bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigenuria in infants with acute and congenital Chagas' disease
Authors: Freilij H L , Corral R S , Katzin A M , Grinstein S
Published: January 2020

Detection and partial characterization of Trypanosoma cruzi soluble antigens (SAg) in urine, as well as demonstration of parasite circulating antigens (CAg) in serum from pediatric patients with acute (10 patients) and congenital (10 patients) Chagas' disease, are reported. Classical techniques for parasite detection and antibody serology were also conducted in both groups. Samples collected before the onset of parasiticidal drug treatment were tested by an enzyme-linked immunosorbent assay for SAg and CAg demonstration. The control population consisted of 6 children with acute toxoplasmosis, 6 with cutaneous leishmaniasis, and 20 healthy individuals. Patients with acute cases were 100% positive for both SAg and CAg, whereas patients with congenital disease were 80% CAg positive and 100% SAg positive. Controls yielded negative results in all cases. Partial characterization of SAg from two patients with acute disease was performed by iodination, affinity chromatography, immunoprecipitation, and two-dimensional gel electrophoresis. Two different antigenic glycoproteins (80 kilodaltons, pI 6 to 6.5 and 55 kilodaltons, pI 6.5 to 7) were identified by these methods. Traditional serology and classical parasitologic tests failed, each in a different way, to provide an accurate diagnosis in the total of our patients. The enzyme-linked immunosorbent assay for SAg detection proved to be the most effective procedure for achieving early and precise proof of infection in acute and congenital cases of Chagas' disease.
Performance of four slide agglutination methods for identification of Staphylococcus aureus when testing methicillin-resistant staphylococci
Authors: Lairscey R , Buck G E
Published: January 2020

Four commercially available slide agglutination systems for identifying Staphylococcus aureus were compared with the conventional slide (clumping factor) and tube coagulase tests. The systems evaluated included Bacto Staph Latex (Difco Laboratories, Detroit, Mich.), Staphyloslide (BBL Microbiology Systems, Cockeysville, Md.), Mini ID Accu-Staph (Carr-Scarborough Microbiologicals, Inc., Decatur, Ga.), and Staphaurex (Wellcome Diagnostics, Research Triangle Park, N.C.). A total of 338 clinical isolates, including methicillin-resistant S. aureus (n = 149), methicillin-susceptible S. aureus (n = 78), methicillin-resistant, coagulase-negative staphylococci (n = 45), and methicillin-susceptible, coagulase-negative staphylococci (n = 66), were tested by each method. The slide test for clumping factor, the 4-h tube coagulase test, Bacto Staph Latex, Staphyloslide, Mini ID Accu-Staph, and Staphaurex detected 212 (93.4%), 218 (96%), 223 (98.2%), 223 (98.2%), 221 (97.4%), and 224 (98.7%) of the S. aureus (44% methicillin-resistant) isolates, respectively. There were no false-positive results with any of the methods when the 111 strains of coagulase-negative staphylococci were tested. The results of this evaluation suggest that the four slide identification methods tested can provide rapid and accurate identification of methicillin-resistant S. aureus strains.
Clinical evaluation of the Vitek Neisseria-Haemophilus Identification card
Authors: Janda W M , Malloy P J , Schreckenberger P C
Published: January 2020

A clinical evaluation of the Vitek Neisseria-Haemophilus Identification (NHI) card (Vitek Systems, Inc., Hazelwood, Mo.) was performed with 480 clinical isolates and stock strains of Neisseria spp., Haemophilus spp., and other fastidious microorganisms included in the data base of the system. Identifications obtained with the NHI card were compared with those determined by conventional methods. The card identified 83.2% of 244 Neisseria spp. and Branhamella catarrhalis, 54.9% of 164 Haemophilus spp., and 84.7% of 72 fastidious gram-negative species with no further testing required. Some isolates produced good confidence-marginal separation identifications, in which the correct identification was listed with one or two other possible identifications and extra tests were required and suggested. When isolates producing good confidence-marginal separation identifications were included, correct identifications of these organism groups increased to 97.1, 92.7, and 94.4%, respectively. Among the commonly isolated microorganisms, the NHI card identified 99.1% of 110 N. gonorrhoeae, 98.5% of 68 N. meningitidis, 93.9% of 98 H. influenzae, and 95.6% of 46 H. parainfluenzae strains. All of these organisms produced excellent to very good confidence level identifications except for H. influenzae biotypes II, III, and VII, for which hemolytic reactions were required for differentiation from H. haemolyticus. The NHI card reliably identified other fastidious gram-negative species, including H. aphrophilus, Eikenella corrodens, Gardnerella vaginalis, and Kingella denitrificans.
Investigation of association of mycobacteria with inflammatory bowel disease by nucleic acid hybridization
Authors: Yoshimura H H , Graham D Y , Estes M K , Merkal R S
Published: January 2020

We used DNA-DNA hybridization to characterize a mycobacterial isolate, strain Linda, that was obtained from a patient with Crohn's disease and that has been reported to cause ileitis in experimental animals. We also investigated the association of this mycobacterium with Crohn's disease. Our results identified Mycobacterium strain Linda as a strain of Mycobacterium paratuberculosis, the etiologic agent of Johne's disease, a disease of ruminants that has some superficial resemblance to Crohn's disease. Sequences that hybridized with strain Linda DNA probes were detected in DNA extracted from human intestinal tissues from patients with Crohn's disease, ulcerative colitis, and noninflammatory bowel disease. These hybridizing DNA sequences were more prevalent in the muscle layers than in the intestinal mucosa, making it unlikely that they represented DNA from bacterial contaminants in the intestinal lumen. Measurement of the melting temperatures of the DNA-DNA hybrids formed between strain Linda probes and tissue DNAs indicated that the related sequences detected were of mycobacterial origin but were not identical to each other or to strain Linda DNA. These results do not support the proposed specific relationship between Mycobacterium strain Linda and Crohn's disease. The possible etiologic role of mycobacteria in Crohn's disease is discussed.
Variability of cell surface hydrophobicity among Pasteurella multocida somatic serotype and Actinobacillus lignieresii strains
Authors: Darnell K R , Hart M E , Champlin F R
Published: January 2020

