bluuebunny/crossref_metadata_2025 huggingface dataset:
Resistance to serum bactericidal activity distinguishes Brazilian purpuric fever (BPF) case strains of Haemophilus influenzae biogroup aegyptius (H. aegyptius) from non-BPF strains. Brazilian Purpuric Fever Study Group
Authors: Porto M H , Noel G J , Edelson P J
Published: January 2020

We studied the ability of normal human serum to lyse H. influenzae biogroup aegyptius (H. aegyptius) isolates recovered from patients with Brazilian purpuric fever (BPF clone) or non-BPF clone strains. BPF clone isolates, although similar to non-BPF clone isolates with regard to the ability to fix C3 to their surfaces, could be distinguished from non-BPF clone strains by their resistance to lysis in vitro following incubation with normal adult human serum.
Comparison of colorimetric, fluorescent, and enzymatic amplification substrate systems in an enzyme immunoassay for detection of DNA-RNA hybrids
Authors: Coutlee F , Viscidi R P , Yolken R H
Published: January 2020

The monoclonal antibody solution hybridization assay is a novel enzyme immunoassay for detection of RNA with a biotinylated DNA probe. To increase the sensitivity of this test, a fluorescent substrate and an enzymatic amplification cycling system were compared with a conventional colorigenic substrate for alkaline phosphatase. The fluorescent, cycling, and colorigenic substrates detected, respectively, 10, 10, and 100 amol of unbound alkaline phosphatase in 2 h. With a prolonged incubation period of 16.6 h, the conventional substrate measured 10 amol of the enzyme. In the immunoassay for RNA detection, the fluorescence and cycling assays were faster than that using the colorigenic substrate and reached an endpoint sensitivity of 3.2 pg/ml (0.16 pg per assay) of cRNA. However, longer incubation periods (16.6 h) for optimal generation of the colorigenic product led to a comparable level of sensitivity for the conventional substrate.
Serological classification of Xanthomonas maltophilia (Pseudomonas maltophilia) based on heat-stable O antigens
Authors: Schable B , Rhoden D L , Hugh R , Weaver R E , Khardori N , Smith P B , Bodey G P , Anderson R L
Published: January 2020

Twenty-six serotypes of Xanthomonas maltophilia were defined by using 15 antisera described by Hugh and Ryschenkow (R. Hugh and E. Ryschenkow, J. Gen. Microbiol. 26:123-132, 1961) and 11 new antisera. The antisera were prepared by immunizing rabbits with bacterial strains heated at 100 degrees C for 2 h. Twelve antisera required adsorptions with cross-reacting heterologous immunizing strains. We tested 275 clinical and environmental strains of X. maltophilia with 26 antisera by the slide agglutination technique. A total of 259 (94.2%) strains were typeable, with 137 (49.8%) strains agglutinating in three antisera.
Rapid detection of herpes simplex virus with fluorescein-labeled Helix pomatia lectin
Authors: Slifkin M , Cumbie R
Published: January 2020

The use of fluorescein-conjugated Helix pomatia lectin was shown to be as effective as fluorescein-conjugated monoclonal antibody reagents for the detection and differentiation of herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2) in MRC-5 cell culture. Cells infected with HSV-1 generally displayed a pattern of nongranular or diffuse fluorescence, while cells infected with HSV-2 were identified by the production of fluorescent grains and flecks. This unique nonimmunological reagent, when used in combination with low-speed centrifugation, provides a remarkably specific, sensitive, rapid, and cost-effective means to detect HSV-infected MRC-5 or BHK-21 cells as early as 20 h postinoculation. In contrast to the immunofluorescence method, the serotypes of HSV can be differentiated with only one fluorescein-H. pomatia reagent in MRC-5 cell cultures.
Prospective study of Campylobacter jejuni infection in Chilean infants evaluated by culture and serology
Authors: Figueroa G , Galeno H , Troncoso M , Toledo S , Soto V
Published: January 2020

A prospective study of Campylobacter jejuni infection was performed during a 6-month period in a cohort of 198 Chilean infants. Surveillance was based on biweekly home visits by a team of trained nurses. C. jejuni colonization was studied by culturing stool samples from diarrheal episodes and paired asymptomatic controls and by performing an enzyme-linked immunosorbent assay developed to measure C. jejuni outer membrane protein polyvalent antibodies in a representative group of infants. C. jejuni was isolated from 30 of 299 diarrheal episodes (10%) and from 17 of 304 samples from asymptomatic infants (6%). Significantly higher (P less than 0.05) C. jejuni illness/infection ratios were found in infants less than 9 months old. Polyvalent antibodies to C. jejuni were found in 27 of 89 serum samples (30%). The lowest prevalence (4%) was observed among infants less than 9 months old, and the highest was found in patients older than 15 months (63%; P less than 0.01). Data obtained by analysis of paired serum samples revealed a significant increase in the number of seropositive individuals, from 8% on admission to 50% at the end of the protocol (P less than 0.001). The change in the immune status of these infants was associated with symptomatic infection in 9 of 18 cases. The antibody rise found in the remaining nine infants suggested the presence of asymptomatic C. jejuni infections. This inference was documented by the isolation of C. jejuni from stools of two of these infants when the infants were randomly studied while asymptomatic. This study points out the impact of C. jejuni-associated diarrheal episodes and the high frequency with which asymptomatic infections elicit specific antibodies in infants living in areas such as Chile.
Time to positivity for detection of bacteremia in neonates
Authors: Kurlat I , Stoll B J , McGowan J E
Published: January 2020

