bluuebunny/crossref_metadata_2025 huggingface dataset:
Purification and characterization of an 80-kilodalton Trypanosoma cruzi urinary antigen
Authors: Corral R S , Orn A , Freilij H L , Bergman T , Grinstein S
Published: January 2020

A Trypanosoma cruzi antigen eliminated in the urine of experimentally infected dogs was detected by enzyme-linked immunosorbent assay between 9 and 28 days after infection. The parasite urinary antigen (UAg) was purified by affinity chromatography with polyclonal antibodies to T. cruzi. The eluate of the antibody column was subjected to high-performance liquid chromatography and showed a single peak of A280. This antigen was the only parasite component found in the urine of infected dogs during the course of acute T. cruzi infection. Antigen characterization was performed by two-dimensional gel electrophoresis, lectin affinity chromatography, proteolytic digestion, and Western blotting (immunoblotting). The isolated UAg exhibited a relative molecular size of 80 kilodaltons (kDa), an isoelectric point of 6.2 to 6.8, binding to concanavalin A, and sensitivity to trypsin. The parasite antigen was electroeluted from polyacrylamide gels and subjected to acid hydrolysis and amino acid analysis by reverse-phase high-performance liquid chromatography. The 80-kDa glycoprotein was recognized by serum antibodies from a wide variety of T. cruzi-infected hosts. The UAg proved to be a highly antigenic component present in different strains of T. cruzi. This 80-kDa polypeptide resembles one of the parasite antigens previously found in the urine of patients with acute Chagas' disease.
An unusual strain of Legionella micdadei
Authors: Pastoris M C , Collins M T , Baldassarri L , Maggio A
Published: January 2020

A microorganism antigenically identified as Legionella micdadei but showing a cellular fatty acid profile distinct from that described previously for this species and more similar to that of L. bozemanii has been studied by phenotypic characterization, crossed immunoelectrophoresis, gas-liquid chromatography, and transmission electron microscopy. Although the phenotypic characters, the crossed immunoelectrophoresis, and the ultrastructural features were similar to those of L. micdadei, the fatty acid composition differed significantly from this species; moreover it differed also from L. bozemanii, even though it was quantitatively more similar.
Evaluation of NCS rapid plasma reagin card test, a new screening kit for syphilis
Authors: Fischer G S , Colavita M T , Sweimler W I , Kleger B
Published: January 2020

Agreements in qualitative and quantitative test results between the recently marketed NCS rapid plasma reagin card test (NCS Diagnostics Corp.) and the Macro-Vue rapid plasma reagin 18-mm circle card test (Hynson, Westcott and Dunning) were 99.2 and 99.1%, respectively, indicating the NCS RPR card test to be an acceptable alternative procedure for the serodiagnosis of syphilis.
Cloned Schistosoma mansoni proteinase (hemoglobinase) as a putative serodiagnostic reagent
Authors: Chappell C L , Hackel J , Davis A H
Published: January 2020

Expressed cDNA encoding a proteolytic enzyme from Schistosoma mansoni has been cloned recently. Circulating antibodies reacting with the recombinant protein have been detected in the blood of mice and humans infected with S. mansoni, S. japonicum, or S. haematobium. S. mansoni and S. haematobium infection can be distinguished by antibody titer.
Comparison of the virologic and immunologic responses of volunteers to live avian-human influenza A H3N2 reassortant virus vaccines derived from two different avian influenza virus donors
Authors: Clements M L , Sears S D , Christina K , Murphy B R , Snyder M H
Published: January 2020

We compared the abilities of the six internal RNA segments of two avian influenza viruses, A/Mallard/Alberta/88/76 (H3N8) and A/Mallard/NY/6750/78 (H2N2), to confer attenuation on wild-type human influenza A/Bethesda/1/85 (H3N2) virus in seronegative adult volunteers. Live avian-human influenza A reassortant virus vaccines derived from either avian virus parent were comparable in the following properties: safety, infectivity, immunogenicity, and genetic stability. Since the avian influenza A/Mallard/Alberta/76 virus offered no clear advantage as a donor virus, we will conduct our future evaluations on live influenza A virus reassortants derived from the more extensively characterized avian influenza A/Mallard/NY/78 virus.
Ofloxacin susceptibility testing quality control parameters for microdilution and disk diffusion, and confirmation of disk diffusion interpretive criteria
Authors: Fuchs P C , Jones R N , Barry A L , Gavan T L
Published: January 2020

The susceptibilities of 221 clinical isolates to ofloxacin were tested simultaneously by broth microdilution and disk diffusion methods with commercially prepared 5-micrograms ofloxacin disks. The acceptability of the following previously proposed zone diameter breakpoints was confirmed: greater than or equal to 16 mm, susceptible; 13 to 15 mm, intermediate; less than or equal to 12 mm, resistant. On the basis of a multilaboratory collaborative study, the following are proposed as acceptable ofloxacin MIC ranges for quality control organisms: Escherichia coli ATCC 25922, 0.03 to 0.06 micrograms/ml; Staphylococcus aureus ATCC 29213, 0.12 to 0.5 micrograms/ml; Pseudomonas aeruginosa ATCC 27853 and Enterococcus faecalis ATCC 29212, 1.0 to 4.0 micrograms/ml. Ofloxacin quality control zone diameter ranges for the disk diffusion test are tentatively proposed, but variations in the performance of different lots of Mueller-Hinton agar may prove to be a serious problem for users.
Detection of human rotavirus by using an alkaline phosphatase-conjugated synthetic DNA probe in comparison with enzyme-linked immunoassay and polyacrylamide gel analysis
Authors: Olive D M , Sethi S K
Published: January 2020