Pasteurella multocida possesses a characteristically gram-negative ultrastructure, yet its inability to grow in the presence of hydrophobic compounds and the general penicillin susceptibility of genera making up the family Pasteurellaceae suggest a cell envelope having atypical permeability properties. The cell surface hydrophobicity properties of strains representing 15 of the 16 somatic serotypes of P. multocida and three strains of Actinobacillus lignieresii were assessed with hydrocarbon adherence and hydrophobic interaction chromatographic assays. These methods revealed surface hydrophobicity to vary dramatically among strains in both species. No direct correlation was observed with species, growth rate, or susceptibility to the antibiotics oxytetracycline (polar), polymyxin B (amphiphilic), or novobiocin (nonpolar) as measured with MIC determinations. All strains were susceptible to the antibiotics, although A. lignieresii was significantly less susceptible than P. multocida to novobiocin. These data suggest that cell surface hydrophobicity in P. multocida may be influenced by the type of lipopolysaccharide present but is not directly related to permeability of the antibiotics examined. The wide diversity of hydrophobic properties exhibited by strains of both P. multocida and A. lignieresii precludes the use of this parameter as a taxonomic acid.
Comparative sensitivity of 125I-protein A and enzyme-conjugated antibodies for detection of immunoblotted proteins
Authors: Bernstein J M , Stokes C E , Fernie B
Published: January 2020

Immunoblotting is a powerful technique for the detection of small amounts of immunologically interesting proteins in unpurified preparations. Iodinated protein A (PA) has been widely used as a second antibody for detection of proteins; however, it does not bind equally well to immunoglobulins from different species nor does it bind to all subclasses of immunoglobulin G (IgG). We compared the sensitivity of [125I]PA with those of both horseradish peroxidase-conjugated second antibodies (HRP) and glucose oxidase-anti-glucose oxidase (GAG) soluble complexes for visualizing bovine serum albumin, human IgG, or human C3 which was either dot blotted or electroblotted to nitrocellulose. [125I]PA was uniformly 10- to 100-fold less sensitive than either HRP or GAG. GAG was more sensitive than HRP except for C3 (electroblotting) and bovine serum albumin and IgG (dot blotting), in which they were equivalent. In general, dot blotting was 10- to 1,000-fold more sensitive than electroblotting. Although relative sensitivities varied depending on the proteins analyzed and the antisera used, GAG appeared to be superior to [125I]PA and HRP for detection of immunoblotted proteins.
Production, characterization, and species specificity of five monoclonal antibodies to Mycobacterium tuberculosis
Authors: Kadival G V , Chaparas S D
Published: January 2020

The production and characterization of five monoclonal antibodies to Mycobacterium tuberculosis are described. Specificity of the monoclonal antibodies was tested against other mycobacterial species by an enzyme-linked immunosorbent assay and immunoblots. HGT 3a, an immunoglobulin M (IgM) antibody, recognizes a molecule of 38,000 molecular weight present only in the tuberculosis complex of M. tuberculosis and Mycobacterium bovis BCG. HGT 6, an IgG1 antibody, recognized two molecules with molecular weights of 43,000 and 45,000 and showed limited cross-reactivity. Three other antibodies, HGT 1, HGT 2, and HGT 4, all belonging to the IgG1 type, recognized multiple bands and showed broad reactivity among all mycobacterial antigens tested, Escherichia coli and Nocardia asteroides.
Detection of human antibodies against Plasmodium falciparum sporozoites using synthetic peptides
Authors: Del Giudice G , Verdini A S , Pinori M , Pessi A , Verhave J P , Tougne C , Ivanoff B , Lambert P H , Engers H D
Published: January 2020

A large peptide consisting of about 40 (Asn-Ala-Asn-Pro) repeats of Plasmodium falciparum circumsporozoite protein, (NANP)40, was synthesized. It was recognized specifically by monoclonal antibodies produced against P. falciparum sporozoites. Moreover, this peptide strongly inhibited the binding of such monoclonal antibodies to antigens present in a sporozoite extract. The (NANP)40 peptide was employed without any carrier to develop an enzyme-linked immunosorbent assay to detect sporozoite-specific serum antibodies arising after natural malaria infections. Antibodies were detected in a high percentage (43.1%) of European patients suffering from acute P. falciparum malaria and in Africans living in an area of Gabon endemic for malaria. In the latter group, the frequency of antisporozoite antibodies increased with age, reaching 65.9% in individuals more than 40 years old. There was a significant correlation between the results obtained with an immunofluorescence assay with glutaraldehyde-fixed sporozoites and those obtained by enzyme-linked immunosorbent assay with (NANP)40. Therefore, such synthetic peptides representing the repetitive epitope of P. falciparum circumsporozoite protein can be used for the detection of antisporozoite antibodies and for the epidemiological studies required to obtain base-line data concerning the immune status of individuals before their participation in a sporozoite vaccine trial.
Humoral immune response to polypeptides of Ureaplasma urealyticum in women with postpartum fever
Authors: Lee G Y , Kenny G E
Published: January 2020

