bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical comparison of aerobic, hypertonic, and anaerobic culture media for the radiometric detection of bacteremia
Authors: Coleman R M , Laslie W W , Lambe D W
Published: May 2021

The BACTEC 225 was used to test 5,811 routine blood cultures over a 20-month period. Aerobic, anaerobic, and hypertonic media were employed. The BACTEC 225 detected 511 positive cultures; 407 of these were considered significant organisms, and 104 were presumed contaminants. Of the significant positive cultures, 15% were detected within the first 12 h of incubation, 52% within 24 h, 82% within 48 h and 92% within 72 h. Aerobic, anaerobic, and hypertonic media are recommended for each venipuncture since 56 cultures were isolated from the aerobic medium only, 110 from the anaerobic medium only, and 94 from the hypertonic medium only. There were 16 patients who had multiple venipunctures from which organisms were repeatedly isolated from only one medium: two from the aerobic medium, four from the anaerobic medium, and ten from the hypertonic medium only. Detection times were not significantly different for the aerobic and hypertonic media. However, there were five patients with multiple venipunctures in which growth was detected radiometrically at least 48 h earlier in the hypertonic than in the aerobic medium. False-positive growth index readings were noted in 1,085 (19%) of the aerobic vial, 11 (0.19%) of the anaerobic vials, microorganisms were isolated from at least one of the companion vials. Using 5% co2 to flush the aerobic vials. With some false-positive aerobic and hypertonic vials decreased the number of false positives to about 6% of the total.
Apparent false positive histoplasmin latex agglutination tests in patients with tuberculosis
Authors: DiSalvo A F , Corbett D S
Published: May 2021

Serum specimens from patients admitted to a respiratory disease hospital were examined by the histoplasmin latex agglutination, the complement fixation, and the agar gel immunodiffusion tests. Of 300 sera examined, 21 (7.0%) gave an apparent false positive reaction at a dilution of 1:16 or greater. Fourteen (66%) of the 21 patients studied has culturally proven tuberculosis. One patient each had a diagnosis of hypertensive cardiovascular disease with congestive heart failure, infection with atypical mycobacteria (Runyon group III), chronic pneumonitis secondary to gunshot wound, and pulmonary abscess of unknown etiology; two had bronchogenic carcinoma; and one serum specimen came from an apparently healthy employee. The results of the histoplasmin latex agglutination test should be interpreted with caution, particularly if only one serological determination has been made and the titer is low.
Use of phage F-phi WJ-1 of Mycobacterium fortuitum to discern more phage types of Mycobacterium tuberculosis
Authors: Jones W D , Greenberg J
Published: May 2021

A total of 125 strains of Mycobacterium tuberculosis from the Southeastern area of the United States was subjected to phage typing. In addition to the five major mycobacteriophages, a new phage, F-phi WJ-1, was used in the study. The results obtained with the five major phages were: type A0, 35.2%; TYPE B, 29.6%, and type C, 4.0%. The remaining 21.2% of the strains phaged typed as subgroups A1 through A6. These percentages were similar to the typing results of earlier studies. The new phage, F-phi WJ-1, subdivided each of the phage types, with the exception of type C, into two subgroups. The possible role of host modification-restriction of the phages used in phage typing of strains of M. tuberculosis is discussed.
Effect of types of media on the production of acid from glucose by so-called glucose-negative strains of Neisseria gonorrhoeae
Authors: Baron E S , Saz A K
Published: May 2021

Typical gonococci metabolize glucose; however, occasional strains of Neisseria gonorrhoeae fail to metabolize glucose when tested on cystine Trypticase agar (CTA) medium, a fact that leads to delay in identification. Certain strains of so-called glucose-negative N. gonorrhoeae do indeed metabolize glucose, depending on the medium used in testing for metabolism of the carbohydrate. Six strains were tested that failed to oxidize glucose with the production of acid when tested on standard CTA medium, yet all produced acid from glucose when supplemented GC medium with a phenol red indicator was utilized. An attempt was made to single out the compound present in CTA that leads to inhibition of metabolism and, occasionally, growth as well. We found that certain ratios of the cystine and Na2SO3 concentrations are inhibitory, including that ratio of the two compounds present in CTA medium; however, L-cysteine, when included in similar concentrations, did not inhibit the metabolic reaction.
Method for identifying Salmonella and Shigella directly from the primary isolation plate by coagglutination of protein A-containing staphylococci sensitized with specific antibody
Authors: Edwards E A , Hilderbrand R L
Published: May 2021

A technique is described that allows presumptive identification of either Salmonella or Shigella organisms directly upon the original isolation plate, in this case, MacConkey agar. This was accomplished by applying a drop of specifically sensitized protein A-containing Staphylococcus aureus over a "suspected" colony or several colonies of organisms grown on MacConkey agar. The plate is tilted to and fro to allow mixing of the particles with specific antigen that ir readily solubilized from the colony and observing for agglutination of the sensitized particles by use of a dissecting microscope. The agglutination can frequently be seen within 15 s, increasing in intensity over a 2-min period. The polyvalent Salmonella antiserum was slower in developing strong agglutination (1.5 to 2 min) compared to particles sensitized with group-specific antisera (15 to 45 s). A high-titer antiserum was important for a test reagent to have the required sensitivity.
Inhibition of Bacteroides fragilis on blood agar plates and reversal of inhibition by added hemin
Authors: Wilkins T D , Chalgren S L , Jimenez-Ulate F , Drake C R , Johnson J L
Published: May 2021