An alkaline phosphatase-conjugated synthetic oligodeoxyribonucleotide probe was compared with polyacrylamide gel electrophoresis (PAGE) detection of rotavirus RNA as well as an enzyme-linked immunosorbent assay (ELISA) for the detection of rotavirus in stools from young children with gastroenteritis. The synthetic probe did not cross-react with bacterial causative agents of diarrheal disease. Extraction of viral RNA from stool samples with a phenol-chloroform mixture was suitable for most samples. In some cases fecal pigments interfered with the reaction of the probe with viral RNA. The use of ion-exchange chromatography to further purify viral RNA removed contaminating pigments and increased the sensitivity of the probe assay. Of 260 stool specimens, 77 (30%) were positive for rotavirus when tested by PAGE analysis of rotavirus RNA. The synthetic probe identified 71 rotavirus specimens when RNA obtained by phenol-chloroform extraction followed by chromatographic purification was used (sensitivity, 91.0%; specificity, 96.7%). The ELISA results also agreed well with the electrophoretic analysis (sensitivity, 98.7%; specificity 94%) and the probe assay (sensitivity, 90%; specificity, 100%). Discordant results between the ELISA and the probe assay were examined further by electron microscopy and PAGE analysis of viral RNA. The positive and negative predictive values of the probe assay in comparison with PAGE were 92.2 and 96.1%, respectively. Rotaviruses showing both long and short RNA electrophoretic patterns were detected by the probe. The probe assay coupled with chromatographic purification of rotavirus RNA is an effective method for detecting rotavirus and compares favorably with PAGE analysis and ELISA.
Staphylococcus intermedius in canine gingiva and canine-inflicted human wound infections: laboratory characterization of a newly recognized zoonotic pathogen
Authors: Talan D A , Staatz D , Staatz A , Goldstein E J , Singer K , Overturf G D
Published: January 2020

Staphylococcal gingival flora was characterized in cultures from 135 dogs. Staphylococcus intermedius was isolated in 39% of the cultures, S. aureus was isolated in 10%, and both were isolated in 2.0%. S. aureus was isolated more often from dogs of working breeds with weights of greater than 40 lb (ca. 18 kg) and with outdoor habitats than was S. intermedius, which was associated with dogs of nonworking breeds with weights of less than 40 lb and indoor habitats. S. intermedius was distinguished from S. aureus by the following characteristics: coagulation of rabbit plasma at 4 h (26 versus 100%, respectively), hemolysis of sheep blood at 24 h (30 versus 79%, respectively), and mannitol fermentation at 24 h (4 versus 93%, respectively). A clear separation of the two species was apparent only with the acetoin (modified Voges-Proskauer) reaction (100% of the S. aureus isolates versus 0% of the S. intermedius isolates) and beta-galactosidase activity on the API Staph-Ident strip (0% of the S. aureus isolates and 100% of the S. intermedius isolates). Susceptibilities of S. intermedius and S. aureus were 72 and 7%, respectively, to penicillin G, and 100% of both species to oxacillin. Fourteen previously collected strains of coagulase-positive staphylococci from infected canine-inflicted human wounds were reanalyzed; 3 of 14 (21%) isolates were S. intermedius. We conclude that S. intermedius is a common canine gingival flora and is responsible for some canine-inflicted human wound infections, thus representing a newly recognized zoonotic pathogen.
Isolation and characterization of Klebsiella pneumoniae unencapsulated mutants
Authors: Benedí V J , Ciurana B , Tomás J M
Published: January 2020

Klebsiella pneumoniae mutants were obtained after UV irradiation and negative selection with anticapsular serum. Unencapsulation, rather than expression of a structurally altered capsule, was found in the mutants. The mutant strains showed no alterations in their outer membrane proteins and lipopolysaccharide, and a great similarity with the wild type in the properties tested (serum resistance, antimicrobial sensitivity, and lipopolysaccharide-specific bacteriophage sensitivity), with the exception of a higher cell surface hydrophobicity and resistance to bacteriophage FC3-9.
Isolation of human immunodeficiency virus from peripheral blood lymphocytes stored in various transport media and frozen at -60 degrees C
Authors: Gallo D , Kimpton J S , Johnson P J
Published: January 2020

The peripheral blood lymphocytes from 48 heparinized blood specimens from human immunodeficiency virus (HIV) antibody-positive individuals were divided into two aliquots. One aliquot was reconstituted in one of the following five media. Medium 1 consisted of tryptose broth with 0.5% gelatin; medium 2 consisted of RPMI 1640 containing 10% fetal bovine serum (FBS); medium 3 consisted of RPMI 1640 containing 20% FBS, Polybrene, interleukin 2, and anti-alpha interferon; medium 4 consisted of medium 2 plus 10% dimethyl sulfoxide (DMSO); and medium 5 consisted of medium 3 plus 10% DMSO. Lymphocytes were stored in these five media at -60 degrees C. The other aliquot of cells was stored at -190 degrees C in RPMI 1640 containing 50% FBS and 10% DMSO. After 1 week, both aliquots were cocultivated with phytohemagglutinin-stimulated uninfected peripheral blood lymphocytes, and presence of HIV was detected by the reverse transcriptase test. Storage in medium 1, 2, or 3 did not result in satisfactory isolation rates, but storage at -60 degrees C in medium 4 or 5 gave equal or better isolation rates than did storage at -190 degrees C. Inactivation of HIV by freezing of the cells without DMSO correlated with high antibody titers to core and polymerase proteins as measured by Western (immuno-) blotting.
Pneumococcosuria in children
Authors: Miller M A , Kaplan B S , Sorger S , Knowles K F
Published: January 2020