The blood culture results of all samples obtained from newborns at Grady Memorial Hospital, Atlanta, Ga., during a 6-month period were analyzed to determine the time required for a blood culture to become positive, the time at which a culture could safely be considered negative, and the spectrum of isolated organisms. During the study period, 1,248 samples were submitted from all nurseries and processed by an automated detection instrument (BACTEC NR660). Of the 1,248 samples, 98 (7.8%) were positive by the end of a 7-day processing period; 29 of the 98 were classified as definite bacterial pathogens, 52 were classified as possible bacterial pathogens, 9 were classified as yeasts, and 8 were classified as contaminants. Virtually all organisms (28 of 29) categorized as definite pathogens were identified by day 2 of processing, and all were identified by day 4. All isolates of group B streptococcus, Escherichia coli, Klebsiella species, and Staphylococcus aureus were identified by day 2. Of all positive blood cultures, 79% were identified by day 2, 88% were identified by day 3, and 94% were identified by day 4. Of the 21 isolates identified after day 2, the only definite pathogen was from a sick baby in the intensive care unit. From among the 870 term low-risk newborns cultured because of maternal risk factors, only four possible pathogens were identified after day 2. The positive and negative predictive values of blood culture at days 2 and 4 were 92 and 99%, respectively.We conclude that, in our institution, (i) a 2-day processing period is sufficient to detect positive blood cultures in the asymptomatic term infant, (ii) a 4-day processing period will detect virtually all clinically important infections, and (iii) clinical yield from continuing blood culture processing beyond 4 days does not justify the time and cost involved.
Laboratory investigation of hospital outbreak caused by two different multiresistant Acinetobacter calcoaceticus subsp. anitratus strains
Authors: Vila J , Almela M , Jimenez de Anta M T
Published: January 2020

During a 7-month period, from December 1986 to June 1987, multiresistant strains of Acinetobacter calcoaceticus subsp. anitratus were isolated from 25 patients in a respiratory intensive care unit. The biochemical characteristics defined two groups of strains, group 1 (14 strains) and group 2 (11 strains). Both groups had the same biochemical characteristics, but group 2 strains could assimilate adipate and phenyl acetate. Moreover, of 16 antibiotics tested only netilmicin and imipenem had some inhibitory activity for group 1 strains; group 2 strains were susceptible to mezlocillin, piperacillin, and ticarcillin. Plasmid profiles of the groups were also different. The results of a laboratory investigation (biochemical characteristics, antibiotic susceptibility, and plasmid isolation) identified two different A. calcoaceticus subsp. anitratus strains as the causes of the outbreak.
Detection of antigens in urine during acute toxoplasmosis
Authors: Huskinson J , Stepick-Biek P , Remington J S
Published: January 2020

Toxoplasma antigens were detected in sera and urine of mice acutely infected with Toxoplasma gondii. The concentrations of antigens in the urine samples measured by enzyme-linked immunosorbent assay were similar to those detected in the sera of the corresponding mice. The major antigens were not dialyzable and were largely destroyed by treatment with trichloroacetic acid and heat (100 degrees C for 1 h). Toxoplasma antigens were demonstrable on Western blots (immunoblots) of the urine samples.
Genetic typing in a cluster of Legionella pneumophila infections
Authors: van Ketel R J , de Wever B
Published: January 2020

Legionella pneumophila strains isolated from six patients, three air-conditioning- and cooling tower-derived strains, and three hot water supply-derived strains were analyzed by three genetic typing methods. The results of the whole-cell DNA restriction endonuclease analysis and the restriction patterns based on genes coding for rRNA correlated with each other and demonstrated that the patient isolates were indistinguishable from the air-conditioning- and cooling tower-derived isolates but differed markedly from the hot water supply-derived isolates. The patient and air-conditioning- and cooling tower-derived strains contained plasmids of the same molecular weight; the hot water supply-derived strains were plasmidless. These results indicated that the cooling tower or the air-conditioning system was the environmental source for the examined cluster of Legionnaires disease strains.
Use of a DNA probe to detect Salmonella typhi in the blood of patients with typhoid fever
Authors: Rubin F A , McWhirter P D , Punjabi N H , Lane E , Sudarmono P , Pulungsih S P , Lesmana M , Kumala S , Kopecko D J , Hoffman S L
Published: January 2020

A DNA probe was used to detect Salmonella typhi from blood samples from 14 of 33 patients with culture-confirmed typhoid fever, using the equivalent of 2.5 ml of blood. In contrast, S. typhi was detected in 17 of the same 33 patients by culture of 8 ml of blood. The probe hybridized to blood samples of 4 of 47 patients from whom S. typhi was not isolated.
Yersinia enterocolitica in donor blood: a case report and review
Authors: Jacobs J , Jamaer D , Vandeven J , Wouters M , Vermylen C , Vandepitte J
Published: January 2020

Routine sterility control of a unit of leukocyte-depleted erythrocyte concentrate yielded growth of Yersinia enterocolitica serotype O:3. Plasma of the donor showed a high titer of agglutinins against the homologous organism. Although the donor was apparently well at the time of donation, he had a history of protracted terminal ileitis treated by surgery. The recipient of the contaminated blood was taking broad-spectrum antibiotics and did not experience any adverse effect. Fourteen other cases of transfusion-associated yersiniosis have been reported.
Liver abscess caused by Clostridium bifermentans following blunt abdominal trauma
Authors: Nachman S , Kaul A , Li K I , Slim M S , San Filippo J A , Van Horn K
Published: January 2020