We investigated the antibody response in women with postpartum fever from whom Ureaplasma urealyticum had been isolated from the bloodstream. Acute- and convalescent-phase sera were tested for immunoglobulin G to the polypeptides of five serovars (1, 2, 3, 4, and 8), representing the two genomic clusters of U. urealyticum, by the immunoblotting (Western) method. Convalescent-phase sera from the five patients reacted more intensely and with more (up to 27) polypeptides from each of the five serovars, whereas acute-phase sera reacted weakly and with few polypeptides. Although antibody responses in these women with systemic infection could be detected by the use of any of the five different serovars as antigens, the patterns that were produced differed clearly between the two genetic clusters (serovars 1 and 3 versus serovars 2, 4, and 8). Apparently, a single serovar could be used to detect ureaplasmal antibodies in humans regardless of the serovar of the infecting strain.
Failure of multiple passages to increase chlamydial recovery
Authors: Schachter J , Martin D H
Published: January 2020

Because of a previous publication indicating that increasing the number of blind passages to five resulted in a marked increase in Chlamydia trachomatis isolation rates, we evaluated a multiple-blind-passage system in vials. Cervical specimens from 119 pregnant women and 82 specimens from other sources were tested for chlamydial infection. The recovery rate for C. trachomatis was not markedly increased after the first blind passage. Thus, we conclude that multiple blind passages are not necessary to maximize the C. trachomatis isolation rate in a vial system.
Preparation by ultrafiltration and control by high-performance liquid chromatography of the native hapten of Brucella abortus for use in radial immunodiffusion diagnostic test
Authors: Zygmunt M S , Dubray G
Published: January 2020

An effective method was developed for the preparation of Brucella abortus native hapten by use of an ultrafiltration system. The membranes PM30 and YM5 were used to separate the lipopolysaccharide and the native hapten. The yield of the native hapten was higher than that obtained by a more complex procedure reported previously. This method is economical for the large-scale preparation of B. abortus native hapten. The effects of ultrafiltration were evaluated by a quick and sensitive high-performance liquid chromatographic technique.
Detection of chlamydial inclusions in cell culture or biopsy tissue by alkaline phosphatase-anti-alkaline phosphatase staining
Authors: Mahony J B , Sellors J , Chernesky M A
Published: January 2020

An immunological technique for detecting Chlamydia trachomatis and Chlamydia psittaci inclusions in infected McCoy cell cultures was developed by using a genus-specific monoclonal antibody to Chlamydia spp., rabbit anti-mouse immunoglobulin G bridging antibody, alkaline phosphatase-anti-alkaline phosphatase (APAAP) monoclonal antibody conjugate, and naphthol AS-phosphate/fast red substrate. Chlamydial inclusions stained red and were easily detected against a background of blue hematoxylin-stained nuclei. After 18 h, inclusions of C. trachomatis serovar L2 LGV434/Bu and C. psittaci strain 6BC were stained by APAAP but not by iodine or Giemsa. At 48 h inclusion counts were significantly higher in the APAAP cultures. Both the APAAP procedure and conventional staining detected 35 of 239 (15%) cultures 48 h after inoculation with urethral or endocervical specimens. However, at 24 h after inoculation 22 of 35 (63%) were positive by APAAP staining while negative by iodine. This immunostain also allowed identification of chlamydial inclusions in endometrial biopsies from patients with tubal factor infertility or pelvic inflammatory disease.
Antigenic characterization of rotaviruses isolated in Kenya from 1982 to 1983
Authors: Urasawa T , Urasawa S , Chiba Y , Taniguchi K , Kobayashi N , Mutanda L N , Tukei P M
Published: January 2020

The electropherotypes of human rotavirus RNAs from 100 diarrheic stool specimens collected in two major districts of Kenya from 1982 to 1983 were previously reported (Y. Chiba, C. Miyazaki, Y. Makino, L. N. Mutanda, A. Kibue, E. O. Lichenga, and P. M. Tukei, J. Clin. Microbiol. 19:579-582, 1984). Of these specimens, 25 that contained rotaviruses with different RNA electropherotypes were subjected to a virus isolation experiment with MA-104 cells, and 16 rotavirus strains were isolated. The use of an enzyme-linked immunosorbent assay with subgroup-specific monoclonal antibodies enabled us to successfully subgroup 15 isolates: 4 in subgroup I and 11 in subgroup II. By fluorescent-focus-neutralization test with serotype-specific rabbit antisera, 13 isolates could be serotyped: 7 as serotype 1, 4 as serotype 2, and 2 as serotype 3. Of the remaining three isolates, F153, F247, and G402, the former was doubly neutralizable with serotype 1 and serotype 4 antisera and the latter two were neutralizable with serotype 3 and serotype 4 antisera. Detailed analysis with the antisera against F153 and F247 and four serotype-specific, VP7-directed monoclonal antibodies suggested that F153 is a serotypic mosaic strain with serotype 4-specific VP3 and serotype 1-specific VP7 outer capsid proteins and F247 and G402 are possibly antigenic mosaic strains with serotype 3 and serotype 4 antigens. On the basis of the correspondence of the rotavirus isolate serotypes determined in this study to the electropherotypes reported previously, it was inferred that serotype 1 strains were most prevalent in two districts of Kenya from 1982 to 1983, followed by any type of serotypic mosaic strains.
Toxoplasma gondii antigens recognized by sequential samples of serum obtained from congenitally infected infants
Authors: Potasman I , Araujo F G , Thulliez P , Desmonts G , Remington J S
Published: January 2020