Bacteroides fragilis strains formed much smaller colonies on most types of blood agar plates than they did on the same media without blood. Blood inhibited strains of B. fragilis subsp. distasonis the most and B. fragilis subsp. fragilis the least. The inhibition could be eliminated by adding hemin to the blood agar. The inhibitory component of the blood was inside the erythrocytes and appeared to be the hemin-free globin of hemoglobin.
Detection of neonatal calf diarrhea virus, infant reovirus-like diarrhea virus, and a coronavirus using the fluorescent virus precipitin test
Authors: Peterson M W , Spendlove R S , Smart R A
Published: May 2021

Thirty-four calf and five infant fecal specimens were tested for the neonatal calf diarrhea virus (NCDV) and for the reovirus-like infantile diarrhea agent; respectively. The procedures used were the fluorescent virus precipitin test and immune electron microscopy. Fourteen of the calf stools contained detectable NCDV, and four of the five infant stools contained the reovirus-like human agent. Infectious NCDV was detected in four of the 34 calf fecal specimens when Madin-Darby bovine kidney cell cultures that had been inoculated with supernatant fluids from stool suspensions were stained with fluorescent antibody. The 20 calf stools that did not have detectable virus were examined for the bovine corona diarrhea virus. Coronavirus was found in two of these specimens.
Inhibition of Streptococcus mutans strains by different mitis-salivarius agar preparations
Authors: Staat R H
Published: May 2021

Several Streptococcus mutans strains were markedly inhibited by mitis-salivarius agar manufactured by Baltimore Biological Laboratories, but little, if any, inhibition was noted using Difco Laboratories' mitis-salivarius agar. Supplementation of the basic medium with sucrose and bacitracin for specific selection of S. mutans resulted in suppression of representative S. mutans type a strains regardless of manufacturer.
Pitting of Agar Surface by
Pseudomonas stutzeri
Authors: Alexander Jane J. , Lewis Jay F.
Published: May 2021

A strain of
Pseudomonas stutzeri
which exhibited pitting of agar surfaces is described.
Evaluation of a Proteus rettgeri O-serotyping system for epidemiological investigation
Authors: Penner J L , Hinton N A , Hennessy J N
Published: May 2021

An evaluation of the serotyping system for Proteus rettgeri has been performed through its application to 561 human isolates of this species. Of these, 545 were typable and could be separated into 58 O serotypes. A group of isolates from one hospital, and clinically implicated in cross-infections, was found to belong the same O serotype, confirming the clinical findings. Isolates with more than one specificity of thermostable antigen, in most cases, were found to have combinations of such specificities restricted to combinations previously defined for the O-type strains. Furthermore, the use of pooled antisera was investigated and system of pooling was devised to retain the specificity of the system and to increase the facility of the typing procedure.
Citrobacter diversus isolated from clinical material
Authors: Altmann G , Sechter I , Cahan D , Gerichter C B
Published: May 2021

Forty-seven strains of Citrobacter diversus were isolated during a 12-month period from clinical material obtained from patients in a general hospital in Israel. The majority of cultures (38) were recovered from urine and wound discharges. There was one case of septicemia. The biochemical reactions of all cultures were typical for this species, except one that was anaerogenic, and they could be divided into five biotypes (a through e). Thirty-nine strains were identified serologically and found to belong to one of seven O groups described by Gross and Rowe (1974). All cultures were sensitive to tetracycline and nalidixic acid. All strains were resistant to carbenicillin and ampicillin and produced beta-lactamase.
Application of a gloved-hand model for multiparameter measurements of skin-degerming activity
Authors: Michaud R N , McGrath M B , Goss W A
Published: May 2021

The application of an established gloved-hand model to multiparameter measurements of skin-degerming activity is described. In particular, appropriate experimental designs are illustrated which allow characterization of performance of topical skin-cleansing preparations in terms of rapid, sustained, cummulative, and persistent skin-degerming effects on the hand. Single-contact studies were used to define the degerming activity profiles of selected commercial surgical scrub preparations, and to establish the optimal post-treatment sampling interval for individual preparations. Rapid and sustained skin-degerming effects were measured and contrasted. Rapid skin-degerming activity, iodophor preparation. Sustained skin-degerming activity, namely, that occurring on the gloved hand during a postcontact interval, was shown and characterized for two hexachlorophene preparations. Multiple-contact studies with a 3% hexachlorophene preparations were used to illustrate cummulative and persistent skin-degerming effects. Cummulative skin-degerming activity was demonstrated in terms of progressive bacterial reductions after repeated contacts within a single day. Presistent skin-degerming activity was shown in terms of the profile of daily pretreatment bacterial counts after multiple contacts over successive days. Uniformity of treatment response was established for a broad range of pretreatment bacterial counts extending from approximatley log 4 to log 7 per hand. The importance of pretreatment bacterial count measurement and of adequate neutralization of hand extract samples is stressed. A randomized-hand experimental design is discussed relative to its versatility and amenability to statistical analysis.
Clinical isolates of anaerobic gram-negative rods with a formate-fumarate energy metabolism: Bacteroides corrodens, Vibrio succionogenes, and unidentified strains
Authors: Smibert R M , Holdeman L V
Published: May 2021