A case of hepatic abscess and subsequent septicemia caused by Clostridium bifermentans is described. The abscess manifested itself on the third day after blunt trauma to the torso. The patient had nausea, vomiting, fever, evidence of hepatic dysfunction, and subphrenic gas. This case illustrates the association of hepatic abscess and blunt trauma to the torso.
CDC group HB-5 as a cause of genitourinary infections in adults
Authors: Baddour L M , Gelfand M S , Weaver R E , Woods T C , Altwegg M , Mayer L W , Kelley R A , Brenner D J
Published: January 2020

Isolates from five patients with genitourinary infections diagnosed over a 7-week period were identified as CDC group HB-5. The organisms caused clinical presentations of pelvic inflammatory disease in three women and urethritis in the only male in the series. The remaining patient received surgical treatment for a Bartholin gland abscess. Since the clinical and temporal presentations were remarkable and the questions of sexual mode of transmission and strain relatedness were of concern, the five strains were examined further by multilocus enzyme electrophoresis and ribosomal DNA typing. Overall, the five clinical isolates were more closely related to each other than to the control strains. This is the first report describing genitourinary infections caused by group HB-5. While the pathophysiology of group HB-5 infections remains to be fully elucidated, the possibility that this organism may be sexually transmitted deserves further study.
Comparison of DNA probe, monoclonal antibody enzyme immunoassay, and cell culture for the detection of Chlamydia trachomatis
Authors: LeBar W , Herschman B , Jemal C , Pierzchala J
Published: January 2020

A total of 201 endocervical specimens were obtained from patients with a clinical or epidemiological history suggestive of chlamydial infection. These specimens were tested by DNA probe (Gen-Probe, San Diego, Calif.) and the IDEIA III (Boots-Celltech, Berkshire, United Kingdom) monoclonal antibody enzyme immunoassay and compared with cell culture for detection of Chlamydia trachomatis. Discrepancies between cell culture and antigen detection methods were resolved by direct fluorescent-antibody testing. In a population with a 17.4% prevalence, the sensitivities and specificities of these assays were 82.8 and 99.4%, respectively, for the DNA probe assay and 97.1 and 98.1%, respectively, for the IDEIA III.
Detection of influenza C virus by using an in situ esterase assay
Authors: Wagaman P C , Spence H A , O'Callaghan R J
Published: January 2020

A variety of chemically defined compounds were tested to characterize the substrate specificity of the influenza C virus esterase and to determine whether a substrate could be found that would be useful in an assay to detect the virus. Two new substrates, alpha-naphthyl acetate and alpha-naphthyl propionate, were identified; alpha-naphthyl acetate was employed to develop an assay specific for influenza type C virus in MDCK cells. The assay was sufficiently sensitive to detect esterase activity in a single cell and distinguished influenza C virus infections from those of types A and B viruses. Infected cells could be detected as early as 8 h postinfection, with maximal enzyme detection occurring at 24 h. Assay of influenza C virus in the chorioallantoic or amniotic fluid of infected eggs was performed by applying fluids directly onto nitrocellulose strips and then incubating with alpha-naphthyl acetate. Both the cellular and nitrocellulose-bound assays are rapid, inexpensive, and easy to perform, offering advantages for use in clinical laboratories.
Characterization of the Aeromonas hydrophila group isolated from retail foods of animal origin
Authors: Palumbo S A , Bencivengo M M , Del Corral F , Williams A C , Buchanan R L
Published: January 2020

During a recent survey of retail fresh foods of animal origin (fish and seafood, raw milk, poultry, and red meats) for organisms of the Aeromonas hydrophila group, we isolated representative strains from the various foods. In this study, we sought to characterize these isolates for biochemical properties and virulence-associated factors and to compare the food isolates with clinical isolates. We identified all food and clinical isolates as A. hydrophila and found that all isolates were typical in their biochemical reactions. Examination of the isolates for various virulence-associated factors indicated that most food and clinical isolates were serum resistant, beta-hemolytic, cytotoxin positive (against Y1 adrenal cells), hemagglutinin positive, Congo red positive, elastase positive, and staphylolysin positive. Mouse 50% lethal doses were log10 8 to 9 CFU for most isolates. All isolates had biotypes identical to those of enterotoxin-positive strains. The public health significance of these organisms in foods is not known at present, although their widespread occurrence and ability to grow competitively in foods kept at 5 degrees C represents a potential hazard.
Noninfectious rotavirus (strain RRV) induces an immune response in mice which protects against rotavirus challenge
Authors: Offit P A , Dudzik K I
Published: January 2020

We found that female adult mice parenterally inoculated with noninfectious rotavirus (simian strain RRV) developed virus-specific neutralizing antibodies in the serum; newborn mice from these dams were protected against RRV-induced gastroenteritis. In addition, mice parenterally inoculated with noninfectious RRV developed virus-specific cytotoxic T-lymphocyte precursors in the spleen. Replication of rotavirus in intestinal epithelial cells was apparently not required to induce rotavirus-neutralizing antibodies or rotavirus-specific cytotoxic T lymphocytes. Parenteral immunization of infants and young children with noninfectious rotaviruses may induce an immune response which protects against rotavirus challenge.
Enzyme-linked immunosorbent assay for determination of antibodies to the envelope glycoprotein of rabies virus
Authors: Grassi M , Wandeler A I , Peterhans E
Published: January 2020