The Sabin-Feldman dye test, the immunoglobulin M (IgM) immunosorbent agglutination assay, and the immunoblot technique were used to study the evolution of the antibody response to Toxoplasma gondii and to examine antigens of the organism recognized by antibodies in the sera of 12 congenitally infected infants and 7 mothers. In the sera of eight infants, a significant rise was noted in the dye test titers, while the serum of only one infant demonstrated a late increase in the IgM immunosorbent agglutination assay titer. In each infant and mother, antigens with approximate masses of 35,000 and 115,000 daltons were strongly recognized by IgG antibodies. An antigen(s) with an approximate mass of 4,000 daltons was recognized by IgM antibodies in the sera of each of the mothers but was recognized by antibodies in the sera of only two of the infants.
Campylobacter etiology in human gastroenteritis demonstrated by antibodies to acid extract antigen
Authors: Rautelin H I , Kosunen T U
Published: January 2020

Campylobacter antibodies of the immunoglobulin G (IgG), IgM, and IgA classes were determined by enzyme immunoassay with acid glycine extract antigen in patients and controls involved in two Campylobacter outbreaks and in 266 unselected patients with acute enteritis. The assay showed a specificity of 99% for each immunoglobulin class in sera from 200 healthy blood donors. Elevated Campylobacter antibody titers were shown in 97% of stool culture-positive patients involved in the outbreaks. Rapid changes of IgA and IgM Campylobacter antibodies were typical of the early phase of serologic response in the outbreaks and thus offered the best diagnostic value in the serologic diagnosis of acute campylobacteriosis. In unselected patients with acute enteritis, the assay revealed elevated Campylobacter antibody titers in 37 patients, of whom only 12 had had positive Campylobacter stool cultures. In the sera of patients with other bacterial findings in addition to high titers of Campylobacter antibodies, no cross-reacting antibodies were found, but there was evidence of several mixed infections.
Detection of enterotoxigenic Escherichia coli by colony hybridization with biotinylated enterotoxin probes
Authors: Kirii Y , Danbara H , Komase K , Arita H , Yoshikawa M
Published: January 2020

By using biotinylated enterotoxin DNA probes, a method to detect enterotoxigenic Escherichia coli by colony hybridization was developed. The treatment of colonies on nitrocellulose membrane filters with proteinase K and Triton X-100 was essential for obtaining the specific hybridization. A total of 200 E. coli strains isolated from travelers with diarrhea were tested for colony hybridization by using a probe encoding heat-labile toxin (LT) type h. All strains (86 of 86) that produced LT, but none of the non-LT producers, hybridized with 32P-labeled and biotinylated LT type h probes. A total of 36 strains chosen randomly from the 200 isolates were tested for colony hybridization by using heat-stable enterotoxin (ST) probes. All but two strains that hybridized with the 32P-labeled ST type Ia probe also hybridized with the biotinylated ST type Ia probe. All strains that hybridized with the 32P-labeled ST type Ib probe also hybridized with the biotinylated ST type Ib probe. Thus, almost all E. coli strains tested were judged to be the same by colony hybridization with biotinylated or 32P-labeled enterotoxin probes. These results demonstrate that the biotinylated enterotoxin probes are useful in the diagnosis of enterotoxigenic E. coli strains by colony hybridization.
Dot immunobinding assay for detection of human immunodeficiency virus-associated antigens
Authors: Blumberg R S , Hartshorn K L , Ardman B , Kaplan J C , Paradis T , Vogt M , Hirsch M S , Schooley R T
Published: January 2020

The detection of human immunodeficiency virus (HIV)-associated antigens was simplified by the application of dot immunobinding on a nitrocellulose matrix. Antigens were detected by applying the polyethylene glycol-precipitated supernatants of experimentally infected cultures directly onto nitrocellulose strips and sequentially incubating the strips with an anti-HIV antiserum and an alkaline phosphatase-conjugated, species-specific antiserum. The immune reaction was developed by adding the precipitable substrate indoyl phosphate. The dot immunobinding assay was nearly as sensitive as the reverse transcriptase assay in detecting HIV antigens in experimentally infected peripheral blood mononuclear cells, as well as in a T-cell line. The technique was also useful in the in vitro evaluation of antiviral agents. The dot immunobinding assay is a simple and sensitive technique that is useful in the detection of HIV antigens in studies of viral pathogenesis.
Densitometric analysis of Western blot (immunoblot) assays for human immunodeficiency virus antibodies and correlation with clinical status
Authors: Schmidt G , Amiraian K , Frey H , Stevens R W , Berns D S
Published: January 2020

Western blot assays for antibodies directed against components of human immunodeficiency virus (HIV) associated with acquired immunodeficiency syndrome (AIDS) were examined with a densitometer and integrator. Antibody responses to seven HIV proteins were determined from the areas under the peaks of bands on blots from 430 seropositive individuals. Antibody responses corresponded qualitatively and quantitatively with clinical status. The Western blot assays examined were done on single specimens from individuals in one of four clinical states: asymptomatic with no risk factor identified, asymptomatic with risk factor(s) identified, AIDS-related complex, and AIDS. The ratios of gp41 antibody to p24 antibody and of gp41 antibody to total HIV antibodies increased, and the number of total HIV antibodies decreased progressively in these populations. Parameters were assigned to characterize the typical response found in AIDS: gp41 antibody/p24 antibody ratio, greater than or equal to 2.0; gp41 antibody/total HIV antibodies ratio, greater than or equal to 0.30; and number of total HIV antibodies, less than or equal to 25.0 signal units. Parameter match increased with progression of clinical status. These parameters were applied in a brief follow-up study of 34 HIV-infected asymptomatic individuals who developed AIDS-related complex or AIDS. Initial specimens showed a stronger correlation than our population data base had predicted, suggesting that the parameters have prognostic value. Densitometric analysis of antibody responses on Western blot assays of single or serial specimens should prove useful to physicians in staging and monitoring HIV-infected individuals and in predicting which individuals will progress to AIDS.
Detection of Clostridium difficile toxin in various tissue culture monolayers
Authors: Maniar A C , Williams T W , Hammond G W
Published: January 2020