Strains of anaerobic, gram-negative bacteria, isolated from human clinical specimens and from studies of human normal flora, that have energy metabolism similar to Vibrio succinogenes are described. Included are four human isolates of V. succinogenes, five similar strains of motile straight rods, three strains of Bacteroides corrodens, and two unidentified strains. All strains studied grew poorly in usual anaerobic broth media but produced good turbidity in overnight broth cultures in media containing fromate and fumarate, indicating that all have an energy metabolism similar to V. succinogenes: they gain energy by the transfer of electrons from formate or hydrogen to fumarate.
Fluorescent antibody test for the serological diagnosis of gonorrhea
Authors: Gaafar H A , D'Arcangelis D C
Published: May 2021

An indirect fluorescent antibody technique has been developed for the serological diagnosis of gonorrhea. The selected strain(s) of Neisseria gonorrhoeae possesses a heat-labile surface antigen (L-antigen). Sera are diluted 1:10, and an aliquot is hear inactivated at 59 C for 30 min. The treated and untreated aliquots are then examined for human immunoglobulin G anti-L-antigen. In a prelimiary study of 495 sera, 95% of those from women with a bacteriologically confirmed diagnosis of gorrhea and 87% of those from male patients were reactive in this test, whereas only 1.4% fo the sera from presumably normal individuals were reactive.
Mycobacterium resembling Mycobacterium fortuitum that produces brown pigment
Authors: Hawkins J E , Falco E B
Published: May 2021

Two cultures of acid-fast bacilli with characteristics most closely resembling those of Mycobacterium fortuitum were recovered as casual isolates from sputa of a patient with an apparent brochogenic tumor. One of the cultures was consistenly cream colored to rosy buff. The other, however, changed from buff to rust to dark brown and had the gross appearance of a fungus culture.
Radioimmunoassay for measuring antibodies specific for group B streptococcal types Ia, Ib, Ic, II, and III
Authors: Wilkinson H W , Jones W L
Published: May 2021

The object of this study was to develop a test that would measure antibodies directed against group B streptococcal types Ia, Ib, Ic, II, and III. The type-specific carbohydrate antigens were purified, labeled with 125I, and used to develop a radioimmunoassay. This procedure should be particularly useful in testing human sera for group B type-specific antibodies, since it requires very small quantities of antigens and measures primary type antigen-antibody interactions.
Enzyme-linked immunosorbent assay for streptococcal M protein antibodies
Authors: Russell H , Facklam R R , Edwards L R
Published: May 2021

The enzyme-linked immunosorbent assay (ELISA) described by Engvall and Perlmann, which uses antigen-coated tubes and enzyme-labeled anti-immunoglobulin, has been used for the detection of antibodies against streptococcal M protein. The antigen used in the assay was obtained by guanidine extraction of type M-12 streptococcal cell walls followed by hydroxyapatite chromatography. This antigen has the capacity to elicit bactericidal antibodies in rabbits. The results show that the ELISA is specific and highly sensitive for the detection of antibodies in rabbit and human antisera. Preliminary results suggest that, when M-12 antigen is used, the antibodies detected by ELISA are the same antibodies detected in the bactericidal test. The assay has been performed with human and rabbit sera. There was a 96% agreement between bactericidal and ELISA results with rabbit sera and 97.5% agreement with human sera. All bactericidal antibody-positive sera tested thus far yielded positive ELISA results.
Description of a polyvalent conjugate and a new serogroup of Bacteroides melaninogenicus by fluorescent antibody staining
Authors: Lambe D W , Jerris R C
Published: May 2021

A polyvalent conjugate (fluorescein isothiocyanate-labeled antibody reagent) containing serogroups A, B, and C conjugates was prepared. This polyvalent conjugate gave a positive fluorescent antibody (FA) stain with 49 stains of Bacteroides melaninogenicus representing serogroups A, B, and C. When additional strains (92 strains) of the three subspecies of B. melaninogenicus were examined by the FA stain, with A, B, and C, and polyvalent conjugates, nine strains of B. melaninogenicus subsp. intermedius failed to give a positive stain with any conjugate. Therefore, an FA conjugate was prepared with the antiserum to one of these strains (532-70A); all nine strains stained positively with this conjugate. These nine strains were biochemically characteristic of B. melaninogenicus subsp. intermedius; thus, these strains were designated as a new serogroup, serogroup C-1. A new polyvalent conjugate containing serogroups A, B, C, and C-1 was prepared. This polyvalent conjugate stained positively with 23 representative strains from serogroups A, B, C, and C-1. The new conjugates failed to stain positively with other anaerobes and aerobes tested. The four individual conjugates, as well as the polyvalent conjugate, may be used for a more rapid identification of B. melaninogenicus than is possible by biochemical testing.
Phage typing of Staphylococcus epidermidis
Authors: Talbot H W , Parisi J T
Published: May 2021