The envelope glycoprotein G of rabies virus induces neutralizing antibodies, which are important in protection against rabies. This protein was solubilized from purified virus and isolated by differential and sucrose density gradient centrifugation followed by high-performance liquid chromatography. Conditions for solubilization and purification of G were optimized by using immunoblotting and enzyme-linked immunosorbent assay techniques. The reaction with conventional antisera and monoclonal antibodies indicated that purified G protein was essentially devoid of internal viral proteins. Microdilution plates were coated with purified G protein, and sera from humans vaccinated against rabies were tested for the presence of antibodies. Results were compared with those of the rapid fluorescent focus inhibition assay, which is the standard neutralization assay for antibodies to rabies virus. The results of this comparison indicate that the enzyme-linked immunosorbent assay for G is a reliable and simple alternative to the neutralization test.
Competitive enzyme immunoassay for antibodies to a 43,000-molecular-weight Francisella tularensis outer membrane protein for the diagnosis of tularemia
Authors: Bevanger L , Maeland J A , Naess A I
Published: January 2020

Antibodies against a 43,000-molecular-weight Francisella tularensis outer membrane (OM) protein (43K protein) were measured in paired serum specimens from 23 patients with tularemia and matched against antibodies in sera from 25 patients with nontularemic infectious diseases and from 25 blood donors. Antibodies were measured by a competition enzyme-linked immunosorbent assay which tested the ability of human serum to compete with rabbit anti-43K protein antibodies for its binding to the F. tularensis OM coat. The sera from nontularemic patients and from blood donors, in dilutions of 1:16 and 1:64, respectively, showed no or very low levels of antibodies. All of the tularemia patients showed positive tests with the first, the second, or both of the serum specimens examined. For instance, with serum diluted 1:64, each of the serum specimens showed a sensitivity of 95.7% and a specificity of 96%. When used for antibody competition in Western blotting (immunoblotting), the rabbit anti-43K selectivity blocked the binding of human serum antibodies to the 43,000-molecular-weight protein. This protein was immunoaccessible in Formalin-killed F. tularensis. These data indicate an important role of the 43,000-molecular-weight OM protein in the immunobiology of tularemia and emphasize its potential usefulness as an antigen in serodiagnostic tests.
Evaluation of restriction endonuclease analysis as an epidemiologic typing system for Branhamella catarrhalis
Authors: Patterson J E , Patterson T F , Farrel P , Hierholzer W J , Zervos M J
Published: January 2020

Restriction endonuclease analysis (REA) was evaluated as an epidemiologic typing tool to distinguish Branhamella catarrhalis strains. Fourteen beta-lactamase-producing strains were collected over a 16-month period at a hospital where a nosocomial outbreak of this organism was previously documented by REA. REA produced 12 distinct patterns which correlated with epidemiologic data. Chromosomal REA appears to be a useful technique for distinguishing B. catarrhalis strains.
Occurrence of a common lipopolysaccharide antigen in standard and clinical strains of Pseudomonas aeruginosa
Authors: Lam M Y , McGroarty E J , Kropinski A M , MacDonald L A , Pedersen S S , Høiby N , Lam J S
Published: January 2020

The lipopolysaccharide (LPS) of Pseudomonas aeruginosa PAO1 contains two species of O polysaccharide termed A and B bands. The high-molecular-weight B-band LPS determines the O specificity of the bacterium, while the antigenically distinct A-band LPS consists of only shorter-chain polysaccharides. Seven hybridomas secreting A-band-specific monoclonal antibodies were produced and used to study the LPS of standard and clinical strains. Although A-band antibodies did not agglutinate any of the serotype strains presently in the International Antigenic Typing Scheme, Western immunoblots revealed that 11 of the 17 serotype strains possessed A-band LPS. In a group of 250 clinical isolates from patients with cystic fibrosis, 170 (68%) had A-band LPS on the basis of agglutination tests, but in silver-stained gels all were shown to be deficient in O-antigen-containing B band. Investigation of serial isolates from a single patient revealed a pattern of antigenic variation. During the course of the infection, serotypeable isolates became nontypeable, and the O antigen was replaced with A band as the major LPS antigen. These results suggest that A-band LPS may be the major LPS antigen in nontypeable clinical isolates and a common antigen among other P. aeruginosa strains.
Rapid method to detect shiga toxin and shiga-like toxin I based on binding to globotriosyl ceramide (Gb3), their natural receptor
Authors: Ashkenazi S , Cleary T G
Published: January 2020