Thirty stool filtrates known to contain Clostridium difficile toxin based on previous testing on McCoy cells were tested for toxicity on primary African green monkey kidney (AGMK), McCoy, MRC-5, primary rhesus monkey kidney (RMK), and Vero cells. All 30 filtrates showed cytotoxic effect at greater than or equal to 1:100 dilution on McCoy and Vero cells. A total of 22 filtrates were positive on MRC-5 monolayers, while only 16 and 10 filtrates showed positive cytotoxic effect on AGMK and RMK cells, respectively. Another 630 stool specimens were tested on McCoy and Vero cells only. Of these stool filtrates, 70 were positive and 560 were negative with both cell lines, which thus gave 100% agreement. Vero cells can be used interchangeably with McCoy cells for the detection of C. difficile toxin in stool filtrates.
Porphyrin test as an alternative to benzidine test for detecting cytochromes in catalase-negative gram-positive cocci
Authors: Wong J D
Published: January 2020

A total of 66 strains of gram-positive cocci, including 21 catalase-negative members of the family Streptococcaceae and strains of Stomatococcus mucilaginosus, were investigated for the ability to produce porphobilinogen and porphyrin from delta-aminolevulinic acid as an alternative to the benzidine test for detecting the presence of cytochromes. Production of porphobilinogen correlated 100% with membership in the family Micrococcaceae.
Successive bacteremias with "Campylobacter cinaedi" and "Campylobacter fennelliae" in a bisexual male
Authors: Ng V L , Hadley W K , Fennell C L , Flores B M , Stamm W E
Published: January 2020

A bisexual human immunodeficiency virus-seropositive male had successive bacteremias with "Campylobacter cinaedi" and "Campylobacter fennelliae." Because final identification of both isolates was not completed until 1 month after the last admission of the patient, a novel and nonstandardized antimicrobial susceptibility testing method was useful in guiding timely antimicrobial therapy.
Improved methods for isolation and enumeration of Malassezia furfur from human skin
Authors: Leeming J P , Notman F H
Published: January 2020

A medium for the isolation and enumeration of Malassezia furfur is described. Incubation at 34 degrees C yielded geometric mean counts (in CFU per square centimeter) of 2.6 X 10(3) on the forehead, 8.5 X 10(2) on the cheek, and 9.6 X 10(3) on the back. These counts compared favorably with microscopic counts and greatly exceeded those obtained with previously described media.
Comparison of two transport systems for recovery of aerobic and anaerobic bacteria from abscesses
Authors: Brook I
Published: January 2020

An evaluation of two commercially available swab transport systems, Port-A-Cul (PAC; BBL Microbiology Systems, Cockeysville, Md.) and Anaerobic Specimen Collector (ASC; Becton Dickinson Vacutainer Systems, Rutherford, N.J.), in the recovery of organisms from clinical specimens was done. Fifteen abscesses were drained, and swabs of their contents were placed in the transport systems until they were inoculated for detection of aerobic and anaerobic bacteria. The swabs were plated immediately after collection and after delays of 4, 24, 48, and 72 h. A total of 70 bacterial isolates, 47 anaerobes and 23 aerobes, were recovered from specimens inoculated immediately after collection. The survival of anaerobic bacteria was better in the PAC system than in the ASC system. This was evident as the length of delay in cultivation was extended. At 4 h, 46 anaerobic isolates were recovered in the PAC system, compared with 39 in the ASC system (P less than 0.1). At 24 h, 45 isolates were recovered in the PAC system and 26 isolates were recovered in ASC (P less than 0.001); at 48 h, 40 were recovered in PAC and 15 were recovered in ASC; and at 72 h, 32 were recovered in PAC and 6 were recovered in ASC. There were no differences between the systems in the recovery of aerobic bacteria. These data demonstrate the usefulness of the PAC system in the recovery of anaerobes and the need for quality control of all transport systems for anaerobic bacteria.
Evaluation of coagulase-negative staphylococci for ability to produce toxic shock syndrome toxin 1
Authors: Kreiswirth B N , Schlievert P M , Novick R P
Published: January 2020

A large and diverse group of coagulase-negative staphylococci were assayed for the ability to produce toxic shock syndrome toxin 1 (TSST-1) by immunological reactivity, and whole-cell DNAs from 33 of these strains were hybridized with a TSST-1-specific gene probe. None of the strains tested, including isolates that have been reported as TSST-1+, produced the exotoxin, and no DNA homology was found with the gene probe.
Improved toluidine blue-DNA agar for detection of DNA hydrolysis by campylobacters
Authors: Lior H , Patel A
Published: January 2020

Our improved toluidine blue-DNA agar was compared with methyl green DNase test agar for the detection of DNA hydrolysis by campylobacters. The toluidine blue-DNA agar gave clear-cut positive and negative reactions with campylobacter strains belonging to several species.
Synergistic hemolysis associated with coagulase-negative staphylococci isolated from bovine mammary glands
Authors: Watts J L , Owens W E
Published: January 2020