Streptococcus pneumoniae was present, in pure or mixed culture, in 43 (0.08%) of 53,499 urine cultures submitted from a pediatric population over a 4-year period. Data were analyzed from 28 children, from whom 78% of these positive cultures originated. Ninety-six percent of the children were female, and the median age was 3 years (range, 0.4 to 17 years). Only five children had S. pneumoniae as the sole organism cultured from the urine, and all five had only a single urine culture. Urine from the other 23 children contained other organisms as well. Small numbers of pneumococci were found in most urines: 74% contained less than 10(4) CFU/ml and 93% contained less than 10(5) CFU/ml. There was no association between genitourinary symptoms and pneumococcosuria, and complete resolution of symptoms occurred in both treated and untreated children. Pneumococcosuria could not be explained by pneumococcal bacteremia. We conclude that pediatric pneumococcosuria is not associated with urinary tract infections in children, or with pneumococcal bacteremia or invasive disease elsewhere. Pneumococcosuria probably reflects contamination of urine specimens with S. pneumoniae from perineal colonization.
Virulence factors and biochemical characteristics of serotypes of Escherichia coli serogroup O29
Authors: Guth B E , Silva R M , Toledo M R , Lima T M , Trabulsi L R
Published: January 2020

Escherichia coli strains belonging to serogroup O29 were studied. Invasiveness was the most common virulence factor described in this serogroup, but a few papers also reported production of heat-stable (ST) enterotoxin. In the present study invasive ability was found in O29:H- strains, whereas production of ST-I enterotoxin was observed only in serotype O29:H21 strains, showing that virulence was a characteristic of specific serotypes or bioserotypes within the O29 serogroup. Different serotypes were found among strains that were neither invasive nor toxigenic. Invasive strains were biochemically less active than the toxigenic ones and presented the invasiveness plasmid (pINV) of about 120 to 140 megadaltons, whereas hybridization tests showed that ST-I production was related to a plasmid of about 90 megadaltons. A diffuse adherence pattern to HeLa cells was observed in all ST-I isolates, but the role of this adherence in the pathogenicity of these strains was not determined. Thus, a unique biochemical pattern and plasmid profile may be useful characteristics to distinguish between pathogenic (toxigenic or invasive) and nonpathogenic O29 strains.
Pasteurella caballi, a new species from equine clinical specimens
Authors: Schlater L K , Brenner D J , Steigerwalt A G , Moss C W , Lambert M A , Packer R A
Published: January 2020

The name Pasteurella caballi is proposed for a group of organisms represented by 29 strains isolated from respiratory and other infections in horses. P. caballi strains are gram-negative, oxidase-positive, nonmotile, fermentative rods with the key characteristics of the genus Pasteurella. These strains differed from other Pasteurella species in that all were aerogenic and catalase negative, and some strains produced acid from myo-inositol and L-rhamnose. The levels of DNA relatedness of 28 P. caballi strains with labeled DNA from the proposed type strain averaged 91 and 85% (hydroxyapatite method at 55 and 70 degrees C). P. caballi was 13 to 53% related to strains representing 22 other species of the family Pasteurellaceae. The guanine-plus-cytosine content of the DNA of four strains was 41 to 42 mol%. The type strain is 83851 (=ATCC 49197).
Characterization of coagulase-negative staphylococci by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analyses
Authors: Thomson-Carter F M , Pennington T H
Published: January 2020

Coagulase-negative staphylococci are important nosocomial pathogens. At present, no wholly satisfactory typing scheme exists for these organisms. Therefore, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting were assessed as characterization methods. A total of 100 type strains and nontyped isolates representing nine species of coagulase-negative staphylococci were analyzed. Each species had a reproducible, characteristic whole-cell banding pattern when analyzed by either method. These species-specific profiles were obtained for all isolates despite disparate geographical origins and clinical isolation sites. Intraspecies similarities, calculated by using the Dice coefficient, were significantly higher than interspecies similarities. Although some species were more heterogeneous than others, the allocation of isolates to any particular species was reinforced by the high degree of interspecies dissimilarity. Application of SDS-PAGE also distinguished discrete subspecies groups. These groups possessed the characteristic profile of their species but were distinguished by a group of variable polypeptides. Species-specific banding patterns were also obtained by immunoblotting of whole-cell polypeptides. Differences between immunoblot and SDS-PAGE profiles could be attributed to variations of antigenicity of particular polypeptides. However, both SDS-PAGE and immunoblotting provided reproducible and sensitive methods for characterization of coagulase-negative staphylococci. Standardization of these techniques could provide the basis for a primary typing scheme.
Establishment of titration system for human herpesvirus 6 and evaluation of neutralizing antibody response to the virus
Authors: Asada H , Yalcin S , Balachandra K , Higashi K , Yamanishi K
Published: January 2020

The susceptibilities of seven T-cell lines to human herpesvirus 6 (HHV-6) infection were examined. MT-4 cells were the most susceptible of these lines to infection with this virus. Therefore, chemically adhered MT-4 cell monolayers were used for infectious HHV-6 assay by indirect immunofluorescent-antibody (IFA) staining. When cell monolayers were fixed 30 to 45 h postinfection, the foci stained with IFA were easy to count and a linear relationship was observed between the number of foci and the virus concentration. MT-4 cell monolayers were also used for a focus reduction neutralizing-antibody test. In this test, sera from patients in the convalescent stage of exanthem subitum all showed significant neutralizing activity (1:80 to 1:320), whereas sera from patients in the acute stage of disease showed no detectable neutralizing activity. The titers of neutralizing antibody correlated well with the levels of anti-HHV-6 antibodies detected by IFA.
Investigation of apparent false-positive urine latex particle agglutination tests for the detection of group B streptococcus antigen
Authors: Harris M C , Deuber C , Polin R A , Nachamkin I
Published: January 2020

