vivo model of hub-related catheter sepsis in rabbits is reported. The model was used to investigate the protection offered by a new hub design against external contamination by Pseudomonas aeruginosa or Staphylococcus epidermidis and to reassess the diagnostic value of the semiquantitative culture method in bacteremia of endoluminal origin. Contamination of conventional Luer-Lock connectors was followed by clinical sepsis, positive catheter segment cultures, or both, whereas contamination of the new hub was followed by complete protection. Clinical and bacteriological discrepancies observed between contamination with P. aeruginosa and S. epidermidis suggest that the virulence of microorganisms may account for differences in the natural history of hub-originated catheter sepsis. The semiquantitative extraluminal method for catheter culture yielded less than 15 CFU in three animals with proven bacteremia and should not be used as the "gold standard" to define catheter-related bacteremia.
Neisseria elongata subsp. nitroreducens subsp. nov., formerly CDC group M-6, a gram-negative bacterium associated with endocarditis
Authors: Grant P E , Brenner D J , Steigerwalt A G , Hollis D G , Weaver R E
Published: January 2020

CDC group M-6 is the vernacular name given to a gram-negative, oxidase-positive, aerobic, nonmotile, rod-shaped bacterium. This organism is biochemically similar to Kingella denitrificans and displays a cellular fatty acid profile consistent with CDC groups M-5 and EF-4 and with Neisseria elongata. Of the 95 M-6 strains referred to the Centers for Disease Control (CDC) for identification, 32 (64%) of the first 50 were from the throat or sputum and only 3 (6%) were from blood; only 5 (11%) of the next 45 isolates were from the upper respiratory tract and 23 (51%) were from blood, with many of these (15 or 65%) being associated with endocarditis. The major characteristics of CDC group M-6 include reduction of nitrate and nitrite with no gas formation; positive reaction for oxidase; negative reactions for catalase, urease, indole, and motility; and no acid production from carbohydrates. Guanine-plus-cytosine content determined spectrophotometrically by thermal denaturation was 55 to 58 mol % for six M-6 strains tested: 56 mol % for the N. elongata subsp. elongata type strain and for the N. elongata subsp. glycolytica type strain. By the hydroxyapatite method, DNAs from 24 M-6 strains showed an average of 78% relatedness to M-6 reference strain B1019 in reactions at 60 degrees C and 73% relatedness in reactions at 75 degrees C. M-6 strain B1019 was 79% related to the N. elongata type strain at 60 degrees C and 71% related at 75 degrees C; it was 75% related to the type strain N. elongata subsp. glycolytica at 60 degrees C and was 66% related at 75 degrees C. DNAs from CDC group EF-4, K. denitrificans, and CDC group M-5 were all less than 14% related to CDC group M-6 at 75 degrees C. The DNA relatedness data showed conclusively that all the M-6 strains belong in the species N. elongata. M-6 is different from N. elongata subsp. elongata in that M-6 reduces nitrate and sometimes weakly acidifies D-glucose, and it is different from N. elongata subsp. glycolytica in that it reduces nitrate and is negative for glucose and catalase. Because of the apparent clinical significance of M-6 compared with the clinical significance of N. elongata subsp. elongata and N. elongata subsp. glycolytica and the ease in distinguishing it biochemically, we propose M-6 as a third subspecies of N.elongata, N. elongata subsp. nitroreducens subsp. nov.
Comparative analysis of human cytomegalovirus a-sequence in multiple clinical isolates by using polymerase chain reaction and restriction fragment length polymorphism assays
Authors: Zaia J A , Gallez-Hawkins G , Churchill M A , Morton-Blackshere A , Pande H , Adler S P , Schmidt G M , Forman S J
Published: January 2020

The human cytomegalovirus (HCMV) a-sequence (a-seq) is located in the joining region between the long (L) and short (S) unique sequences of the virus (L-S junction), and this hypervariable junction has been used to differentiate HCMV strains. The purpose of this study was to investigate whether there are differences among strains of human cytomegalovirus which could be characterized by polymerase chain reaction (PCR) amplification of the a-seq of HCMV DNA and to compare a PCR method of strain differentiation with conventional restriction fragment length polymorphism (RFLP) methodology by using HCMV junction probes. Laboratory strains of HCMV and viral isolates from individuals with HCMV infection were characterized by using both RFLPs and PCR. The PCR assay amplified regions in the major immediate-early gene (IE-1), the 64/65-kDa matrix phosphoprotein (pp65), and the a-seq of the L-S junction region. HCMV laboratory strains Towne, AD169, and Davis were distinguishable, in terms of size of the amplified product, when analyzed by PCR with primers specific for the a-seq but were indistinguishable by using PCR targeted to IE-1 and pp65 sequences. When this technique was applied to a characterization of isolates from individuals with HCMV infection, selected isolates could be readily distinguished. In addition, when the a-seq PCR product was analyzed with restriction enzyme digestion for the presence of specific sequences, these DNA differences were confirmed. PCR analysis across the variable a-seq of HCMV demonstrated differences among strains which were confirmed by RFLP in 38 of 40 isolates analyzed. The most informative restriction enzyme sites in the a-seq for distinguishing HCMV isolates were those of MnlI and BssHII. This indicates that the a-seq of HCMV is heterogeneous among wild strains, and PCR of the a-seq of HCMV is a practical way to characterize differences in strains of HCMV.
Antifusion activity in sera from persons infected with human immunodeficiency virus type 1
Authors: Graham B S , Rowland J M , Modliszewski A , Montefiori D C
Published: January 2020