A total of 353 coagulase-negative staphylococcus (CNS) isolates from infected bovine mammary glands were tested for cytolysin production by using the synergistic hemolysin assay (SHA). Overall, 34.6% of CNS isolates were SHA positive. Human-associated, coagulase-negative staphylococcal species contained the greatest number of SHA-positive strains. Milk leukocyte levels expressed as somatic cell counts (SCC) were elevated with SHA-positive Staphylococcus epidermidis, Staphylococcus hominis, and Staphylococcus warneri strains. Elevated SCC levels were associated with strains of Staphylococcus hyicus and Staphylococcus chromogenes. However, no difference in SCC levels was observed between SHA-positive and SHA-negative strains. Results indicated that the SHA was a sensitive test for the detection of cytolysin-producing CNS.
Evaluation of MUREX SUDS Toxo test
Authors: Moyer N P , Hudson J D , Hausler W J
Published: January 2020

The SUDS Toxo test (MUREX Corp., Norcross, Ga.) was compared with the indirect hemagglutination test (IHA) and the indirect fluorescent-antibody test (IFA) by examining 404 serum specimens, including 64 (15.8%) specimens with IFA titers of greater than or equal to 1:2. When SUDS was compared with IHA, sensitivity (96.4%), specificity (97.9%), and negative predictive value (99.4%) indicated that there were similar reactivities between the two tests. When an IFA titer of greater than or equal to 1:16 was considered significant and IHA and SUDS were compared with IFA, IHA was slightly less sensitive but had a higher positive predictive value than did SUDS; however, there was no statistical difference between the tests. When SUDS was compared with IFA, in which a titer of greater than or equal to 1:16 was considered significant, the high negative predictive value (100%), excellent sensitivity (100%) and specificity (98.3%), and ease of performance made SUDS an attractive alternative to IHA for screening single serum specimens for toxoplasmosis.
Antigenic analysis of fimbrial proteins from Moraxella bovis
Authors: Moore L J , Rutter J M
Published: January 2020

Fimbrial proteins were extracted from 15 isolates of Moraxella bovis, and antisera to each of the preparations were raised in rabbits. The antigenic relationships of the fimbriae were investigated by an enzyme-linked immunosorbent assay, tandem crossed immunoelectrophoresis, and a slide agglutination test. With all three methods there was a similar pattern of antigenic cross-reactivity among the fimbriae. The 15 isolates, together with 23 additional isolates, could be grouped into seven fimbrial serogroups.
Evaluation of Quantum II microbiology system for identification of gram-negative bacteria of veterinary origin
Authors: Jones R L , Adney W S , Davis M A , Vonbyren H , Thompson G
Published: January 2020

The ability of a rapid, semiautomated bacterial identification system, the Quantum II microbiology system (Abbott Laboratories, Irving, Tex.), to accurately identify gram-negative bacteria from veterinary sources was evaluated. A total of 378 isolates were tested, including 298 organisms in the family Enterobacteriaceae and strains representing Acinetobacter sp., Aeromonas sp., Flavobacterium sp., Pasteurella multocida, Plesiomonas sp., and Pseudomonas spp. Of these isolates, 333 (88.1%) were correctly identified, 20 (5.3%) were not identified, 10 (2.6%) were incorrectly identified at the genus level, and 15 (4.0%) were incorrectly identified at the species level. The Quantum II system correctly identified 268 (89.9%) of the isolates of Enterobacteriaceae and 65 (81.3%) of the nonenteric isolates. P. multocida was not identified correctly, and some nonenteric gram-negative bacteria of clinical significance in veterinary medicine are not included in the data base. The Quantum II system provided an accurate identification system for isolates of Enterobacteriaceae but had limited usefulness for the identification of other gram-negative bacteria of clinical significance in veterinary medicine.
Genetic and antigenic differences of serologically indistinguishable leptospires of serovar hardjo
Authors: LeFebvre R B , Thiermann A B , Foley J
Published: January 2020

Pathogenic leptospires of serovar hardjo isolated from North American cattle were compared genetically and antigenically to reference strain hardjoprajitno of the Sejroe serogroup. Guanine-plus-cytosine (G+C) contents were determined for the genomes, and microscopic agglutination, Western blotting (immunoblotting), and immunoprecipitation were used to characterize antigenic relatedness. Major differences were demonstrated between the isolates and the reference strain. The G+C content of the reference strain was calculated to be approximately 34 +/- 1%, and those of the isolates were calculated at 39 +/- 1%. Antigenic differences between the isolates and the reference strain were identified by using rabbit immune serum raised against a hardjo isolate exhaustively adsorbed with hardjoprajitno whole and sonicated cells. Western blot analysis and immunoprecipitation using this adsorbed serum revealed antigens apparently unique for the hardjo isolates. Microscopic agglutination with the adsorbed rabbit serum did not agglutinate hardjoprajitno when diluted 1:2 but agglutinated bovine isolates to a 1:32 dilution. Bovine antiserum raised against the isolates was also used to identify antigens by immunoprecipitation.
Improved medium for antimicrobial susceptibility testing of Haemophilus influenzae
Authors: Jorgensen J H , Redding J S , Maher L A , Howell A W
Published: January 2020