In our nursery, we identified neonates with positive urine latex particle agglutination (LPA) tests for group B streptococcus (GBS) antigen who did not have corroborating cultural evidence of infection. To investigate the mechanisms underlying these apparent false-positive reactions, we examined the urine LPA test in an unselected population of neonates suspected of sepsis. Urine specimens for LPA testing and culture and simultaneous perirectal cultures were obtained from 134 neonates with suspected sepsis. Six infants had positive blood cultures for GBS; four of the six were positive by LPA testing. An additional 20 infants had positive LPA tests but negative blood cultures; of these, 13 had mothers who received antibiotic treatment prior to delivery. Two infants with positive LPA results and negative blood cultures had bacteria isolated from urine cultures obtained in a nonsterile fashion (GBS, Escherichia coli). GBS was not isolated from perirectal swabs of infants with positive LPA tests and negative blood cultures. In conclusion, (i) a high proportion of neonates evaluated for sepsis gave positive LPA tests and negative blood cultures, (ii) local contamination of the perirectal skin or urinary tract with GBS was an unlikely source of false-positive LPA reactions, and (iii) maternal antibiotic pretreatment during labor may represent an important cause of apparent false-positive LPA reactions.
Two-dimensional gas chromatography with electron capture detection for the sensitive determination of specific mycobacterial lipid constituents
Authors: Larsson L , Jimenez J , Sonesson A , Portaels F
Published: January 2020

A method was developed for determining two characteristic mycobacterial lipid constituents, tuberculostearic acid (as its pentafluorobenzyl ester) and 2-eicosanol (as its pentafluorobenzoyl ester), by using gas chromatography with electron capture detection. A microprocessor-controlled column-switching system (two-dimensional gas chromatography) facilitated sample preparation and increased specificity. The usefulness of the technique was illustrated by its ability to reveal picogram amounts of tuberculostearate in a suspension of Mycobacterium leprae isolated from a naturally infected armadillo. Two-dimensional gas chromatography with electron capture detection may in some instances provide a convenient alternative to gas chromatography-mass spectrometry for use in demonstrating the presence of mycobacteria in a complex environment.
Serotype, antimicrobial resistance, and adherence properties of Escherichia coli strains associated with outbreaks of diarrheal illness in children in the United States
Authors: Moyenuddin M , Wachsmuth I K , Moseley S L , Bopp C A , Blake P A
Published: January 2020

Since most recorded outbreaks of diarrhea in U.S. infants attributed to Escherichia coli occurred before currently available pathogenicity assays existed, we examined the characteristics of nonenterotoxigenic E. coli strains isolated from 50 outbreaks of diarrheal disease in U.S. infants between 1934 and 1987. We assayed the strains for enteropathogenic E. coli (EPEC) serotype, localized adherence (LA) and diffuse adherence to tissue cultures, the presence of EPEC adherence factor genes, Shiga-like (Vero) toxin production, and antimicrobial resistance. EPEC serotypes were identified in 28 outbreaks (56%). LA to HeLa cells was found in 23 outbreak strains and correlated 100% with the EPEC adherence factor probe. LA was observed in 21 of 28 EPEC and 2 of 22 non-EPEC strains; however, 5 of 23 strains that were LA positive for HeLa cells did not adhere to HEp-2 or HL cells. One strain was diffuse adherence positive, and none was Shiga-like toxin positive. Multiple resistance was common in EPEC (64%), LA-positive (74%), and LA-positive EPEC (76%) strains but not in others (10%). EPEC serotypes or LA was found in 60% (n = 30) of the outbreak strains. The remaining E. coli strains may represent nonpathogenic normal flora, as-yet-undefined pathogens, or pathogens that have lost virulence-associated traits during storage or subculturing.
Nontypeable bacteriophage patterns of methicillin-resistant Staphylococcus aureus involved in a hospital outbreak
Authors: Khalifa K I , Heiba A A , Hancock G
Published: January 2020

Of 93 strains of Staphylococcus aureus isolated from inpatient wards of Ismailia General Hospital, 48 (51%) were proven to be methicillin resistant (MR). Of these MR S. aureus strains, 44 were isolated from patients and 4 were isolated from healthy carriers, who were newly arrived interns working in the same wards. Bacteriophage patterns of MR S. aureus were identified by using routine test dilution (RTD) and 100-fold dilutions (100 RTD) of phages. Of these 48 strains, 37 (75%) (33 from patients and 4 from interns) were nontypeable when using RTD and 100 RTD of phages. Of the other 11 strains, 8 were nontypeable by RTD of phages, but 5 of them had the phage pattern D11/1136 when tested by 100 RTD. Three strains had the phage pattern 3A/3C/55/71, and three strains had different phage patterns, 29/81, 96, and 95/D11. The finding of colonization with virulent MR S. aureus strains in interns working on the wards in which these patients were located suggested that new strategies for control of MR S. aureus nosocomial infections must be considered and evaluated.
Detection of leptospires in urine by polymerase chain reaction
Authors: Van Eys G J , Gravekamp C , Gerritsen M J , Quint W , Cornelissen M T , Schegget J T , Terpstra W J
Published: January 2020