Shiga toxin and the closely related Shiga-like toxins produced by Escherichia coli represent a group of very similar cytotoxins that may play an important role in diarrheal disease and hemolytic uremic syndrome. These toxins have the same biologic activities and according to recent studies also share the same binding receptor, globotriosyl ceramide (Gb3). They are currently detected, on the basis of their ability to damage several cell lines, by using expensive and tedious assays that require facilities for and experience with tissue cultures and are therefore most suitable for research laboratories. We have developed a rapid method to detect Shiga toxin and Shiga-like toxin I based on specific binding to their Gb3 natural receptor, which was coated onto microdilution plates. Bound toxin was then detected by enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies. The sensitivity of the Gb3 ELISA was 0.2 ng (2 ng/ml) of purified toxin. The assay was positive with sonic extracts of Shigella dysenteriae serotype 1 strain 6OR (a Shiga toxin producer), E. coli serotype O26:H11 strain H30, and E. coli serotype O157:H7 (both Shiga-like toxin I producers). The assay was very specific in that no cross-reactivity was noted with purified cholera toxin, E. coli heat-labile and heat-stable enterotoxins, and Clostridium difficile cytotoxin, or sonic extracts of other cytotoxin-producing organisms, such as other shigellae, pathogenic and nonpathogenic E. coli, Salmonella spp., Campylobacter spp., and Aeromonas spp. These results were in complete agreement with a [3H]thymidine-labeled HeLa cell cytotoxicity assay and with detection of the structural genes by DNA hybridization studies with a Shiga-like toxin I probe. Quantitative analysis showed a high correlation between Gb3 ELISA and HeLa cell assay when fractions obtained at various stages of toxin purification were examined by both methods (r = 0.99, P < 0.01). This rapid Gb3 ELISA is sensitive and specific and may be diagnostically useful in cytotoxin-related infections.
Detection of BK virus and JC virus in urine and brain tissue by the polymerase chain reaction
Authors: Arthur R R , Dagostin S , Shah K V
Published: January 2020

DNAs of the human polyomaviruses BK virus (BKV) and JC virus (JCV) were amplified by the polymerase chain reaction (PCR) by using a single pair of 20-base oligonucleotide primers that were complementary to the same regions of both viruses. The sequences flanked by the primers were unique for each virus and could be differentiated by hybridization with 40-base, 32P-labeled oligonucleotide probes or by cleavage with BamHI. The DNA fragments resulting from amplification of BKV and JCV were 176 and 173 nucleotide pairs, respectively. The sensitivities of PCR for amplification of cloned BKV and JCV DNAs were 10 and 100 copies, respectively. Hybridization with the oligonucleotide probes was specific for each virus. A total of 57 urine samples from three groups of subjects were processed by DNA extraction or boiling and were tested by PCR. Urine samples collected from immunosuppressed patients (n = 11) and previously documented to be positive for BKV, JCV, or both were positive by PCR. Ten percent of urine samples from healthy adults (n = 30) that were previously negative for BKV and JCV were positive for one or both viruses by PCR. Urine samples (n = 16) from four seronegative bone marrow transplant recipients were uniformly negative for BKV. JCV was detected in deparaffinized brain tissue from a patient with progressive multifocal leukoencephalopathy. Specific diagnosis of virus in clinical specimens could be made within 1 day of receipt of the specimens. The PCR method is attractive for use in diagnosing polyomavirus infections because of its sensitivity, specificity, and rapid turnaround time.
Evaluation of oligonucleotide probes for identification of shiga-like-toxin-producing Escherichia coli
Authors: Karch H , Meyer T
Published: January 2020

Four synthetic oligonucleotide probes representing different regions of the Shiga-like toxin I (SLT-I) structural genes and one oligonucleotide derived from the SLT-II gene of Escherichia coli serotype O157:H7 strain 933 were examined for the identification of E. coli strains that produce cytotoxins for Vero or HeLa cells. E. coli strains that synthesize SLT-I alone or O157:H7 isolates that coexpress SLT-I and SLT-II hybridized with all four probes that were complementary to the SLT-I genes, suggesting that they have toxin genes with great homology in all the regions examined. In colony hybridization tests, these oligonucleotide probes did not react with E. coli strains that were nontoxigenic for Vero cells or that produced cytotoxins belonging to the SLT-II family. The probe derived from the slt-IIA gene distinguished E. coli strains that produced SLT-II alone from SLT-I-producing strains and hybridized to all E. coli O157:H7 strains that produced both SLT-I and SLT-II. Using two of these oligonucleotide probes that were complementary to slt-IA or slt-IIA sequences, we identified 50 of 52 cytotoxin-producing strains, whereas none of 416 nontoxigenic E. coli strains was reactive. The colony blot hybridization with the oligonucleotide probes described here can serve as a specific and sensitive test with potential diagnostic value.
Cluster of Malassezia furfur pulmonary infections in infants in a neonatal intensive-care unit
Authors: Richet H M , McNeil M M , Edwards M C , Jarvis W R
Published: January 2020

Between 23 and 27 July 1987, three infants at one hospital developed severe bronchopneumonia associated with respiratory failure, thrombocytopenia, and leukocytosis. Two infants died; at postmortem examination, Malassezia furfur was identified in their lung tissues. M. furfur was isolated from cultures of blood, urine, and stool samples from the infant who survived. All documented M. furfur infections occurred in infants with a birth weight of less than 1,000 g; the attack rate was 42.9% (three of seven infants). A case-control study comparing the three cases and nine infants randomly selected from infants in the neonatal intensive care unit during the outbreak showed the following variables to be significantly associated with case-infants: younger gestational age (less than 26 weeks), hyaline membrane disease, duration of ventilation, duration of antimicrobial therapy, and the presence of a Broviac catheter. In a second case-control study, in which case-infants were compared with birth weight-matched controls, only the duration of antimicrobial therapy was significantly associated with case-infants. A point prevalence culture survey showed that 2 of 10 infants and 2 of 11 personnel were colonized with M. furfur. This cluster suggests that M. furfur can be transmitted from an infected or colonized infant to other infants. Infection control practices should be aimed at (i) identifying high-risk infants and (ii) reemphasizing the importance of hand washing.
Clinical and microbiological evaluation of four culture methods for the diagnosis of peritonitis in patients on continuous ambulatory peritoneal dialysis
Authors: Doyle P W , Crichton E P , Mathias R G , Werb R
Published: January 2020