Cell-to-cell fusion plays an important role in the pathogenesis of human immunodeficiency virus type 1 (HIV-1) infections. An assay to measure the antifusion activity of serum has been developed by using the fusion event that occurs between H9 cells chronically infected with HIV-1 (H9IIIB) and fusion-susceptible MT-2 cells. The endpoint is determined by measuring neutral red uptake in cells after syncytium formation is allowed to occur in the presence of various serum dilutions. The assessment of antifusion activity in serum by neutral red uptake has been shown to correlate with syncytium reduction as determined by direct counting. The optimal number and ratio of cells in the suspension for efficiency and speed of the assay have been determined. With this assay it was shown that 50% of 36 serum specimens capable of neutralizing cell-free virions failed to inhibit syncytium formation. The assay can thus measure a distinct activity in HIV-1-immune human sera which is a subset of neutralization activity. Because of the potential role of this activity in the rate of disease progression and protective immune responses, the antifusion assay will be an important tool for the investigation of disease pathogenesis and for acquired immunodeficiency syndrome vaccine development. The assay can also be applied to the investigation of the pathogenesis of the fusion event at the cellular level. The ability to use absorbance measurements rather than syncytium counts as the endpoint facilitates direct computer-assisted data analysis, which expedites the performance of the assay.
Expression of type 8 capsular polysaccharide and production of toxic shock syndrome toxin 1 are associated among vaginal isolates of Staphylococcus aureus
Authors: Lee J C , Liu M J , Parsonnet J , Arbeit R D
Published: January 2020

A colony immunoblot method was developed for serotyping the capsular polysaccharides expressed by Staphylococcus aureus isolates. The method was rapid and specific and was performed with either polyclonal or monoclonal antibodies specific for each of the capsule types. S. aureus isolates were obtained from patients with toxic shock syndrome (TSS) or other staphylococcal infections and from asymptomatic women with vaginal colonization. Among the vaginal isolates of S. aureus, expression of the type 8 capsule was significantly (P less than 0.001) more frequent among strains that produced TSS toxin 1 (TSST-1) than it was among TSST-1-negative strains. In contrast, the frequency of type 8 capsule expression was similar among both TSST-1-positive and -negative strains of S. aureus from patients with nonvaginal TSS. When all vaginal and nonvaginal isolates were compared, TSST-1-negative S. aureus strains were equally distributed among the type 5 and 8 and nontypeable capsule groups, whereas TSST-1-positive strains were predominantly capsule type 8.
Coaggregation of oral Candida isolates with bacteria from bone marrow transplant recipients
Authors: Hsu L Y , Minah G E , Peterson D E , Wingard J R , Merz W G , Altomonte V , Tylenda C A
Published: January 2020

In vitro coaggregation between Candida species isolated from immunosuppressed bone marrow transplant recipients and oral bacteria was investigated. Each Candida strain showed a different pattern of coaggregation with the 22 bacterial strains studied. Two strains of Lactobacillus amylovorus isolated from separate bone marrow transplant patients and Fusobacterium nucleatum (VPI 10197) coaggregated with all Candida strains. Ten bacterial strains showed no coaggregation with the Candida strains. A variety of inhibition patterns were observed when coaggregating strains were first incubated with various sugars or subjected to heat treatment. Positive and negative results were generally consistent with all Candida strains. On the basis of the culture characteristics of the oral rinse specimens, relationships between the colonization of bacteria and yeasts and in vitro coaggregation were suggested.
Modification of the silver staining technique to detect lipopolysaccharide in polyacrylamide gels
Authors: Fomsgaard A , Freudenberg M A , Galanos C
Published: January 2020