Primers for polymerase chain reaction (PCR) were synthesized from clones derived from a Leptospira hardjo (type hardjobovis) library. One pair of synthetic oligonucleotide primers was selected for further analysis. Under experimental conditions an amplification was obtained with DNA of Leptospira interrogans of some serovars belonging to serogroup sejroe. However, very little or no amplification was observed with DNA from other serovars of this group. No amplification was observed with DNA from other serogroups, other bacteria, or eucaryotic organisms. Cattle urine, seeded with hardjobovis, was processed in several ways and subsequently subjected to PCR. Boiling of the samples or treatment with detergents appeared to be most effective. Urine samples containing fewer than 10 leptospires gave a positive result in the PCR assay. Twenty urine samples obtained from a slaughterhouse or farm cows were investigated using the PCR assay, culture isolation, dot and quick blot hybridization, and serological tests. This comparative study suggests that amplification by PCR may be a valuable method for the detection of leptospires in cattle urine.
Comparison of four commercial brucella agar media for growth of anaerobic organisms
Authors: Mangels J I , Douglas B P
Published: January 2020

Four different commercial brucella blood agar plating media (Anaerobe Systems, BBL Microbiology Systems, Remel, and Scott Laboratories) were compared for the abilities to recover anaerobic organisms from clinical specimens and to support the growth of American Type Culture Collection anaerobic stock cultures. Following 24 h of incubation in an anaerobe chamber, Anaerobe Systems prereduced, anaerobically sterilized brucella plates yielded 63% of the total clinical anaerobe isolates, the Scott medium yielded 51%, the Remel medium yielded 42%, and the BBL medium yielded 37%. Poor growth of Peptostreptococcus magnus, P. anaerobius, Fusobacterium necrophorum, F. nucleatum, and pigmented Bacteroides spp. was observed on brucella media obtained from BBL, Remel, and Scott. Data obtained with stock anaerobic cultures showed that Anaerobe Systems plates yielded good growth and produced a larger colony size with all of the strains tested in 1 day, whereas poor growth of Peptostreptococcus spp., B. melaninogenicus, and Fusobacterium spp. was noted on brucella media from BBL, Remel, and Scott.
New method for determination of efficacy of health care personnel hand wash products
Authors: Mahl M C
Published: January 2020

A method of studying the effects of health care personnel hand wash products is described. The fingernail regions of the hands of volunteers are inoculated with a mixture of Escherichia coli and Serratia marcescens, and the areas are dried for a standard time. After routine hand washing, each fingernail region is individually scrubbed with an electric toothbrush which moves longitudinally to the handle into collection fluid contained in a petri dish. The test bacteria in the fluid are then enumerated. (Bacillus subtilis spores may be included as tracers to show degree of physical removal of the procedure.) This method has several advantages over the frequently used glove juice technique. Experimental designs with large numbers of volunteers, multiple sampling sites, and many hand wash products may be performed. Ten sampling sites (fingers) are available, versus the two gloved hands for testing products. (Efficiency is almost 100% in the recovery of spore tracers placed on the fingernails.) Many commercial health care personnel hand wash products containing antimicrobial agents substantive to the skin do not rapidly reduce numbers of inoculated bacteria in the fingernail regions to any greater extent than nonantimicrobial hand washes. Products containing isopropanol or ethanol are very effective in decreasing bacteria in areas around and under the fingernails.
Visuwell Reagin, a non-treponemal enzyme-linked immunosorbent assay for the serodiagnosis of syphilis
Authors: White T J , Fuller S A
Published: January 2020

A urease-based enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of reagin antibodies in serum. Visuwell Reagin (ADI Diagnostics Inc., Rexdale, Ontario, Canada) is a non-treponemal screening test for the serodiagnosis of syphilis which has the benefits of large batch testing, automatability, and objective interpretation of results. Unheated, undiluted sera are incubated in 96-well microtiter plates coated with a modified cardiolipin-lecithin-cholesterol antigen. Antibody bound to the plate is detected by an anti-human immunoglobulin G-urease conjugate. The procedure consists of three steps, with a total test time of 60 min. Visuwell Reagin ELISA was compared with the Venereal Disease Research Laboratory (VDRL) test and the reagin screening test (RST) with the following results. For ELISA versus the VDRL test, the sensitivities for untreated syphilis (n = 37) were 97.3% for both ELISA and the VDRL test, the confirmatory positive values (n = 79) were 84.8% for ELISA and 72.2% for the VDRL test, and the specificities for normal samples (n = 1,327) were 98.8% for ELISA and 99.5% for the VDRL test. For ELISA versus RST, the sensitivities for untreated syphilis (n = 57) were 94.7% for ELISA and 87.7% for RST, the confirmatory positive values (n = 26) were 96.2% for ELISA and 92.3% for RST, and the specificities for normal samples (n = 1,891) were 99.6% for ELISA and 99.3% for RST. The overall concordance values of ELISA with VDRL test and RST were 96.7 and 97.9%, respectively. The specificity of ELISA compared with that of RST may be underestimated, since confirmatory data were not available for all apparent false-positive samples. Visuwell Reagin had increased sensitivity and similar specificity compared with flocculation tests.
Lactobacillus acidophilus inhibits growth of Campylobacter pylori in vitro
Authors: Bhatia S J , Kochar N , Abraham P , Nair N G , Mehta A P
Published: January 2020

