bluuebunny/crossref_metadata_2025 huggingface dataset:
Laboratory diagnosis of gonococcal infection by genetic transformation
Authors: Butler L O , Knight R D
Published: January 2020

The transformation test for the detection of infection by Neisseria gonorrhoeae has been examined using pro gonococci as recipients and DNA preparations from 912 clinical isolates and from 240 direct swab specimens as donors. The reliability of the method was checked with DNA from clinical isolates; 82% of the N. meningitidis from throat swab specimens were capable of transforming the gonococcal recipients, but after identification of the meningococcus by the aminopeptidase profile, the transformation test was then 99.5% positive for the gonococcus with virtually no false-positives. The only other organism to give a positive reaction was N. lactamica, which occurred once in 912 specimens. When applied directly to swab specimens, the reliability of the test was reduced, but this may have been related to variability of the specimen itself. However, 7 of 15 specimens which were microscopically suspected to be gonococci but unculturable were positive; also, 9 out of 38 unculturable specimens that were not even suspected to be gonococci were positive. Hence the test was able to identify the presence of gonococci that were unculturable. The aminopeptidase activities were not sensitive enough to be detected in the direct swab specimens, and neither cys nor leu auxotrophs were suitable as recipients to give a differentiation between N. gonorrhoeae and N. meningitidis. Evidence was obtained which would support the proposition that the transfer of genetic material between N. gonorrhoeae and N. meningitidis may occur.
Development of a simple serological method for diagnosing leptospirosis: a microcapsule agglutination test
Authors: Arimitsu Y , Kobayashi S , Akama K , Matuhasi T
Published: January 2020

A passive microcapsule agglutination test for the diagnosis of leptospirosis was developed by utilizing chemically stable microcapsules instead of sheep erythrocytes. In the test, sonically disrupted antigens of leptospira were sensitized to microcapsules treated with glutaraldehyde. Compared with the microscopic agglutination test, the passive microcapsule agglutination test showed a relatively genus-specific tendency and a 4- to 32-fold-higher sensitivity. The sensitized microcapsule antigens were stable for at least 1 year. The microcapsules coupled with mixed antigens can be used as a serodiagnostic screening test for diseases caused by various types of leptospira. The test, which is very simple and reproducible and requiring no specific training, can be employed easily as a routine test in diagnostic laboratories.
Iontophoretic killing of Escherichia coli in static fluid and in a model catheter system
Authors: Davis C P , Arnett D , Warren M M
Published: January 2020

A static iontophoretic model system was used to determine the effects of various metals, microamperage (microamperemeter) levels, and voltage levels on Escherichia coli and several other bacterial genera. Only gold iontophoresis at 400 microamperemeter and approximately 3.2 V was found to kill all bacterial genera tested when the inoculum dose was 1 x 10(3) cells per ml. E. coli was killed at all inocula doses to 1 x 10(7), whereas other genera were inhibited. Other metals, such as silver or copper, were not effective in killing bacteria. A model catheter system, which contained a gold iontophoresis device, was used to kill E. coli in a flowing system at sites as far as 10 cm downstream from the iontophoretic device. The experiments suggest gold iontophoresis can be a potential tool in controlling catheter-related infections.
Increased Efficiency of Group B Coxsackievirus Isolation from Clinical Specimens by Use of BGM Cells
Authors: Menegus Marilyn A. , Hollick Gary E.
Published: January 2020

A continuous African green monkey kidney cell line, designated BGM, was compared with primary cynomolgus monkey kidney cells and human embryonic lung cells for efficiency of enterovirus isolation. A selective enhanced sensitivity of BGM cells both in terms of isolation rate and speed of isolation was found for group B coxsackieviruses but could not be demonstrated for a number of other nonpolio enteroviruses.
In vitro evaluation of CENTA, a new beta-lactamase-susceptible chromogenic cephalosporin reagent
Authors: Jones R N , Wilson H W , Novick W J , Barry A L , Thornsberry C
Published: January 2020