A silver staining method used routinely for detecting bacterial lipopolysaccharide (LPS) in sodium dodecyl sulfate-polyacrylamide gels (C. Tsai and E. Frasch, Anal. Biochem. 119:115-119, 1982) appeared to be inappropriate for visualizing certain LPS preparations. It did not stain S-form fractions of polyagglutinable Pseudomonas aeruginosa LPS or several partly deacylated (alkali-treated) S-form LPS after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. However, these LPS preparations could be detected by anti-LPS sera after electroblotting onto nitrocellulose, thereby confirming their integrity and presence in the polyacrylamide gel. This is because LPS fractions containing a low number of fatty acids are washed out of the gel during the initial fixing step (40% ethanol-4% acetic acid, overnight). By omitting this fixing step, which was originally developed for detecting proteins, and by increasing the LPS oxidation time (from 5 to 20 min), we restored the ability to detect LPS fractions that otherwise would not be stained. These modifications did not affect the detection of other S- and R-form LPSs. Thus, differences in the number of fatty acids present in polyagglutinable P. aeruginosa LPS may result in a selective loss of fatty acid-deficient S-form LPS in these apparent R-form LPS preparations. This modified procedure provides a fast, simple, and sensitive way to analyze LPS in polyacrylamide gels despite the number of acyl groups present.
Profiles of serological reactivity against cytosoluble antigens of Brucella ovis in experimentally infected rams
Authors: Chin J C , Pang-Turner B
Published: January 2020

Sera from rams infected with and excreting Brucella ovis in the semen (shedders), as well as from animals which had recovered from previous experimental challenge with B. ovis, were analyzed for their serological reactivities against cytosolic antigens of the bacterium. Membrane vesicles, including outer and inner membrane components, were precluded from the analyses by subjecting French-pressed bacteria to ultracentrifugation. The resulting cytosolic supernatant was fractionated into four major antigenic fractions, fractions A, B, C, and D, by high-pressure liquid chromatography. Temporal enzyme-linked immunosorbent assays with the A antigen revealed that all shedder rams displayed a rise-and-surge response, while rams which recovered from experimental challenge showed a rise-and-fall profile. The B antigen was less discriminatory in detecting a difference between the two ram groups, while C and D antigens were serologically unreactive in the enzyme-linked immunosorbent assay. In contrast to the reactivity patterns shown by native high-pressure liquid chromatography-fractionated cytosolic supernatant antigens, immunoblotting of C and D polypeptides generated by boiling in the presence of sodium dodecyl sulfate and mercaptoethanol was particularly useful in distinguishing between sera collected at the mid-surge phase of infected rams from sera obtained at the mid-fall stage of recovered animals. It is likely that native or denatured antigens of different cytosolic fractions may provide useful serological reagents for differentiating between infected rams and those which have recovered from exposure to B. ovis.
Development and evaluation of a human T-cell leukemia virus type I serologic confirmatory assay incorporating a recombinant envelope polypeptide
Authors: Lillehoj E P , Alexander S S , Dubrule C J , Wiktor S , Adams R , Tai C C , Manns A , Blattner W A
Published: January 2020

A recombinant protein derived from the gp21 region of the human T-cell leukemia virus type I (HTLV-I) env gene was synthesized in Escherichia coli and purified by reversed-phase high-performance liquid chromatography. The purified protein was free of contaminating bacterial proteins and retained reactivity with human HTLV-I- and HTLV-II-positive sera and a gp21 monoclonal antibody. An immunoblot procedure using the recombinant polypeptide in conjunction with native viral proteins was more sensitive than the conventional immunoblot and radioimmunoprecipitation confirmatory assays for detection of antibodies to HTLV-I and HTLV-II env-encoded gene products. The recombinant protein was equally reactive with sera from polymerase chain reaction-confirmed HTLV-I or HTLV-II infections. Furthermore, on the basis of the differential reactivities of gp21-positive sera with the HTLV-I p19 and p24 gag-encoded proteins, an algorithm was proposed to distinguish exposure to HTLV-I from exposure to HTLV-II. These results establish the utility of a modified immunoblot assay incorporating a recombinant envelope polypeptide as an alternative to existing HTLV-I-confirmatory assays.
Characterization of a Mycobacterium tuberculosis insertion sequence, IS6110, and its application in diagnosis
Authors: Thierry D , Brisson-Noël A , Vincent-Lévy-Frébault V , Nguyen S , Guesdon J L , Gicquel B
Published: January 2020