A prospective study was performed to evaluate four culture methods for the diagnosis of bacterial peritonitis in patients on continuous ambulatory peritoneal dialysis. Peritonitis was present in 44 of 85 patient admissions (52%). The overall sensitivity of the culture methods in detecting peritonitis was 66%. The sensitivities of the individual methods were as follows: bag culture method, 61%; blood culture broth method, 51%; filter method, 54%; and plate method, 39%. Our broad definition of peritonitis resulted in lower sensitivities. A combination of the bag and blood culture broth methods detected all positive cultures.
Efficacy of duplicate genital specimens and repeated testing for confirming positive results for chlamydiazyme detection of Chlamydia trachomatis antigen
Authors: Kellogg J A , Seiple J W , Levisky J S
Published: January 2020

In an attempt to increase Chlamydiazyme (Abbott Laboratories) detection of Chlamydia trachomatis antigen and to establish the reproducibility of positive results, we carried out an investigation into the usefulness of testing duplicate specimens, of more aggressive endocervical specimen collection by using cytobrushes instead of swabs, and of the repeated testing of both specimens from patients with one or two positive results. Duplicate endocervical (female) and urethral (male) specimens, including one swab and one cytobrush specimen from 1,331 nonpregnant women, were collected from symptomatic and asymptomatic patients. Specimens were transported and tested for C. trachomatis antigen as specified by the manufacturer. Tests on all specimens from patients with positive results were repeated. Antigen was initially detected in one or both specimens from 210 (10.7%) of 1,968 patients, and repetition of the tests confirmed its presence in 198 (10.1%) of the patients, including all 183 patients in whom it was initially detected in both specimens. Initial results from at least 8 of the 12 patients with irreproducible antigen detection were most probably falsely positive. Results from 21 (10.6%) of the 198 patients for whom antigen detection was confirmed were repeatedly positive on only one specimen (9 [4.5%] on the second of the two specimens collected). Of 115 women from whom one swab and one cytobrush sample were taken and who had repeatedly positive results, antigen was detected in 7 (6.1%) only on the swab sample and in 4 (3.5%) only on the cytobrush sample. Use of the cytobrush does not appear justified with the Chlamydiazyme assay, and collection of duplicate specimens provided only a modest increase in detection of C. trachomatis. However, repeated testing of specimens when results from only one of two specimens are positive appears to be of clinical value.
Antilipopolysaccharide antibodies and differential diagnosis of chronic Pseudomonas aeruginosa lung infection in cystic fibrosis
Authors: Fomsgaard A , Dinesen B , Shand G H , Pressler T , Høiby N
Published: January 2020

Chronic lung infection in cystic fibrosis is characteristically associated with polyagglutinable, serum-sensitive, mucoid strains of Pseudomonas aeruginosa. Enzyme-linked immunosorbent assay (ELISA) methods for standard-free quantitation of immunoglobulin G (IgG) and IgM antibodies to P. aeruginosa lipopolysaccharides (LPSs) have been developed. We now report the development of assays for quantitation of monomer and dimer total IgA and IgA anti-LPS antibodies. Use of these methods in diagnosis of early chronic P. aeruginosa lung infection was assessed. IgG and IgA anti-LPS levels increased significantly at the onset of chronic infection and continued to increase to very high levels in the later stages of infection. IgM anti-LPS levels also rose at the onset of chronic infection but did not increase further. The function of true- and false-positive rates was illustrated by using various concentrations of IgG, IgA, and IgM anti-LPS for discrimination of patients. Values that gave optimum separations were used for statistical evaluation of the diagnostic sensitivities and specificities of anti-LPS antibody concentrations. The results obtained in these assays were compared with a diagnosis, based on the number of precipitins in crossed immunoelectrophoresis, of serum samples from cystic fibrosis patients. In 64 paired serum samples taken before and immediately after the onset of chronic infection, as defined by crossed immunoelectrophoresis precipitins, the predictive values of a positive ELISA were 86% for IgG and 89% for IgA. The predictive values for a negative ELISA were 98% for IgG and 97% for IgA. Results of the IgM anti-LPS ELISA had a lower predictive value. Immunoblotting and absorption studies showed that IgG anti-LPS antibodies were directed specifically against LPS of P. aeuruginosa. ELISAs were developed to determine the specific IgG sublclasses involved. The increase in IgG anti-LPS involved all four subclasses. Highest anti-LPS titers were seen with IgG1 and IgG4, but the largest relative increases were seen with IgG2 and IgG3.
Agreement study between two laboratories of immunofluorescence as a confirmatory test for human immunodeficiency virus type 1 antibody screening
Authors: Mahony J , Rosenthal K , Chernesky M , Castriciano S , Scheid E , Blajchman M , Harnish D
Published: January 2020