Campylobacter pylori has been implicated as a causative factor in acid-peptic disease. Lactobacillus acidophilus is known to inhibit the growth of pathogens in the human gastrointestinal tract. We recovered C. pylori from gastric antral biopsies of seven patients with acid-peptic disease; the isolates were then cultured in brucella broth. The effect of L. acidophilus (cultured in DeMan-Rogosa-Sharpe broth) on the growth of C. pylori was tested by a mixed culture technique. L. acidophilus inhibited the growth of all seven isolates of C. pylori in vitro. All these isolates were also inhibited by the L. acidophilus culture supernatant (brucella blood agar cup technique) obtained at or after 48 h of incubation. Inhibition of C. pylori growth was also observed with 1 and 3% lactic acid but not with 0.5 and 1% hydrogen peroxide, the L. acidophilus sonic extract, or a citrate-phosphate buffer (pH 4.0). We conclude that the inhibitory action of L. acidophilus on C. pylori is dependent on an extracellular secretory product, probably lactic acid. This inhibitory effect may be of therapeutic relevance in patients with C. pylori-positive acid-peptic disease.
Laboratory and clinical evaluation of a commercial DNA probe for the detection of Legionella spp
Authors: Pasculle A W , Veto G E , Krystofiak S , McKelvey K , Vrsalovic K
Published: January 2020

We prospectively compared a commercially available Legionella DNA probe with culture and direct immunofluorescence. The analytical sensitivities of the DNA probe and direct immunofluorescence were equal. Both tests detected 4 X 10(3) CFU of Legionella pneumophila or Legionella micdadei per ml in the pulmonary secretions of experimentally infected guinea pigs. The diagnostic sensitivity of the reagent was evaluated by using 809 samples of respiratory secretions. Of 51 DNA probe-positive specimens, 31 came from patients with culture-confirmed legionellosis. Two culture-positive specimens had negative DNA probe tests. The sensitivity and specificity of the DNA probe were 93.9 and 97.4%, respectively. The sensitivity and specificity of direct immunofluorescence were 68.9 and 99.6%, respectively. The low specificity of the DNA probe resulted in an unacceptable positive predictive value (60.8%). False-positive DNA probe tests were not due to nonspecific binding of the probe or to technical problems but were associated with one lot of probe reagent. Most of the false-positive probe tests had values near the threshold value of greater than or equal to 4.0 suggested by the manufacturer. Raising the threshold value for a positive test to 7 lowered the sensitivity to 69.2% but raised the specificity to 99.2%. At this level, the performances of the DNA probe and direct fluorescent-antibody testing were equivalent. Respiratory secretions from patients receiving therapy for culture-confirmed Legionella infection remained DNA probe positive for up to 8 days, even though cultures and/or direct immunofluorescence tests often became negative. The DNA probe test is a satisfactory replacement for direct immunofluorescence but cannot replace culture for the laboratory diagnosis of Legionella infections.
Aerobic and anaerobic microbiology of biliary tract disease
Authors: Brook I
Published: January 2020

A retrospective analysis of the experiences of two military hospitals over 4 years in the recovery of organisms from biliary tract specimens was done. Bacterial growth was obtained in 123 bile specimens. Aerobic and facultative bacteria only were present in 59 specimens (48%), aerobic bacteria only were present in 4 specimens (3%), and mixed anaerobic and aerobic or facultative bacteria were present in 60 specimens (49%). Of 286 isolates recovered, 216 were aerobic or facultative (1.8 per specimen) and 70 were anaerobic (0.6 per specimen). The predominant bacteria were Escherichia coli (71 isolates), group D streptococci (42 isolates), Klebsiella sp. (29 isolates), Clostridium sp. (27 isolates), Bacteroides sp. (28 isolates), and Enterobacter sp. (16 isolates). Polymicrobial infections were present in 108 instances (88%). A higher recovery rate of anaerobes was present in patients with chronic infections than in those with acute infections and did not correlate with the presence of gallstones or use of antimicrobial prophylaxis.
Detection of antibodies to human immunodeficiency virus type 1 in whole blood and saliva by using a passive hemagglutination test
Authors: Vasudevachari M B , Uffelman K , Pierce P F , Lane H C , Salzman N P
Published: January 2020

A passive hemagglutination test (PHA) for detecting human immunodeficiency virus type 1 antibodies in serum samples by using envelope glycoprotein (gp160)-coupled sheep erythrocytes was described earlier (M.B. Vasudevachari, K. U. Uffelman, T.C. Mast, R.L. Dewar, V. Natarajan, H.C. Lane, and N.P. Salzman, J. Clin. Microbiol. 27:179-181, 1989). In the study reported here, the applicability of the PHA test to the detection of antibodies in whole-blood and saliva samples has been investigated. We observed a 100% correlation between PHA and commercial enzyme-linked immunosorbent assay in 101 whole-blood samples and 98% correlation between PHA and reactivity to envelope proteins in Western blots (immunoblots) of 53 saliva samples. Furthermore, salivary antibodies could be detected in 19 of the 22 seropositive individuals. As in serum, antibodies to envelope proteins were widely prevalent in all the Western blot-reactive saliva samples.
Assay of heat-labile enterotoxins by their ADP-ribosyltransferase activities
Authors: Narayanan J , Hartman P A , Graves D J
Published: January 2020