Thirteen phages were isolated from lysogenic cultures of Staphylococcus epidermidis from a clinical laboratory and used to type 223 clinical isolates of this organism. The 18 phages isolated in The Netherlands were used to type these same cultures. No correlation was observed between phage type, biotype, or clinical source of isolation. At phage concentrations of 100 times the routine test dilution, 35.0% of the cultures were typable with out phages and 21.5% were typable with the phages from The Netherlands. When only cultures in biotype 1 were considered, 43.3 and 24.1% of 141 cultures were typable with our phages and those from The Netherlands, respectively. The lytic reactions obtained with our phages were generally stronger and easier to read and the lytic patterns were, almost invariably, shorter. The typability of untypable cultures was increased 12.0% by incubation at 45 C prior to phage typing and 20% by heat shock (55 C for 5 min) prior to typing. Phage typing 5 subcultures of 20 typable cultures on 5 successive days showed that the lytic patterns were reproducible. The present status of phage typing S. epidermidis and the work needed to obtain a set of typing phages for epidemiological studies of infections by this organism are discussed.
Purification of influenza virus with Arcton 113
Authors: Brady M I , Furminger I G
Published: May 2021

Arcton 113 has proved effective in the removal of host membrane fragments which cosediment with influenza virus during differential centrifugation. The virus appeared morphologically undamaged by the procedure and retained its infectivity, neuraminidase, and hemagglutinating activity. Pyrogenicity, however, was considerably reduced.
Comparison of rhinovirus-sensitive HeLa cells and human embryo fibroblasts for isolation of rhinoviruses from patients with respiratory disease
Authors: Lewis F A , Kennett M L
Published: May 2021

Rhinovirus-sensitive HeLa cells (HeLa "R") and human embryo lung fibroblasts (HEL) were compared for the isolation of rhinoviruses from patients with respiratory disease. In the period May 1970 to December 1974, 526 rhinoviruses were isolated from 517 patients, 32% in both cell types, 59% in HeLa "R" only, and 9% in HEL only. The annual isolations in HeLa "R" were between 1.7 and 4 times greater than in HEL cells and may have been due to changes in the sensitivity of the cells or the prevalence of serotypes favoring the HeLa "R" cells. Acid lability was more easily demonstrated in HeLa "R" than in HEL cells, because the infectivity titers obtained were 10- to 100-fold higher.
Application of a new method for detecting streptococcal nicotinamide adenine dinucleotide glycohydrolase to various M types of Streptoccus pyogenes
Authors: Lütticken R , Lütticken D , Johnson D R , Wannamaker L W
Published: May 2021

The production of extracellular nicotinamide adenine dinucleotide glycohydrolase (NADase) by Streptococcus pyogenes can easily be demonstrated using the fluorescence of nicotinamide adenine dinucleotide in ultraviolet light occurring on addition of strong alkali. The new method described here uses microtiter plates and can be read with the naked eye. It permits the screening of large numbers of strains in a short time and with minimal amounts of reagents. The sensitivity of the new method proved to be good in comparison with the bisulfite spectrophotometric method. Culture supernatants of 177 group A streptococci were tested for NADase production by the microtiter fluorescence method. We could confirm former findings that strains within a certain M type of S. pyogenes are usually either producers or nonproducers of the enzyme. The usefulness of the test for screening of streptococcal NADase production is discussed.
Delayed hypersensitivity cross-reactions between Sporothrix schenckii and Ceratocystis species in sporotrichotic patients
Authors: Ishizaki H , Nakamura Y , Kariya H , Iwatsu T , Wheat R
Published: May 2021

Cutaneous delayed hypersensitivity to antigens prepared from Sporothrix schenckii and several Ceratocystis species, including C. stenoceras, C. ulmi, C. ips, and C. minor, was tested in 14 patients with known cutaneous sporotrichosis. Extensive cross-reactions were observed. Nonsporotrichotic people (controls) did not react to these antigens. The correlation coefficient between antigens of S. schenckii and each Ceratocystis species was calculated from the areas of the cutaneous reactions. Among the Ceratocystis species tested, the correlation coefficient between S. schenckii and C. stenoceras was 0.91.
Serological studies and isolations of serotype hardjo and Leptospira biflexa strains from horses of Argentina
Authors: Myers D M
Published: May 2021

Three pathogenic leptosipras and 12 saprophytic Leptospira biflexa strains were isolated from 72 apparently normal horse kidneys collected at an abattoir in Argentina. Cross-agglutination reaction patterns of the pathogens showed that they were antigenically homologous with members of the Hebdomadis group. When one of the strains was compared to Hebdomadis serotypes in reciprocal agglutination-absorption tests, it was found to be serologically homologous to serotype hardjo. This is the first known report of an isolation of this serotype from horses. Serological tests were also carried out on randomly collected abattoir sera from 245 horses to determine the prevalence of equine leptospirosis. Significant antibody titers (1:100 or greater) were found in 74.6% of the sera. Predominant reactions occurred with the antigens pomona, hebdomadis group, pyrogenes, tarassovi, and canicola. Agglutination tests performed with antigen prepared with one of the saprophytic biflexa isolates showed seropositive reactions in 99.1% of the equine sera, with agglutination titers ranging from 1:100 to 1:3,200. Absorption of selected horse sera with the saprophytic strain removed the agglutinins to Leptospira interrogans serotypes. This suggests the possibility that L. biflexa strains may act as an antigenic stimulus and account for some of the persistent multiple cross-reaction patterns of equine sera with pathogenic serotypes.
Significance of urinary isolates of coagulase-negative Micrococcaceae
Authors: Williams D N , Lund M E , Blazevic D J
Published: May 2021