An insertion sequence-like element, IS6110, was isolated from a Mycobacterium tuberculosis cosmid library as a repetitive sequence. IS6110 shows similarities with elements of the IS3 family. This insertion sequence was found to be specific to mycobacteria belonging to the M. tuberculosis complex. For detection and identification of M. tuberculosis bacilli in uncultured specimens, oligonucleotides derived from the IS6110 sequence were used as primers and probes in polymerase chain reaction studies. The results obtained were consistent with results of classical identification procedures, bacteriological data, and clinical criteria.
V factor-dependent members of the family Pasteurellaceae in the porcine upper respiratory tract
Authors: Møller K , Kilian M
Published: January 2020

A study was performed to obtain a better understanding of the diversity and ecology of members of the family Pasteurellaceae in the porcine respiratory tract. A collection of 132 V factor-dependent strains of Pasteurellaceae selected from porcine field isolates mainly from the respiratory tract were subjected to detailed characterization. In addition to the three hitherto recognized species Actinobacillus pleuropneumoniae, Haemophilus parasuis, and Haemophilus taxon "minor group," three distinct taxa were observed. Some of these taxa, which are provisionally designated taxa D, E, and F, would by traditional criteria be mistaken for H. parasuis but differed by several biochemical characteristics. To study the ecology of the V factor-dependent species, swabs from the nasal and oral cavities of 29 pigs were cultivated on selective and nonselective media. By studying approximately 30 isolates from each sample, the distribution and relative proportion of the individual taxa were determined. A. pleuropneumoniae was detected in samples from the tonsil areas of only two acutely ill animals. H. parasuis was isolated from the nasal cavities of four out of nine healthy pigs but from the oral cavities of only two animals. In contrast, taxon "minor group" and taxa D, E, and F were present in the oral cavities of the majority of pigs but were not detected in samples from their nasal cavities. The results indicate that all the observed V factor-dependent species of Pasteurellaceae except A. pleuropneumoniae, are members of the resident microflora of various mucosal surfaces of the porcine upper respiratory tract.
Laboratory investigation of Acanthamoeba keratitis
Authors: Kilvington S , Larkin D F , White D G , Beeching J R
Published: January 2020

Following the diagnosis of Acanthamoeba keratitis in a contact lens wearer, the antimicrobial susceptibility of the clinical isolate and the environmental source of the infection were investigated. Contrary to previous reports, in vitro antimicrobial testing showed that the infecting strain was inherently resistant to propamidine isethionate. Restriction endonuclease digestion analysis of Acanthamoeba whole-cell DNA of strains isolated from the patient's cornea, contact lens storage container, saline rinsing solution, and kitchen cold-water tap showed that the isolates were identical. This implicates, for the first time, domestic tap water as the source of Acanthamoeba sp. in this infection. It is therefore recommended that the use of homemade saline solutions and the rinsing of contact lenses in tap water be strongly discouraged.
Rapid detection of human papillomavirus in cervical scrapes by combined general primer-mediated and type-specific polymerase chain reaction
Authors: van den Brule A J , Meijer C J , Bakels V , Kenemans P , Walboomers J M
Published: January 2020

A two-step polymerase chain reaction (PCR) procedure was used as a new screening strategy for the detection of human papillomavirus (HPV) genotypes in cervical scrapes omitting prior DNA extraction. Sample preparation consisted of a freeze-thaw step followed by boiling the cells before the PCR mixture was added. This pretreatment was as efficient and reproducible for HPV DNA amplification as DNA purification. By using crude cell suspensions, a prescreening of the samples with the general primer-mediated PCR method (GP-PCR) was performed to detect a broad spectrum of sequenced and still unsequenced HPV types at the subpicogram level. HPV-containing scrapes by GP-PCR were subjected to HPV 6, 11, 16, 18, 31, and 33 type-specific PCR (TS-PCR) to identify the sequenced HPV types. This direct GP/TS-PCR method was tested on a large group of cervical scrapes (n = 459) from women visiting a gynecologic outpatient clinic. The results were compared with HPV data obtained by a method using modified filter in situ hybridization and TS-PCR in which the PCR was mainly used to confirm HPV positivity. A substantially higher HPV prevalence rate was found by direct GP/TS-PCR strategy. The results indicate that GP/TS-PCR is a rapid, sensitive, and reliable detection method for HPV in cervical scrapes. The easy performance on crude cell suspensions makes this strategy applicable for large HPV-screening programs.
Monoclonal antibody-based capture enzyme immunoassays for specific serum immunoglobulin G (IgG), IgA, and IgM antibodies to respiratory syncytial virus
Authors: Erdman D D , Anderson L J
Published: January 2020