Simple enzymatic assays to detect heat-labile enterotoxins whose modes of action are similar to that of cholera toxin were evaluated. The assays are performed by using an artificial substrate, diethylamino benzylidine-aminoguanidine, which is an ADP-ribose acceptor. The product, formed in the presence of NAD+, can be quantitated by spectrofluorometric, spectrophotometric, or high-performance liquid chromatographic (HPLC) methods. As little as 25 ng (spectrofluorometry) or 125 ng (spectrophotometry or HPLC) of cholera toxin can be detected in an assay volume of 250 microliters. The detection limit for heat-labile enterotoxin by either the spectrophotometric or HPLC methods was 125 ng/250 microliters. Because the results are quantitative, the enzymatic methods can be used for medium development, determination of factors that influence toxin production, and other applications that heretofore could be accomplished only with difficulty. The enzymatic methods add a new dimension to the assay of toxins that ribosylate arginine residues of proteins. Sensitivities of the assays might be improved by developing better synthetic substrates, and applications could be broadened by the development of artificial substrates containing other functional groups.
Single-source outbreak of Candida tropicalis complicating coronary bypass surgery
Authors: Isenberg H D , Tucci V , Cintron F , Singer C , Weinstein G S , Tyras D H
Published: January 2020

Candida tropicalis was isolated from the sternal wounds of eight coronary bypass patients from 18 to 89 days postoperatively; infections were limited to soft tissue in five patients but involved the sternum in three patients. Analysis of surgery records implicated one individual as the potential source of the yeast; this was confirmed by microbiological studies of fingertips and nasopharynx cultures of all personnel in contact with these patients. Only the suspect nurse, then acting as a scrub nurse and not as a circulator, infected the eight patients. Her removal from the cardiac team terminated the cluster outbreak.
Direct detection of cytomegalovirus from bronchoalveolar lavage samples by using a rapid in situ DNA hybridization assay
Authors: Gleaves C A , Myerson D , Bowden R A , Hackman R C , Meyers J D
Published: January 2020

An in situ DNA hybridization assay was compared with centrifugation culture for rapid detection of cytomegalovirus (CMV) from bronchoalveolar lavage (BAL) samples. Eighty BAL samples were inoculated into both centrifugation culture and standard culture. Cytospin preparations of the BAL samples were studied in a 75-min in situ DNA hybridization assay using the PathoGene CMV kit (Enzo Biochem, Inc., New York, N.Y.). Of the 80 samples, 39 (49%) were positive for CMV; 37 of 39 (95%) were positive by centrifugation culture, 34 of 39 (87%) were positive in standard culture, 24 of 39 (62%) were positive by in situ hybridization, and 20 of 39 (56%) were positive by histologic and/or immunofluorescence techniques. The in situ hybridization assay detected 23 of the 37 samples positive in centrifugation culture, for a sensitivity of 62% and a specificity of 98%. We conclude that the in situ hybridization assay is a specific and more rapid test than centrifugation culture and standard culture for diagnosis of CMV pulmonary infection. For the clinical laboratory, however, current hybridization methods are not sufficiently sensitive to replace centrifugation culture for detection of CMV in BAL specimens.
Comparative evaluation of radiometric tryptic soy broth versus radiometric tryptic soy broth with 10% sucrose for detection of bacteremia and fungemia in pediatric patients
Authors: Weinstein M P , Reller L B , Mirrett S , Stratton C W , Paisley J W , Lauer B A
Published: January 2020

We compared BACTEC radiometric blood culture media with (8B) and without (6B) 10% sucrose for the detection of bacteremia and fungemia in pediatric patients at four university teaching hospitals that used identical methods for obtaining and processing specimens. Overall, the yields of microorganisms from 5,714 blood culture sets were no different in the two media, although a trend was noted favoring 6B for the detection of pneumococci. Speed of detection of positive results was faster in the 6B than in the 8B medium (P less than 0.05), largely due to the faster detection of Staphylococcus aureus in the 6B medium. We conclude that, overall, with pediatric patients the hypertonic 8B radiometric medium has no advantage and that it possibly has a modest disadvantage, compared with isotonic 6B radiometric medium.
Evaluation of the Merifluor-Legionella immunofluorescent reagent for identifying and detecting 21 Legionella species
Authors: Edelstein P H , Edelstein M A
Published: January 2020

We evaluated a 33-valent polyclonal indirect immunofluorescent-reagent kit (Merifluor-Legionella; Meridian Diagnostics Inc., Cincinnati, Ohio) made for the detection of Legionella species by testing bacterial isolates, seeded sputum, and negative sputum samples. Use of the reagent according to the directions of the manufacturer gave false-negative staining of homologous culture isolates due to a prozone phenomenon; this was solved by diluting test strain suspensions. After this change in testing protocol was made, the reagent gave bright fluorescent staining with 31 of the 33 Legionella strains with which it supposedly reacts. Strongly reacting Legionella strains included the type strains of L. pneumophila serogroups 1 to 10, L. longbeachae serogroups 1 and 2, and serogroup 1 of L. anisa, L. bozemanii, L. cherrii, L. dumoffii, L. gormanii, L. hackeliae, L. jamestowniensis, L. jordanis, L. maceachernii, L. micadedi, L. oakridgensis, L. rubrilucens, L. sainthelensi, L. spiritensis, L. steigerwaltii, and L. wadsworthii. Type strains of L. erythra and L. feeleii fluoresced only dimly with the reagent. Of 10 non-Legionella bacteria known to cross-stain with other polyvalent antisera, 5 also cross-reacted with the Merifluor reagent; these included 3 Bacteroides fragilis and 2 Pseudomonas fluorescens strains. The lower limit of detection of L. pneumophila serogroup 1 in seeded sputum was about 5 x 10(4) to 5 x 10(5) cells per ml. None of 21 randomly collected sputum specimens tested contained fluorescing legionellalike organisms, but 6 specimens did contain brightly fluorescing bacteria atypical in morphology for Legionella species. The Merifluor-Legionella kit appears to perform as well as other polyclonal immunofluorescent reagents used for detection of Legionella species. Because of the cross-reactions observed, which are common to all polyclonal reagents, utilization of this reagent for either bacterial identification or detection must be performed in combination with culture.
New selective medium for the isolation of Neisseria gonorrhoeae
Authors: Evans G L , Kopyta D L , Crouse K
Published: January 2020