Of 16,347 urine cultures submitted to the hospital laboratory, 68 (0.4%) specimens from 50 patients yielded greater than 10(4) coagulase-negative staphylococci/ml in pure culture. A total of 62 of 63 organisms available for study were staphylococci: 45 Staphylococcus epidermidis (predominantly subgroup 1), 15 Staphylococcus saprophyticus (subgroup 3), and 2 Staphylococcus aureus. Twenty-one patients had "probable" urine infections. Eight patients had two or more positive urine cultures, and all isolates from the same patients were identical (by morphology, antibiotic susceptibility, and hemolytic pattern). Nine (75%) of the 12 isolates of S. saprophyticus, which were novobiocin resistant and nonhemolytic on the synergistic hemolysis test, were from patients with probable urinary infection. Eight were young women with acute symptoms and pyuria. Differences in the glucose and mannitol fermentation tests with different media may lead to difficulties in identification. Novobiocin resistance cannot be relied upon to differentiate isolates of S. saprophyticus from S. epidermidis.
Microrespirometer chamber for determinations of viability in cell and organ cultures
Authors: Gabridge M G
Published: May 2021

The effects of chemical, physical, and infectious cytotoxic agents on primary and cultured cells were evaluated by measurements of oxygen uptake for various time periods. A newly developed respirometer used a Clark oxygen electrode in a 1.0-ml chamber, with provisions for constant mixing and for temperature control of both the sample and electrode chambers. The device was unique because the electrode and instrumentation were provided by a clinical blood-gas analyzer. Oxygen uptake by blank controls was negligible, whereas cells and tissue consumed oxygen at rates of approximately 1 to 5 mul/h in a dose- and temperature-dependent fashion. Cyanide, heat, and freeze-thaw lysis reduced the oxygen uptake to less than 0.6 mul/mg per h. Infection of trachea organ cultures with Mycoplasma pneumoniae significantly reduced relative ciliary activity, tetrazolium reduction capacity, and oxygen consumption in a coordinated fashion.
Combined serotyping and biotyping of Serratia marcescens
Authors: Rubin S J , Brock S , Chamberland M , Lyons R W
Published: May 2021

The API (Analytab Products, Inc., New York, N.Y.) biotypes of 117 clinical isolates of Serratia marcescens were determined and fell into 13 different patterns. The O and H antigens were determined by tube agglutination, and 27 serotypes were identified. The biotype and serotype appeared to vary indepently. Serotyping and biotyping combined divided these isolates into 56 different types. There was a problem interpreting the end points for inositol fermentation and urease production, which could affect reproducibility of API biotypes. Biotyping is a simple way of screening for possible nosocomial outbreaks of S. marcescens.
Serogrouping of Bacteroides fragilis subsp. fragilis by the agglutination test
Authors: Lambe D W , Moroz D A
Published: May 2021

The agglutination technique was used to establish a serological classification scheme for 98 strains of Bacteroides fragilis subsp. fragilis isolated from clinical specimens and normal human feces. Absorbed antisera were prepared to seven strains of B. fragilis subsp. fragilis. These seven absorbed antisera were species as well as subspecies specific and provided the basis of the serological classification scheme. This scheme was composed of 21 serogroups; seven of these serogroups contained only one group component. There was a total of 45 serological patterns. This serological scheme may be used for the serological classification of strains of B. fragilis subsp. fragilis and to study the epidemiology of this organism.
Systematic variables affecting simian virus 40-induced T-antigen expression and transformation in human cells
Authors: Kaplan M M , Lubiniecki A S , Blattner W A , Mason T , Giard D J , Gunnell M , Triantafellu N , Fraumeni J F
Published: May 2021

Simian virus 40 (SV40) infection of human skin fibroblast and human tumor cells resulted in the expression of T-antigen and transformed foci. By examining various conditions of input virus multiplicity and initial cell density, the systematic variation of T-antigen determination was minimized. The most uniform results were obtained at multiplicities of about 275 plaque-forming units/cell. Within limits (5 X 10(4) to 2 X 10(5) cells/dish), initial cell density had little effect on T-antigen expression. Volume of virus inoculum was critical for some cell lines, but not for others. Cell passage level had no general effect on T-antigen expression, although specific cell lines demonstrated increased or decreased levels of T-antigen expression with serial passage for no apparent reason. T-antigen expression correlated with virus-induced cell transformation (focus formation) at two different multiplicities. In addition, T-antigen assays at 3 days gave consistently more reproducible results than transformation assays at 21 days in seven cell lines tested at two multiplicities of infection. These results defined input multiplicity as the major source of systematic variability and will permit development of a more reproducible tool in the evaluation of individuals at high risk of cancer.
Counterimmunoelectrophoresis of staphylococcal antibody
Authors: Fedorko J , Fossieck B E , Parker R H
Published: May 2021

Modifications of the staphylococcal counterimmunoelectrophoresis technique were evaluated to determine how variations in the procedure affected results. Neither a buffer pH range of 7.8 to 9.0 nor buffer molarity of 0.015 or 0.025 when tested in combinations caused appreciable differences. However, use of different agar preparations or delay in addition of antigen to the test slide altered the location of the precipitin band. Agarose was found to be more sensitive in determining the serum precipitin titer and provided a better photographic record than either Ionagar or Noble agar.
Rapid micromethod of screening for antibodies to disease agents using the indirect enzyme-labeled antibody test
Authors: Saunders G C , Clinard E H
Published: May 2021