Monoclonal antibodies (MAbs) to the fusion protein (F), attachment protein (G), and nucleoprotein (N) of respiratory syncytial (RS) virus were evaluated for use as detector antibodies in immunoglobulin G (IgG), IgA, and IgM capture enzyme immunoassays. MAb assays were tested against assays using polyclonal antibodies (PAbs) with serum specimens from patients with and without evidence of recent RS virus infection. Assays developed with N MAbs were comparable to or better than PAb assays for detecting specific IgG and IgM antibodies but were somewhat less sensitive for IgA. F MAb assays were less sensitive for IgG and IgM antibodies but identified specific IgA in some specimens negative by N MAb assay. G MAb assays were insensitive for IgG and IgM antibodies but did detect about 50% of the IgA antibodies identified by the PAb assay. The basis for the low sensitivity of the G MAb assays is unclear, since many of these specimens were positive for IgG antibodies to G by Western immunoblot. The sensitivity of MAb assays varied with patient age: N MAb assays detected specific antibody responses to RS virus in all immunoglobulin classes in both adults and infants less than 1 year of age, F MAb assays detected specific IgG responses in adults and IgA responses in both adults and infants, and G MAb assays only detected IgA responses in adults. A mixture of N and F MAbs was complementary overall, identifying 54 of 55 (IgG), 51 of 52 (IgA), and 16 of 17 (IgM) serum specimens positive by PAb assay. These MAb assays were also specific with specimens tested from persons without a history of recent RS virus infection. The availability of these MAb-based assays offers other laboratories the opportunity to have long-term, standardized reagents and tests for serological diagnosis of RS virus infection.
Leptomyxid ameba, a new agent of amebic meningoencephalitis in humans and animals
Authors: Visvesvara G S , Martinez A J , Schuster F L , Leitch G J , Wallace S V , Sawyer T K , Anderson M
Published: January 2020

Amebae belonging to the order Leptomyxida are regarded as innocuous soil organisms incapable of infecting mammals. We report here the isolation of a leptomyxid ameba from the brain of a pregnant baboon (Papio sphinx) that died of meningoencephalitis at the San Diego Zoo Wild Animal Park. By using rabbit anti-leptomyxid serum in the immunofluorescence assay, we have identified the leptomyxid ameba in the brain sections of a number of human encephalitic cases from around the world as well as a few cases of meningoencephalitis in animals in the United States, which suggests that the leptomyxid amebae are potential etiologic agents of fatal meningoencephalitis in humans and animals.
Association of coagulase-negative staphylococcal slime production and adherence with the development and outcome of adult septicemias
Authors: Kotilainen P
Published: January 2020

The relationship of coagulase-negative staphylococcal slime production and adherence with the development and outcome of bloodstream infections in two Finnish hospitals was evaluated. Analysis of 64 strains from 62 adult septicemias disclosed 34 (53%) adherent slime producers. In comparison, only 142 (29%) of 489 single blood culture isolates were adherent slime producers. Although tube adherence test-positive strains were significantly (P less than 0.001) more common among the septicemia strains than among clinically insignificant isolates, almost half of the septicemia cases were caused by tube test-negative strains. Thus, regarding any single patient isolate, a cautious posture to the clinical impact of positivity in the tube adherence test seems warranted. Moreover, adherence and slime production, as such, apparently played no role in the clinical outcome of these infections. The epidemiologic findings revealed that slime-producing coagulase-negative staphylococci were common in the hospital environment and suggested that epidemic spread of such strains was influenced by antimicrobial therapy. Collectively, these results indicate that, at least in these two hospitals, positivity in the tube was of minor importance in guiding clinical decisions in treating adult septicemias.
Prevalent enteric adenovirus variant not detected by commercial monoclonal antibody enzyme immunoassay
Authors: Scott-Taylor T , Ahluwalia G , Klisko B , Hammond G W
Published: January 2020

A commercial monoclonal antibody enzyme immunoassay for the detection of enteric adenovirus type 40 and 41 (Ad40 and Ad41) in stool specimens was evaluated. Twenty-one stool specimens from children with gastroenteritis, with adenovirus particles visible by electron microscopy, and reference strains Ad40 Dugan and Ad41 Tak were tested by Ad40- and Ad41-specific and adenovirus group-reactive immunoassays. All stool specimens tested positive in the group-reactive immunoassay. However, only six specimens, containing isolates of Ad40 strain Hovi-X, an Ad40 genomic variant, and Ad41 strain Tak, reacted with the specific immunoassay, besides the reference strains. Fifteen stool specimens determined by restriction analysis to contain a genomic variant of Ad41 were negative by specific immunoassay. The positions of restriction site differences from the prototype strain Ad41 Tak were analyzed, and four mutations were mapped within the hexon gene; two others may occur in the fiber gene. The Ad41 genomic variant not detected by the enteric test is presently the most frequent cause of local adenoviral gastroenteritis. Highly specific monoclonal antibodies can fail to detect genomic variants of enteric adenoviruses, probably because of alteration of external neutralizable epitopes under immunological pressure to vary.
Evaluation of a novel solid-phase immunoassay, Clearview Chlamydia, for the rapid detection of Chlamydia trachomatis
Authors: Arumainayagam J T , Matthews R S , Uthayakumar S , Clay J C
Published: January 2020