GC-Lect, a new selective medium for the isolation of Neisseria gonorrhoeae which contains five antimicrobial agents, was evaluated with stock cultures and with 500 clinical specimens. With stock cultures, vancomycin-resistant Staphylococcus epidermidis that grew on modified Thayer-Martin medium (MTM) was inhibited on the new medium. Also, vancomycin-susceptible strains of N. gonorrhoeae were much less inhibited on the new medium than on Martin-Lewis agar or MTM. With oropharyngeal cultures of healthy volunteers, Capnocytophaga species were frequently isolated on MTM from two of three manufacturers but were completely inhibited on GC-Lect. In the clinical study, visible growth of N. gonorrhoeae occurred within 24 h in 72% of the positive cultures on GC-Lect, compared with only 52% on the reference medium. A total of 50 positive cultures were obtained with GC-Lect, compared with 49 obtained with MTM. The selectivity of GC-Lect was superior, with only 19 cultures producing growth of normal flora, compared with 78 cultures on MTM after 24 h of incubation. The selectivity was especially improved on GC-Lect with regard to yeasts (2 versus 30 cultures) and gram-positive cocci (5 versus 31 cultures).
Reassessment of susceptibility test interpretive criteria for ticarcillin and ticarcillin-clavulanic acid
Authors: Fuchs P C , Jones R N , Barry A L
Published: January 2020

There are at least four different existing or proposed interpretive criteria for the disk diffusion susceptibility testing of ticarcillin and ticarcillin plus clavulanic acid (T/C). To assess these criteria, 570 gram-negative bacillary isolates were tested for susceptibility to ticarcillin and T/C by both disk diffusion and broth microdilution methods. These included 53 strains of the family Enterobacteriaceae selected for ticarcillin resistance and high-level beta-lactamase production. The broth microdilution test results were more influenced by increased beta-lactamase production than were disk diffusion results. In the absence of published data indicating which of the two standardized test methods better predicts clinical response, we conclude that until such data are available the more conservative National Committee for Clinical Laboratory Standards tentative criteria for tests with members of the Enterobacteriaceae are appropriate. Our data do not support the use of separate T/C interpretive criteria for Pseudomonas spp. and members of the Enterobacteriaceae. The appropriateness of different interpretive criteria needs further evaluation.
Isolation of a Campylobacter-like organism from healthy Syrian hamsters (Mesocricetus auratus)
Authors: Stills H F , Hook R R , Kinden D A
Published: January 2020

A Campylobacter-like organism was isolated from the ilea of normal hamsters. The organism was isolated from an ileal homogenate which was passed through a filter (0.65-micron pore size) and cultured on blood-agar plates in a microaerophilic atmosphere at 37 degrees C. Pinpoint translucent colonies were first observed after 120 h of incubation. The isolated organism measured 2.0 to 3.5 microns in length (excluding flagella) by 0.17 to 0.25 micron in width and typically had a single terminal sheathed flagellum. The organism was oxidase, catalase, and urease positive, reduced nitrates, and was susceptible to nalidixic acid (30-micrograms disk) and resistant to cephalothin (30-micrograms disk). Unlike Campylobacter pylori subsp. mustelae, this organism did not hydrolyze indoxylacetate. Immunofluorescence studies with a Campylobacter species-specific monoclonal antibody (8322-2E6) revealed the presence of numerous positively stained organisms within the crypt epithelial cells of the hamsters from which this organism was isolated. The role of this organism in the pathogenesis of proliferative ileitis in hamsters is uncertain, as is the taxonomic relationship of this organism to other members of the genus Campylobacter.
Accuracy and reproducibility of the 4-hour ATB 32A method for anaerobe identification
Authors: Kitch T T , Appelbaum P C
Published: January 2020

The ATB 32A system (API System SA, La Balme les Grottes, Montalieu-Vercieu, France) was evaluated for use in the identification of 214 anaerobes. Organisms included 73 isolates of the Bacteroides fragilis group, 24 Bacteroides spp., 10 fusobacteria, 43 clostridia, 28 cocci, and 36 gram-positive, nonsporeforming rods. With the concomitant use of Gram stain, pigmentation, catalase testing, and aerobic growth, the ATB 32A system correctly identified 97% of the B. fragilis group isolates, 88% of Bacteroides spp., 50% of fusobacteria, 74% of clostridia, 100% of cocci, and 86% of the gram-positive, nonsporeforming rods. Overall, 188 strains (88%) were correctly identified, with 18 (8%) requiring extra tests, other than the four mentioned above, for correct identification. Eight strains were misidentified, including one Bacteroides sp., three fusobacteria, one Clostridium sp., and three gram-positive, nonsporeforming rods. Reproducibility was very good, with 12 of 14 strains (86%) tested in triplicate yielding identical correct results on each of three occasions and 2 strains (14%) yielding identical correct results on two occasions. There was a low-probability identification (including the correct species) on the third testing. The ATB 32A system represents a worthwhile advance in systems used for the identification of clinically significant anaerobic bacteria.