We have developed a rapid and simple indirect enzyme-labeled antibody procedure suitable for screening for antibodies to viral, bacterial, and parasitic antigens. Disposable microplates were utilized as antigen carriers and test vehicles. The test worked equally well with serum or whole unclotted blood. Using an automatic pipettor, 96 tests could be performed in 30 to 60 min. We report on the development of enzyme-labeled antibody procedures for the detection and surveillance of animal diseases.
Comparative isolation of vaginal yeasts on selective and nonselective media
Authors: Brashear M D , Wong J , Hines P , Smith R F
Published: May 2021

The isolation of vaginal yeasts was compared on a selective medium, phosphomolybdic acid agar, and on starch agar, a nonselective differential medium used primarily to isolate Corynebacterium vaginale. The majority of the Candida albicans strains were isolated on starch agar, but the selective medium was required for isolating all yeasts from the greatest number specimens.
Effect of volume of blood cultured on detection of bacteremia
Authors: Hall M M , Ilstrup D M , Washington J A
Published: May 2021

The rates of recovery of bacteria from vented vacuum blood culture bottles containing 50 and 100 ml of a soybean-casein digest broth were compared. Overall, more isolates were recovered from the larger bottle; moreover, gram-negative bacilli and especially Pseudomonas aeruginosa were recovered significantly more frequently (P less than 0.01) from the 100-ml bottle.
Development and Application of a PCR-Based Method Including an Internal Control for Diagnosis of Congenital Cytomegalovirus Infection
Authors: Jones Rachel N. , Neale M. Lynne , Beattie Brian , Westmoreland Diana , Fox Julie D.
Published: May 2021

Cytomegalovirus (CMV) is the most common cause of congenital infection in the developed world. We have designed and evaluated an assay that includes an internal control for amplification and detection of CMV DNA in amniotic fluid and neonatal urine samples. We present data on the use of this assay in the diagnosis of congenital CMV infection. A total of 145 amniotic and fetal fluid samples were examined by this assay; 83 were from healthy pregnant women and 62 were from women who were being investigated because of concerns over the pregnancy (diagnostic group). CMV DNA was detected in three amniotic fluid samples from the diagnostic group but was not detected in any samples taken from healthy pregnant women. Thirty-nine urine samples were obtained from 19 neonates with suspected congenital infection; CMV DNA was detected in urine from 6 of these patients. The assay provides useful information about CMV infection in the fetus and the neonate; when used in conjunction with other diagnostic tools it will enable mothers and obstetricians to make informed decisions about the management of pregnancies complicated by CMV infection.
Comparison of Antibody Titers Determined by Hemagglutination Inhibition and Enzyme Immunoassay for JC Virus and BK Virus
Authors: Hamilton R. S. , Gravell M. , Major E. O.
Published: May 2021

A comparison of antibody titers to JC virus (JCV) or BK virus (BKV) was made by hemagglutination inhibition (HI) and enzyme immunoassay (EIA) with 114 human plasma samples. Antibody titers to JCV or BKV determined by HI were lower than those determined by EIA. Nevertheless, as HI titers increased so did EIA titers. When antibody data were compared by the Spearman rank correlation test, highly significant correlations were found between HI and EIA titers. Results obtained by plotting EIA antibody titers for JCV against those for BKV generally showed a reciprocal relationship, i.e., samples with high antibody titers to JCV had lower antibody titers to BKV and vice versa. Some samples, however, had antibody titers to both viruses. Of the samples tested, 25.4% (25 of 114) had HI and EIA antibody titers to JCV and BKV which were identical or closely related. This is not the scenario one would expect for cross-reactive epitopes shared by the two viruses, but one suggesting that these samples were from individuals who had experienced infections by both viruses. Adsorption with concentrated JCV or BKV antigen of sera with high antibody titers to both JCV and BKV and testing by JCV and BKV EIA gave results which support this conclusion. Although 52.6% (51 of 97) of the samples from the Japanese population tested had very high antibody titers (≥40,960) to either JCV or BKV, none of the samples were found by a dot blot immunoassay to have antibodies which cross-reacted with simian virus 40. The results from this study, in agreement with those of others, suggest that humans infected by JCV or BKV produce antibodies to species-specific epitopes on their VP1 capsid protein, which is associated with hemagglutination and cellular binding.
In Vitro and In Vivo Evaluations of Sodium Lauryl Sulfate and Dextran Sulfate as Microbicides against Herpes Simplex and Human Immunodeficiency Viruses
Authors: Piret Jocelyne , Lamontagne Julie , Bestman-Smith Julie , Roy Sylvie , Gourde Pierrette , Désormeaux André , Omar Rabeea F. , Juhász Julianna , Bergeron Michel G.
Published: May 2021