Clearview Chlamydia (Unipath Limited, Bedford, United Kingdom) is a rapid immunoassay for the direct detection of Chlamydia trachomatis antigen. This assay was evaluated against the tissue culture method by using 376 paired endocervical specimens. The Clearview assay had a sensitivity of 93.5% and a specificity of 99% when it was compared with the tissue culture method. This assay does not require specialized equipment or trained personnel and yields results within 30 min from the time that a specimen is collected.
Labile type-specific antigen of Moraxella catarrhalis
Authors: Norkus S J , Vennes J W
Published: January 2020

By using hot acid extract and double-diffusion studies, an antigen isolated from clinical strains of Moraxella catarrhalis, designated the C antigen, was studied. The antigen is labile, relatively trypsin insensitive, and either polysaccharide or glycoconjugate in nature. At least two serologically distinct C antigens have been identified.
Anaerobic endocarditis caused by Staphylococcus saccharolyticus
Authors: Westblom T U , Gorse G J , Milligan T W , Schindzielorz A H
Published: January 2020

The first case of infective endocarditis caused by the anaerobe Staphylococcus saccharolyticus is reported. The infection occurred in a previously healthy 61-year-old male with no known predisposing valvular heart disease. The patient was successfully treated with a combination of 2 g of nafcillin every 4 h and 90 mg of gentamicin every 8 h for 6 weeks.
Patterns of morphologic variation among isolates of Trichosporon beigelii
Authors: Lee J W , Melcher G A , Rinaldi M G , Pizzo P A , Walsh T J
Published: January 2020

We observed considerable variability in colony and microscopic morphology among isolates of Trichosporon beigelii. Deeply invasive clinical isolates showed four distinct morphotypes and spontaneous conversions among certain morphotypes and grew well at 37 degrees C. In contrast, superficial clinical and environmental isolates did not demonstrate such morphotypes or conversions, and most grew poorly at 37 degrees C. Thus, the morphologic and physiologic features of invasive clinical isolates of T. beigelii follow certain patterns distinct from those of superficial clinical and environmental isolates.
Hepatitis B virus replication within the human spleen
Authors: Di Bisceglie A M , Hoofnagle J H
Published: January 2020

Southern blot analysis of splenic DNA from three patients with chronic hepatitis B revealed the presence of replicative intermediate forms of viral DNA. Hepatitis B viral RNA was detected in splenic RNA from two cases. Thus, hepatitis B virus appears to replicate in the human spleen, although at a lower level than in the liver.
Conversion of Pseudomonas aeruginosa to the phenotype characteristic of strains from patients with cystic fibrosis
Authors: Speert D P , Farmer S W , Campbell M E , Musser J M , Selander R K , Kuo S
Published: January 2020

Isolates of Pseudomonas aeruginosa from cystic fibrosis patients are unusual; they are often susceptible to the bactericidal effect of human serum, have a rough lipopolysaccharide, and produce an exopolysaccharide that is responsible for the characteristic mucoid phenotype. In contrast, strains from the environment and from patients with other diseases usually have smooth lipopolysaccharide, do not produce very much mucoid exopolysaccharide, and are phenotypically nonmucoid. The predominance of mucoid strains of P. aeruginosa in infections of patients with cystic fibrosis has not been explained. In the lower airways, where P. aeruginosa persists in cystic fibrosis, nutrients for bacterial growth may be limited. We investigated whether growth of P. aeruginosa under conditions of suboptimal nutrition causes conversion to the characteristic cystic fibrosis phenotype. Ninety-two strains of P. aeruginosa were maintained for up to 90 days in a minimal medium with acetamide as the sole carbon source. In 56 (52%) of 107 cultures, isolates with rough lipopolysaccharide emerged, and in 20 (19%) of 104 nonmucoid cultures, mucoid isolates were recovered. Strains with rough lipopolysaccharide also were sensitive to the bactericidal effect of normal human serum. Under conditions of suboptimal nutrition in vitro, isolates of P. aeruginosa emerged that produced rough lipopolysaccharide and were mucoid, typical of many isolates from cystic fibrosis patients. This peculiar phenotype may arise as a consequence of nutritional limitation within the cystic fibrosis respiratory tract rather than from features unique to these strains of bacteria.
Effect of primary Epstein-Barr virus infection on human herpesvirus 6, cytomegalovirus, and measles virus immunoglobulin G titers
Authors: Linde A , Fridell E , Dahl H , Andersson J , Biberfeld P , Wahren B
Published: January 2020