A total of 114 serum specimens from 76 blood donors, 21 patients with acquired immune deficiency syndrome or acquired immune deficiency syndrome-related complex, 7 multiply transfused patients, 3 hemophiliacs, and 7 others were tested for anti-human immunodeficiency virus type 1 (HIV-1) antibody by enzyme immunoassay (EIA) and Western blot (WB) and then blindly tested by immunofluorescence (IF), independently, in two separate laboratories. The IF technique used acetone-fixed HIV-1-infected E cells and uninfected HUT-78 cells mixed at a 1:3 ratio in one spot on a glass slide and uninfected HUT-78 cells (to assess nonspecific fluorescence) alone in a second spot. Of 114 serum specimens, 85 were repeat EIA positive, and 21 of these were WB positive. A total of 129 of 134 of the IF results (included were 20 duplicates) were identical between laboratories, for a Kappa agreement statistic of 0.93. All five IF results discordant between laboratories were EIA repeat positive and WB negative. Included in the study were eight WB-indeterminate sera, of which five blood donor serum specimens and one hemophiliac serum specimen were IF negative and two acquired immune deficiency syndrome serum specimens were IF positive. As a confirmatory test for HIV-1 antibodies, IF provided a faster alternative or supplementary test for confirming EIA results.
Comparison of monoclonal antibody time-resolved fluoroimmunoassay with monoclonal antibody capture-biotinylated detector enzyme immunoassay for respiratory syncytial virus and parainfluenza virus antigen detection
Authors: Hierholzer J C , Bingham P G , Coombs R A , Johansson K H , Anderson L J , Halonen P E
Published: January 2020

An all-monoclonal antibody, time-resolved fluoroimmunoassay was compared with several enzyme immunoassays for the detection of respiratory syncytial virus and parainfluenza virus type 1, 2, and 3 antigens in clinical specimens. The most sensitive enzyme immunoassay for parainfluenza virus type 1 was an all-monoclonal antibody assay with biotin-labeled detector antibody and streptavidin-peroxidase conjugate, but for respiratory syncytial virus and parainfluenza virus types 2 and 3 the most sensitive assay was a polyclonal antibody assay with horse capture antibodies and bovine or rabbit detector antibodies with anti-species peroxidase. All tests were evaluated with nasopharyngeal aspirate specimens from respiratory illnesses and with cell culture harvests of multiple strains of each virus isolated over many years. The time-resolved fluoroimmunoassay detected respiratory syncytial virus antigen in 92% of the specimens positive by culture, which was a decidedly higher sensitivity than either the monoclonal or polyclonal antibody enzyme immunoassay format (62 and 76%, respectively). For the parainfluenza viruses the time-resolved fluoroimmunoassay detected type-specific antigen in 94 to 100% of culture-positive specimens and again was more sensitive than the all-monoclonal antibody enzyme immunoassays (75 to 89%) or all-polyclonal antibody enzyme immunoassays (66 to 95%). Combined with results from a previously reported adenovirus time-resolved fluoroimmunoassay, these tests identified respiratory antigens in large numbers of clinical specimens.
Indirect immunoglobulin G (IgG) and IgM enzyme-linked immunosorbent assays (ELISAs) and IgM capture ELISA for detection of antibodies to lipopolysaccharide in adult typhoid fever patients in Pakistan
Authors: Sippel J , Bukhtiari N , Awan M B , Krieg R , Duncan J F , Karamat K A , Malik I A , Igbal L M , Legters L
Published: January 2020

Sera from 339 adult febrile patients in Pakistan were tested for antibodies to Salmonella typhi lipopolysaccharide by indirect immunoglobulin G (IgG) and IgM enzyme-linked immunosorbent assay (ELISA) and IgM capture ELISA. A total of 55 patients had S. typhi cultured from their blood, 20 had S. typhi cultured from their stool, 24 were blood or stool culture positive for S. paratyphi A, 41 were culture negative but clinically diagnosed as having enteric fever, 41 had gastrointestinal or urinary tract infections, 41 were clinically diagnosed as having malaria, 20 were smear-positive patients with malaria, 58 had respiratory infections, and the remaining 39 individuals were placed in a miscellaneous group who did not have Salmonella infection. The sensitivities of the indirect IgG ELISA, indirect IgM ELISA, and IgM capture ELISA determined with specimens obtained from the blood culture-positive patients with typhoid fever (positive controls) were 80, 64, and 62%, respectively. The specificities of the assays determined with sera from the patients with respiratory infections (negative controls) were 95, 95, and 97%, respectively. The percentage of smear-positive patients with malaria who were positive by these assays was lower than that in the negative control group. The percentages of individuals in the other patient categories who were positive by these tests were between those obtained with the positive and negative controls. Of the positive controls, 26 were positive by both IgM assays, 9 were IgM positive only by indirect ELISA, and 8 were IgM positive only by IgM capture ELISA. A total of 70% of the positive control patients who were tested for O agglutinins by the Widal tube agglutination assay were positive; however, 29% of the negative control patients were also positive. The indirect IgG ELISA was the single most effective test for the serodiagnosis of typhoid fever in this population.
Oral Candida albicans isolates from nonhospitalized normal carriers, immunocompetent hospitalized patients, and immunocompromised patients with or without acquired immunodeficiency syndrome
Authors: Brawner D L , Cutler J E
Published: January 2020