The efficacy of sodium lauryl sulfate (SLS), a sulfated anionic chaotropic surfactant, and dextran sulfate (DS), a polysulfated carbohydrate, against herpes simplex virus (HSV) and human immunodeficiency virus (HIV) infections was evaluated in cultured cells and in different murine models of HSV infection. Results showed that both SLS and DS were potent inhibitors of the infectivities of various HSV-1 and HSV-2 strains. Pretreatment of HIV-1 (strain NL4-3) with SLS also reduced its infectivity to 1G5 cells. DS prevented the binding of HSV to cell surface receptors and therefore its entry into cells. Pretreatment of HSV-1 (strain F) with 50 μM SLS resulted in a complete loss of virus infectivity to Vero cells. However, viruses were able to enter into cells and to produce in the nuclei capsid shells devoid of a DNA core. The amount of the glycoprotein D gene produced in these cells remained unchanged compared to controls, suggesting that SLS could interfere with the maturation of the virus. At a higher SLS concentration (100 μM), HSV was highly damaged by SLS pretreatment and only a few viral particles could enter into cells to produce abnormal capsids. Although DS was a more potent inhibitor of HSV infectivity in vitro, it was unable to provide any protection in murine models of HSV infection. However, SLS conferred a complete protection of animals infected cutaneously with pretreated viruses. In addition, skin pretreatment of mice with a polymer formulation containing SLS completely prevented the development of cutaneous lesions. More interestingly, intravaginal pretreatment of mice with SLS in a buffered solution also completely protected against lethal HSV-2 infection. Taken together, our results suggest that SLS could thus represent a candidate of choice as a microbicide to prevent the sexual transmission of HIV, HSV, and possibly other pathogens that cause sexually transmitted diseases.
Isolation, Cultivation, and Characterization ofBorrelia burgdorferifrom Rodents and Ticks in the Charleston Area of South Carolina
Authors: Oliver J. H. , Clark K. L. , Chandler F. W. , Tao L. , James A. M. , Banks C. W. , Huey L. O. , Banks A. R. , Williams D. C. , Durden L. A.
Published: May 2021

Twenty-eightBorrelia burgdorferiisolates from the Charleston, S.C., area are described. This represents the first report and characterization of the Lyme disease spirochete from that state. The isolates were obtained from December 1994 through December 1995 from the tickIxodes scapularis, collected from vegetation, and from the rodentsPeromyscus gossypinus(cotton mouse),Neotoma floridana(eastern wood rat), andSigmodon hispidus(cotton rat). All isolates were screened immunologically by indirect immunofluorescence with monoclonal antibodies toB. burgdorferi-specific outer surface protein A (OspA) (antibodies H5332 and H3TS) andB. burgdorferi-specific OspB (antibodies H6831 and H614), aBorrelia(genus)-specific antiflagellin antibody (H9724),Borrelia hermsii-specific antibodies (H9826 and H4825), and two polyclonal antibodies (one toBorreliaspecies and another toB. burgdorferi). Six of the isolates were analyzed by exposing Western blots to monoclonal antibodies H5332, H3TS, H6831, and H9724. All isolates were also analyzed by PCR with five pairs of primers known to amplify selected DNA target sequences specifically reported to be present in the reference strain,B. burgdorferiB-31. The protein profiles of six of the isolates (two from ticks, one from a cotton mouse, two from wood rats, and one from a cotton rat) also were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We conclude that the 28 Charleston isolates areB. burgdorferisensu stricto based on their similarities to theB. burgdorferiB-31 reference strain.
The Efficacy of Laboratory Diagnosis of
Helicobacter pylori
Infections in Gastric Biopsy Specimens Is Related to Bacterial Density and
vacA
,
cagA
, and
iceA
Genotypes
Authors: van Doorn Leen-Jan , Henskens Yvonne , Nouhan Nathalie , Verschuuren Anita , Vreede Rolf , Herbink Paul , Ponjee Gabriëlle , van Krimpen Kees , Blankenburg Ruud , Scherpenisse Joost , Quint Wim
Published: May 2021

A total of 500 consecutive patients undergoing upper endoscopy were biopsied and tested for
H. pylori
infection by the
Campylobacter
-like organism (CLO) test, culture, histology, and PCR. Serum samples were tested by two different serological assays. Patients were considered
H. pylori
positive if at least two of the four biopsy specimen-based methods yielded positive results. PCR had the highest diagnostic sensitivity (99.4%), followed by histology (92.2%), culture (89.5%), and the CLO test (89.0%). The specificities of all methods were higher than 98%. Of the organisms from the 181 PCR-positive patients, the
vacA
(s and m regions),
cagA
, and
iceA
genotypes were determined by reverse hybridization (line probe assay) or an allele-specific PCR. Organisms that were detected by PCR but that remained undetected by the CLO test were significantly more often
vacA
s1 (
P
= 0.006), m1 (
P
= 0.028), and
cagA
positive (
P
= 0.029) than
vacA
s2, m2, and
cagA
negative, respectively. Organisms that were detected by PCR but that remained undetected by culture or histology more often contained
iceA1
(
P
= 0.034 and
P
= 0.029, respectively) than
iceA2
. Higher
H. pylori
density was associated with
vacA
s2 (
P
= 0.024),
vacA
m2 (
P
= 0.050), and
cagA
-negative (
P
= 0.035) genotypes. Also, the diagnostic results of the CLO test (
P
= 0.001) and culture (
P
= 0.031) but not those of the PCR (
P
= 0.130) were significantly associated with the
H. pylori
density. The rate of detection by the four biopsy specimen-based tests was lower for patients who used proton pump inhibitors, but this was independent of the
H. pylori
genotypes. These observations may be explained by different bacterial densities, as established by the distinct genotypes of
H. pylori
, and confirm that the biologies of strains with such genotypes are considerably different.
Specific
Taenia crassiceps
and
Taenia solium
Antigenic Peptides for Neurocysticercosis Immunodiagnosis Using Serum Samples
Authors: Bueno Ednéia Casagranda , Vaz Adelaide José , Machado Luís Dos Ramos , Livramento JoséAntônio , Mielle Sílvia Regina
Published: May 2021