Immunoglobulin G antibody titers to human herpesvirus 6 (HHV-6), measles virus, and cytomegalovirus (CMV) were examined in serum samples from 31 patients with Epstein-Barr virus (EBV)-induced infectious mononucleosis (IM). Sera were drawn sequentially from the same patients less than or equal to 7 days until 3 years after onset of IM. In seropositive patients, there was a significant decrease with time after IM of the immunoglobulin G titers to the three viruses in the majority of patients; HHV-6 IgG titers decreased in 80%, measles virus IgG titers decreased in 75%, and CMV IgG titers decreased in 67%. Four patients contracted CMV infection during the observation period after IM. In these, HHV-6 IgG titers increased, while EBV and measles virus IgG titers remained essentially stationary. Polyclonal B-cell stimulation during IM is suggested to augment antiviral titers in general, but the increases of HHV-6 IgG titers during EBV and CMV infections may also be due to selective stimulation of memory B cells by related antigens or to reactivation of HHV-6 during infection with these herpesviruses.
Evaluation of three Chlamydia trachomatis immunoassays with an unbiased, noninvasive clinical sample
Authors: Paul I D , Caul E O
Published: January 2020

First-catch early morning urine samples from patients attending a genitourinary medicine clinic were tested by three different enzyme immunoassays. The results suggest significant differences in the sensitivities and specificities of the different assays. The direct visualization of elementary bodies in urine deposits by direct immunofluorescence was used as the "gold standard," using a monoclonal antibody with a different epitope specificity from those of antibodies used in the enzyme-linked immunosorbent assays. We report for the first time that urine specimens represent an unbiased sample, free of the inherent sampling errors associated with other genital specimens. We feel that urine is a valid specimen for use in any evaluation study of new assays directed towards the detection of Chlamydia trachomatis.
Identification of Bacillus anthracis by using monoclonal antibody to cell wall galactose-N-acetylglucosamine polysaccharide
Authors: Ezzell J W , Abshire T G , Little S F , Lidgerding B C , Brown C
Published: January 2020

Guanidine extracts of crude Bacillus anthracis cell wall were used to vaccinate BALB/c mice and to develop monoclonal antibody (MAb) to vegetative cell surface antigens. Two hybridomas selected during this study produced immunoglobulin M immunoglobulins, which appear to be directed to an epitope associated with the galactose-N-acetyl-D-glucosamine polysaccharide. Both demonstrated specificity in their binding to purified B. anthracis cell wall, o-stearoyl-polysaccharide conjugates, and intact, nonencapsulated vegetative cells. The interaction of the MAbs with purified polysaccharide was inhibited by 0.5 M galactose and lactose but not by N-acetylglucosamine, glutamate, glycine, or glycerol. Inhibition by glucose or sucrose was approximately 75% of that seen with galactose. Electron microscopy showed that both MAbs interacted with the cell wall of vegetative cells as well as with the cortex of spores. Neither MAb reacted with encapsulated vegetative cells, such as those from infected guinea pigs, nor did they react with intact spores. After conjugation to fluorescein isothiocyanate, the MAbs stained intensely all B. anthracis strains tested, whereas with two exceptions, none of the strains of 20 other Bacillus spp. was stained. The exceptions, strains of Bacillus cereus, could be differentiated from B. anthracis by being beta-hemolytic on blood agar.
Clinical application of novel sample processing technology for the identification of salmonellae by using DNA probes
Authors: Scholl D R , Kaufmann C , Jollick J D , York C K , Goodrum G R , Charache P
Published: January 2020

Two hundred and fifty clinical fecal specimens collected over a 7-month period were analyzed for the presence of salmonellae by a rapid DNA hybridization procedure. Hybridizations were performed by using a novel specimen processing protocol called wicking and a previously unreported 1,600-base-pair probe cloned from Salmonella enteritidis DNA. The probe was shown to be reactive with all 70 Salmonella serotypes tested and not reactive with 101 stock strains of other enteric bacteria. Southern analysis of 30 Salmonella isolates representing 22 serotypes suggested that the probe sequence was highly conserved, appearing as a 1,600-base-pair band in a BglII digest of isolate DNA in 29 of 30 isolates and as a 2,300-base-pair fragment in 1 of the isolates. The probe correctly identified all salmonellae (nine isolates) among 47 H2S-producing colonies tested from among 250 clinical specimens cultured on xylose-lysine-desoxycholate medium. Salmonellae grown on xylose-lysine-desoxycholate medium gave consistently higher hybridization values than did those grown on either MacConkey or Hektoen enteric agar. In addition, of eight gram-negative broth enrichments in which salmonellae were identified by conventional means, seven were probe positive. The use of this nucleic acid probe and hybridization technique provides a simple and rapid identification of Salmonella species.
Molecular typing of nosocomial isolates of Legionella pneumophila serogroup 3
Authors: Tram C , Simonet M , Nicolas M H , Offredo C , Grimont F , Lefevre M , Ageron E , Debure A , Grimont P A
Published: January 2020