A total of 128 human oral isolates of Candida albicans were collected from asymptomatic healthy carriers (64 isolates); asymptomatic, nonimmunosuppressed, hospitalized patients (25 isolates); immunosuppressed transplant patients (19 isolates); and human immunodeficiency virus-infected patients with symptoms of acquired immunodeficiency syndrome and oral candidiasis (20 isolates). Isolates were serotyped as A or B and tested for reactivity with an agglutinating immunoglobulin M monoclonal antibody (H9). Immunocompetent individuals colonized by oral C. albicans were almost equally likely to carry serotype A as serotype B cells, while immunocompromised individuals were at least twice as likely to be infected by serotype B than serotype A strains. The reactivity of isolates with H9 antibody followed a similar but more distinctive pattern. Approximately half of the strains from immunocompetent individuals reacted strongly with H9, and the remainder reacted weakly. However, up to 75% of the isolates from immunocompromised patients reacted weakly with H9, while the remainder reacted strongly. A correlation between H9 reactivity and the serotypes of these isolates existed (P = 0.16). The correlation between H9 reactivity and immune status was even stronger (P = 0.025). The monoclonal antibody activities described above were determined by agglutination tests during defined phases of C. albicans growth. Expression of antigen at various times during growth of several isolates was confirmed at the cellular level by analysis using fluorescence-activated cell sorting. Despite the correlation between serotype A and H9 reactivity, H9 antigen was not identical to the serotype A antigen because four serotype A strains reacted only weakly with H9 antibody, and one strain reacted strongly with H9 but was serotype B. These data indicate that oral strains of C. albicans from immunocompetent individuals differ as a group from C. albicans isolated from those who are immunosuppressed.
Clinical impact of rapid identification and susceptibility testing of bacterial blood culture isolates
Authors: Trenholme G M , Kaplan R L , Karakusis P H , Stine T , Fuhrer J , Landau W , Levin S
Published: January 2020

Two hundred twenty-six patients with bacteremia were prospectively enrolled in a randomized trial that was performed to determine the clinical impact of the receipt of in vitro microbiological data by the physician soon after organism detection as opposed to having the physicians wait until similar data were available by routine methods. Identification and antibiotic susceptibility patterns of 110 isolates were determined by direct inoculation of the Vitek AutoMicrobic system (Vitek Systems, Inc., Hazelwood, Mo.) with a sample from a positive blood culture vial. One hundred sixteen isolates were processed by routine methods. Microbiological results were available within an average of 8.8 h by the direct method versus an average of 48 h by the routine method. In both groups an infectious disease fellow used the information to make therapeutic recommendations to the responsible physician. When compared with that provided by the routine method, the information provided by the direct method was significantly more likely to result in an initiation of antibiotic therapy, a change to more effective therapy, or a change to less expensive therapy. Recommendations were significantly more likely to be followed in patients whose isolates were processed by the direct method versus the routine method. A projected savings of $158 per patient was estimated for the patients who were changed to less expensive therapy or in whom antibiotics were discontinued because results were available sooner. These cost savings, coupled with changes in therapy made for reasons of efficacy, support the usefulness of the earlier reporting of the identity and antibiotic susceptibility patterns of bacterial blood culture isolates.
Staphylococcus aureus strains which are not identified by rapid agglutination methods are of capsular serotype 5
Authors: Fournier J M , Boutonnier A , Bouvet A
Published: January 2020

A total of 183 recent Staphylococcus aureus clinical isolates were tested with three commercially available rapid agglutination methods. The capsular polysaccharide type and resistance to oxacillin of these isolates were also determined. Seven isolates were not identified correctly by agglutination methods. All isolates not identified by the rapid methods were of capsular serotype 5, and of these isolates, six were resistant to oxacillin. The results suggest that these agglutination kits can be improved by the use of antibodies reactive with S. aureus capsular polysaccharide.
Virulence of Campylobacter jejuni for chicken embryos
Authors: Mahajan S , Rodgers F G
Published: January 2020

The pathogenicity of Campylobacter jejuni was examined in chicken embryos. In this system, mortality data and histopathological findings induced by organisms and by bacterium-free filtered broth were identical. The absence in chicken embryo tissues both of organisms and of an inflammatory infiltrate suggests a toxin etiology.
Morphologic study of Staphylococcus aureus L-form, reverting, and intermediate colonies in situ
Authors: Owens W E , Nickerson S C
Published: January 2020

Staphylococcus aureus strains of bovine origin were induced to L-form by exposure to 100 U of penicillin in brain heart infusion broth supplemented with 5% NaCl, 5% sucrose, and 10% horse serum. L-forms were cultured on similarly supplemented brain heart infusion agar containing no antibiotic. Light and electron microscopic examination of plastic-embedded L-form colonies revealed a variety of morphologic types. The primary site of growth appeared to be the core area below the agar surface, consisting mainly of pleomorphic budding forms. At the surface, these forms gave rise to large spherules with a gradation from smaller to larger spherules toward the periphery of the colony. Some colonies progressed to reverting forms with the growth of bacterial cells containing cell wall. In addition to L-forms, intermediate colony forms were observed that lacked typical L-form morphology and progressed rapidly to the parent cell form on subculture to bovine blood agar. Description of these forms will be used in the search for similar morphologic types in vivo during antibiotic treatment of chronic S. aureus bovine mastitis.
Comparison of HeLa 229 and McCoy cell cultures for detection of Chlamydia trachomatis in clinical specimens
Authors: Thewessen E A , Freundt I , van Rijsoort-Vos J H , Stolz E , Michel M F , Wagenvoort J H
Published: January 2020

Consecutive clinical specimens of Chlamydia trachomatis (1,048) were inoculated in parallel on DEAE-dextran- and cycloheximide-treated HeLa 229 cells and cycloheximide-treated McCoy cells. HeLa 229 cell culture detected 113 positive specimens, and McCoy cell culture detected 103 positive specimens. This difference is not significant. However, HeLa 229 cell culture yielded significantly more inclusions than McCoy cell culture in the 95 specimens positive in both cell types (P = 0.042). For routine diagnostic purposes, a choice for one of the cell types may be determined by local preferences.