Neurocysticercosis (NC), i.e., the presence of the larval form of
Taenia solium
in tissues, is the most frequent and severe infection involving the central nervous system. Paired serum and cerebrospinal fluid (CSF) samples from patients with NC, CSF and serum samples from a control group, and serum samples from patients with other parasitoses were studied by enzyme-linked immunosorbent assay (ELISA) and by immunoblotting with
Taenia crassiceps
vesicular fluid antigen (Tcra) and
Taenia solium
total saline antigen (Tso) for the detection of immunoglobulin G antibodies. ELISAs carried out with the Tso and Tcra antigens showed 94.1 and 95.6% sensitivities, respectively, for the detection of antibodies in CSF and 70.6% and 91.2% sensitivities, respectively, for the detection of antibodies in serum, with 100% specificity for the detection of antibodies in CSF and 80% specificity for the detection of antibodies in serum for both antigens. On the basis of the reactivities of the peptides in the samples analyzed, the peptides of ≤23, 39, 85 to 77, and 97 kDa were found to be Tso specific by immunoblotting and the peptides of ≤62, 74, 109, 121, and 131 kDa were found to be Tcra specific. Tests with Tcra extract had higher sensitivities and more homogeneous results and permitted us to obtain the parasites easily. We suggest the use of Tcra ELISA for the study of serum and confirmation of the results for sera positive by an immunoblotting analysis in which specific peptides (e.g., peptides of 19 to 13 kDa) are detected.
Comparison of PCR-Restriction Fragment Length Polymorphism Analysis and PCR-Direct Sequencing Methods for Differentiating
Helicobacter pylori ureB
Gene Variants
Authors: Tanahashi Toshihito , Kita Masakazu , Kodama Tadashi , Sawai Naoki , Yamaoka Yoshio , Mitsufuji Shoji , Katoh Fumitaka , Imanishi Jiro
Published: May 2021

A method utilizing PCR-restriction fragment length polymorphism (RFLP) in the
Helicobacter pylori
genes is widely used to differentiate strains. However, with this typing method only a single base change at a specific restriction site can be detected. In addition, it is unclear whether the nucleotide base change recognized by RFLP is related to a substitution of encoded amino acid. To examine the validity of the PCR-RFLP method, 933-bp PCR products were obtained from 41 different clinical
H. pylori
isolates and were digested with
Sau
3A restriction endonuclease. Furthermore, the nucleotides of the same region in the
ureB
gene were directly sequenced and compared. PCR-RFLP confirmed that there was genetic diversity within the
ureB
gene with three distinct types, one being well conserved and the other two being variations. However, the direct sequencing method revealed that there was no difference at the nucleotide level among these RFLP types. Base substitutions recognized by
Sau
3A occurred in the third-base position and did not change the encoded amino acid. In addition, many nucleotide mutations, which could not be recognized by
Sau
3A, were frequently found. These results suggest that the PCR-RFLP method provides for an easy typing scheme of isolates, but does not reveal the true extent of genetic diversity. It is proposed that careful observation is required for the interpretation of results when clinical isolates are differentiated.
Serodiagnosis of Recently Acquired
Toxoplasma gondii
Infection with a Recombinant Antigen
Authors: Li Shuli , Maine Greg , Suzuki Yasuhiro , Araujo Fausto G. , Galvan Gina , Remington Jack S. , Parmley Stephen
Published: May 2021

A portion of a cDNA encoding a 35-kDa antigen from
Toxoplasma gondii
was cloned into the CKS expression vector and expressed in
Escherichia coli
. By using the enzyme-linked immunosorbent assay (ELISA), the recombinant protein (rP35 antigen) was examined for reactivity with immunoglobulin G (IgG) antibodies in the sera of pregnant women. Of these women, 41 had a toxoplasma serologic profile suggestive of recently acquired
T. gondii
infection (Sabin-Feldman dye test [DT] titers from 1:256 to 1:32,000, positive IgM ELISA titers from 2.3 to 9.7, positive IgA ELISA from 1 to >28, and acute patterns in the differential agglutination [AC/HS] test) (group I), and 50 women had a toxoplasma serologic profile suggestive of infection acquired in the distant past (low DT titers from 1:16 to 1:512, negative IgM ELISA titers from 0 to 0.8, and chronic patterns in the AC/HS test) (group II). The classification of acute or chronic profile was based on the individual's clinical history as well as the combination of the results of the toxoplasma serological profile. An additional group (group III) was composed of sera from 50 women who were seronegative for
T. gondii
antibodies in the DT. The results revealed that whereas 85.3% of women in group I had IgG antibodies that reacted with the rP35 antigen, only 8% of women in group II had IgG antibodies that reacted with the same antigen. In immunoblots, the rP35 antigen was recognized by IgG antibodies in a pool of sera from individuals with a toxoplasma serologic profile compatible with acute infection but not in a pool of sera from individuals with a serologic profile characteristic of a chronic infection. These results reveal that IgG antibodies against the P35 antigen are produced during the acute stage of the infection but are uncommon in the latent or chronic phase of the infection. Thus, the rP35 antigen may be a useful serologic marker to differentiate between recently acquired infection and that acquired in the more distant past.