In Paris, France, an outbreak of pneumonia due to Legionella pneumophila serogroup 3 was observed in Necker (four cases) and Pitié (six cases) hospitals. Neither the 10 clinical isolates nor 5 tap water isolates from Necker Hospital harbored plasmids. Clinical and environmental serogroup 3 isolates and serogroup 3 reference strain Bloomington 2 were analyzed by chromosomal probe fingerprinting. rRNA, 16S and 23S from Escherichia coli and a randomly cloned 15-kilobase-pair nucleotide sequence from L. pneumophila serogroup 3 were used as probes. All strains tested showed a single pattern after HindIII digestion of DNA and hybridization with the 32P-end-labeled rRNA probe, whereas three patterns were obtained after hybridization with the 32P-labeled 15-kilobase-pair DNA probe. One pattern was given by all clinical and tap water isolates from Necker Hospital, another one was given by all clinical isolates from Pitié Hospital, and a last one was given by reference strain Bloomington 2. Thus, molecular analysis showed that the two hospital outbreaks of legionellosis were unrelated and could link the outbreak in Necker Hospital to contaminated tap water.
Collaborative evaluation of antigen detection by a commercial latex agglutination test and enzyme immunoassay in the diagnosis of invasive candidiasis
Authors: Lemieux C , St-Germain G , Vincelette J , Kaufman L , de Repentigny L
Published: January 2020

The Cand-Tec Candida detection system and enzyme immunoassay for serum mannan were retrospectively compared in a controlled collaborative evaluation of antigen detection in 32 patients with candidiasis proven by biopsy or culture from a normally sterile site and with sera drawn within 7 days of inclusion. With a threshold titer of 1/8, which excluded false-positive results in 17 hospitalized patients without candidiasis, sensitivities for all 32 patients with candidiasis were 44% for the Cand-Tec assay and 17% for the enzyme immunoassay. Both assays provided greater sensitivity when sera were drawn within 24 h of inclusion in the study and in the category of patients with invasive candidiasis (57% by Cand-Tec and 33% by enzyme immunoassay). The Cand-Tec assay gave false-positive results (titer, greater than or equal to 1/8) in 4 of 6 patients with transient candidemia, in 1 of 20 otherwise healthy patients with rheumatoid factor, and in 1 patient with a positive cryptococcal latex agglutination test. Three serum specimens from 3 of 32 patients with candidiasis contained rheumatoid factor and gave titers of greater than or equal to 1/8 by the Cand-Tec assay. Detection of serum mannan by enzyme immunoassay was less sensitive but more specific than the Cand-Tec Candida detection system for the diagnosis of invasive candidiasis.
Biochemical identification of Aeromonas genospecies isolated from humans
Authors: Altwegg M , Steigerwalt A G , Altwegg-Bissig R , Lüthy-Hottenstein J , Brenner D J
Published: January 2020

One hundred phenotypic characteristics were determined for 138 clinical and environmental Aeromonas strains. Cluster analysis revealed three major phenons equivalent to the A. hydrophila, A. caviae, and A. sobria groups, each of which contained more than one genospecies and more than one named species. An excellent correlation was found between phenotypic identification and classification based on DNA relatedness. DNA hybridization groups within each of the phenotypic groups were also separable by using a few biochemical characteristics. Key tests were production of acid from or growth on D-sorbitol (which separated DNA hybridization group 3 from groups 1 and 2 within the A. hydrophila phenogroup), growth on citrate (which essentially separated DNA hybridization group 4 from groups 5A and 5B within the A. caviae phenogroup), and growth on DL-lactate (which separated DNA hybridization group 1 from groups 2 and 3 within the A. hydrophila phenogroup as well as group 5A from groups 4 and 5B within the A. caviae phenogroup). All except one strain in the A. sobria phenogroup belonged to DNA hybridization group 8. DNA hybridization groups were not equally distributed among clinical and environmental isolates, suggesting that strains of certain DNA hybridization groups might be less virulent than others.