The need for complex growth media has complicated routine susceptibility testing of Haemophilus influenzae because of antagonism of certain antimicrobial agents by the medium or because of difficulties in interpretation of growth endpoints. Haemophilus test medium (HTM) is a simple, transparent medium for broth- or agar-based tests with H. influenzae. HTM incorporates Mueller-Hinton medium with additions of 15 micrograms of hematin per ml, 15 micrograms of NAD per ml, and 5 mg of yeast extract per ml as growth-promoting additives. Agar or broth microdilution MICs of 10 antimicrobial agents for a collection of 179 H. influenzae isolates determined by using HTM compared favorably with MICs determined by the conventional agar or broth dilution methods recommended by the National Committee for Clinical Laboratory Standards. Disk diffusion tests performed with HTM allowed accurate categorization of susceptible and resistant strains and were easier to interpret than tests performed with Mueller-Hinton chocolate agar. A particular advantage of HTM was the reliability of broth- or agar-based test results with trimethoprim-sulfamethoxazole. The results of the study suggest modification of current National Committee for Clinical Laboratory Standards MIC-interpretive criteria for H. influenzae with amoxicillin-clavulanate, chloramphenicol, and trimethoprim-sulfamethoxazole. Error rate-bounded analysis of MICs and disk diffusion zone sizes also suggest modified zone-interpretive criteria for ampicillin, amoxicillin-clavulanate, chloramphenicol, and tetracycline with HTM or conventional media. Interpretive zone sizes are newly proposed for cefaclor and rifampin disk diffusion tests.
Indirect enzyme-linked immunosorbent assay for measurement of human immunoglobulins E and G to purified cow's milk proteins: application in diagnosis of cow's milk allergy
Authors: Campbell D E , Ngamphaiboon J , Clark M M , Harris M C , Kolski G B , Douglas S D
Published: January 2020

An indirect double-antibody enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of human immunoglobulin E (IgE) and IgG to the cow's milk proteins (CMP) alpha-casein, alpha-lactalbumin, and beta-lactoglobulin. Human serum albumin was used as the negative-antigen control. Rabbit anti-human IgE or IgG served as the primary antibody, and horseradish peroxidase-conjugated swine anti-rabbit immunoglobulin served as the secondary antibody. Positive control sera were obtained from patients with well-documented histories of cow's milk allergy, while negative control sera were obtained from cord bloods of healthy full-term infants and from normal adult volunteers without known milk allergy. Test sera were obtained from 41 children (ages, 3 months to 13 years; average age, 2.6 years) with suspected cow's milk allergy and clinical manifestations that included wheezing, rhinitis, atopic dermatitis, urticaria, or gastrointestinal disturbances. The patients were simultaneously evaluated by prick skin testing with scratch test antigen to whole CMP. Although only 13 (32%) of the 41 patients were positive by the prick skin test, 25 (61%) were positive by the IgE ELISA. Of the 25 IgE ELISA-positive patients, 20 were also positive by the IgG ELISA. There was concordance of positive results between skin testing and the IgE ELISA in only 9 patients (22%), and there was concordance of negative results in 12 patients (29%). Discordant results were observed in 20 patients (49%). These results indicate that the ELISA is more sensitive than prick skin testing in the identification of individuals with elevated levels of IgE to CMP.
Legionella birminghamensis sp. nov. isolated from a cardiac transplant recipient
Authors: Wilkinson H W , Thacker W L , Benson R F , Polt S S , Brookings E , Mayberry W R , Brenner D J , Gilley R G , Kirklin J K
Published: January 2020

A Legionella-like organism, strain 1407-AL-H, was isolated from a transbronchial lung biopsy specimen from a cardiac transplant recipient undergoing immunosuppressive therapy. The strain grew on buffered charcoal-yeast extract agar (BCYE) but not on BCYE in the absence of cysteine, and it showed gas-liquid chromatographic fatty acid profiles that were predominantly branch chained. Strain 1407-AL-H was antigenically distinct in slide agglutination tests from the 23 Legionella species and 39 serogroups previously described. DNA hybridization studies placed it in a new Legionella species, Legionella birminghamensis (ATCC 43702).
High-performance liquid chromatography analysis of mycolic acids as an aid in laboratory identification of Rhodococcus and Nocardia species
Authors: Butler W R , Kilburn J O , Kubica G P
Published: January 2020

High-performance liquid chromatography analysis of the p-bromophenacyl esters of mycolic acids from whole organisms gave chromatographic patterns that were useful in differentiation of Rhodococcus and Nocardia species. Rhodococcus equi, R. erythropolis, and R. rhodochrous contained more-polar mycolic acids and were easily separated from the less-polar mycolic acid-containing species of R. sputi, R. bronchialis, R. corallinus, R. rubropertinctus, and R. terrae. The less-polar mycolic acid-containing Rhodococcus species showed chromatographic patterns that partially overlapped (in elution times) the patterns of Nocardia asteroides, N. otitidiscaviarum, and N. brasiliensis, but the larger number of peaks in the last species made separation between the genera possible. Distinct chromatographic patterns were found for most species, except for R. equi strains that showed two different patterns. Strains of R. rubropertinctus and R. terrae appeared identical. N. asteroides and N. otitidiscaviarum showed similar mycolic acid patterns.
Escherichia coli associated with childhood diarrheas
Authors: Yam W C , Lung M L , Yeung C Y , Tam J S , Ng M H
Published: January 2020

We studied 2,246 episodes of childhood diarrhea over a 2-year period in a general hospital serving a population of about one million on the island of Hong Kong. Rotavirus (24%) and nontyphoid salmonellas (23%) were the most common causal agents, followed by Campylobacter sp. (9%). Rotavirus occurred largely during winter, whereas salmonellas and Campylobacter sp. occurred more commonly during summer and autumn, respectively. Enterotoxigenic Escherichia coli was rarely isolated (1%), and the isolates were clonally diverse. A small percentage (2.8%) of E. coli had serological specificities commonly associated with enteropathogenic E. coli, but only two of the isolates were also positive for HEp-2 adhesive factor. Pure or heavy and predominant growth of E. coli was obtained in repeated stool cultures of 432 (19%) of these episodes, which did not yield any of the above-mentioned pathogens. Although associated with diarrhea, these E. coli isolates possess neither the pathogenic attributes of enterotoxigenic E. coli nor the properties commonly associated with enteropathogenic E. coli. Enteroinvasive E. coli and enterohemorrhagic E. coli were considered unlikely causes on clinical grounds.
Legionella micdadei and Legionella dumoffii monoclonal antibodies for laboratory diagnosis of Legionella infections
Authors: Cercenado E , Edelstein P H , Gosting L H , Sturge J C
Published: January 2020

Two different monoclonal antibodies directed against Legionella micdadei and L. dumoffii (Genetic Systems Corp., Seattle, Wash.) were evaluated for their specificity and ability to detect L. micdadei and L. dumoffii in human and animal clinical samples and bacterial isolates in an indirect immunofluorescence assay. All three frozen sputum samples and all three Formalin-fixed sputum and liver samples from patients with culture-documented L. micdadei pneumonia were positive when tested with the L. micdadei monoclonal antibody. A Formalin-preserved lung sample from a patient with culture-documented L. dumoffii pneumonia was positive with its homologous monoclonal antibody. No cross-staining reactions were found with either monoclonal antibody on any of 25 human sputum samples tested from patients without Legionella infections. A total of 66 Legionella strains and 56 non-Legionella strains including 22 Pseudomonas strains and 34 other bacterial strains were studied. No cross-staining reactions were found except in Staphylococcus aureus Cowan 1 ATCC 12598. The lower limit of detection in seeded sputum samples was about 7 X 10(4) cells per ml for both monoclonal antibodies. Lung and tracheal lavage specimens from L. micdadei- or L. dumoffii-infected guinea pigs showed specific staining only with their respective monoclonal antibodies. The monoclonal antibodies stained homologous bacteria slightly less intensely than did the polyclonal antisera, but the signal-to-noise ratio was considerably higher for the monoclonal antibodies. No differences in sensitivity of staining of clinical specimens or bacterial isolates were noted between the monoclonal antibodies and the polyclonal reagents for L. micdadei and L. dumoffii (Centers for Disease Control, Atlanta, Ga., and BioDx, Denville, N.J. These monoclonal antibodies ae sensitive and specific, making them good candidates for laboratory diagnostic purposes.
Hyperimmune serum from rabbits immunized with potassium thiocyanate extract of Pasteurella multocida protects against homologous challenge
Authors: Lu Y S , Pakes S P , Massey L
Published: January 2020

Hyperimmune rabbit sera directed to the KSCN extract of 3:A Pasteurella multocida were characterized by enzyme-linked immunosorbent assay (ELISA), presolubilized cell radioimmunoprecipitation, and immunoblotting analysis. The results showed that the hyperimmune serum had a very high titer of immunoglobulin G ELISA antibody and a negligible immunoglobulin A ELISA antibody, precipitated 10 different outer membrane protein antigens by radioimmunoprecipitation, and reacted to 10 different membrane vesicle antigens of P. multocida by immunoblotting analysis. The hyperimmune rabbit sera were also evaluated for protective efficacy against experimental rabbit pasteurellosis by homologous challenge. Thirty-six rabbits were divided into four groups. Group 1, 2, and 3 rabbits were inoculated intranasally with hyperimmune rabbit serum, phosphate-buffered saline, or normal rabbit serum, respectively, at 24 h prior to and 24, 48, and 72 h after intranasal challenge with the virulent homologous P. multocida strain. Group 4 rabbits were inoculated with normal rabbit serum without challenge. Necropsies of surviving rabbits were performed 2 weeks postinfection. The mortality rates for groups 1 through 4 were 25% (3 of 12), 67% (8 of 12), 75% (6 of 8), and 0% (0 of 4), respectively. The prevalence and severity of pneumonia were significantly lower in the hyperimmune serum-treated rabbits. The prevalence of P. multocida colonization in lungs was significantly lower in group 1 rabbits, and the geometric mean CFU of P. multocida in lungs was 59,166-fold less in group 1 rabbits than in group 3 rabbits. The geometric mean CFU of P. multocida in nasal cavities of group 1 rabbits was significantly lower than that of group 3 rabbits. All challenged rabbits (groups 1,2, and 3) had elevated nasal immunoglobulin A and pulmonary (lung lavage) immunoglobulin A antibody levels at necropsy (day 14 postinfection). Similarly, all challenged rabbits had elevated levels of ELISA immunoglobulin G antibody in serum at day 14 but not at day 7 postinfection, indicating that rabbits receiving hyperimmune serum can mount a specific humoral immune response against the homologous challenge P. multocida organisms. We concluded that hyperimmune serum directed to the KSCN extract of 3:A P. multocida provides significant protection against homologous challenge in rabbits.
Rapid diagnosis of cholera by coagglutination test using 4-h fecal enrichment cultures
Authors: Rahman M , Sack D A , Mahmood S , Hossain A
Published: January 2020

A simple, rapid, and reliable method to detect Vibrio cholerae in fecal specimens would assist in the management of cases of severe diarrhea, especially since most such cases occur in areas with minimal laboratory facilities. A coagglutination test was used to detect V. cholerae antigen in bile-peptone broth incubated with feces. In the technique, Staphylococcus aureus Cowan 1 coated with anti-V. cholerae O1 antiserum was tested with cultures incubated for 4 h. When 165 specimens were tested, the sensitivity, specificity, and accuracy of the test, compared with standard culture methods, were 97, 99, and 98%, respectively. These promising results were better than those of dark-field microscopy using the same specimens, and the test was logistically easy to perform. The coagglutination test using enrichment broth culture of feces is a simple and rapid method which may be used to confirm a diagnosis of cholera.