CENTA is a newly synthesized, beta-lactamase-labile, chromogenic cephalosporin reagent which changes color from light yellow (lambda maximum ca. 340 nm) to chrome yellow (lambda maximum ca. 405 nm) concomitant with hydrolysis of the beta-lactam ring. This compound offers promise as a diagnostic reagent comparable to other chromogens (PADAC and nitrocefin) for the early detection of beta-lactamase-producing clinical isolates, while retaining some antimicrobial effect against Escherichia coli, Klebsiella spp., Proteus mirabilis, Staphylococcus aureus, and non-enterococcal Streptococcus spp. CENTA is relatively unaffected by commonly used microbiological media and human serum.
Predominant microflora associated with human dental periapical abscesses
Authors: Oguntebi B , Slee A M , Tanzer J M , Langeland K
Published: January 2020

The microflora of periapical abscesses of teeth free of periodontal disease was studied. Permucosal aspiration of abscess contents and anaerobic cultural, microscopic, and biochemical techniques indicated a mixed but somewhat specific and relatively limited facultative and obligate anaerobic flora with Fusobacterium nucleatum and Streptococcus mitis as a frequent pair.
Comparison of direct and indirect immunofluorescence staining of clinical specimens for detection of respiratory syncytial virus antigen
Authors: Minnich L L , Ray C G
Published: January 2020

Immunofluorescence staining methods for respiratory syncytial virus antigen detection were compared. Of 50 specimens originally positive for respiratory syncytial virus by direct immunofluorescence and culture, 49 were positive by repeat direct immunofluorescence and 32 were positive by indirect immunofluorescence. Additional results obtained on specimens originally negative for respiratory syncytial virus by direct immunofluorescence, culture, or both indicate that direct immunofluorescence staining for respiratory syncytial virus antigen was more sensitive than was indirect immunofluorescence.
Rapid biotyping of Haemophilus influenzae and Haemophilus parainfluenzae with PathoTec strips and spot biochemical tests
Authors: Juni B A , Rysavy J M , Blazevic D J
Published: January 2020

PathoTec strips and spot biochemical tests were evaluated for the ability to biotype Haemophilus influenzae and Haemophilus parainfluenzae. Indole, urease, and ornithine decarboxylase reactions were tested. The results of PathoTec strips compared favorably with those conventional methods; the percent agreements were as follows: indole, 100; urease, 99.5; and ornithine, 95.5. Spot tests were simple and rapid, and the results also compared favorably with those of conventional tests; the percent agreements were as follows: indole, 99; urease, 100; and ornithine, 96.
Serological diversity in Haemophilus somnus
Authors: Canto G J , Biberstein E L
Published: January 2020

Serological interrelations among 46 strains of Haemophilus somnus of various geographical and pathological origins were studied by agglutination and agglutinin adsorption procedures. Sera prepared against nine representative American and Swiss isolates agglutinated all other strains to various titers. Adsorption of antisera to American strains by Swiss cultures tended to remove mainly the reactivity with Swiss antigens, whereas adsorption of such antisera to American strains in most instances abolished all agglutinating activity. Analogous observations were made when sera against Swiss cultures were adsorbed with cells of American and Swiss origin, respectively. Results of tests involving unadsorbed and adsorbed sera suggest the existence of at least three sets of antigen in H. somnus: American, Swiss, and common, which appear to exist in various combinations, accounting for four agglutination groups. These reflect to a considerable degree the geographical, although not the pathological or anatomical, origin of the respective strains.
Growth of Nosocomial Pathogens in Multiple-Dose Parenteral Medication Vials
Authors: Highsmith Anita K. , Greenhood Gary P. , Allen James R.
Published: January 2020

The extent to which microbial contamination of medications dispensed in multiple-dose vials might serve as a source of infection to patients has not been fully investigated. To characterize the effects of microbial contamination, we studied the growth-supporting properties of eight medications dispensed in multiple-dose vials. Two medications, procainamide and methohexital, demonstrated no survival of any microbes 24 h after inoculation. Succinylcholine chloride, regular insulin, potassium chloride, heparin, and thiopental slowly killed or allowed limited survival of several of the microorganisms used as contaminants. Lidocaine allowed survival or proliferation of several microbial strains suspended in 0.25% peptone water in saline, but slowly killed all strains except
Pseudomonas cepacia
suspended in 0.9% saline. Endotoxin, measured by the
Limulus
amebocyte lysate assay, was found in the two medications tested, lidocaine contaminated with
Pseudomonas cepacia
and insulin contaminated with enterococci. Inadvertent microbial contamination of at least some parenteral medications in multiple-dose vials may result in the exposure of patients to viable organisms. The potential, however, for medications such as lidocaine to support growth of organisms under selected circumstances should be noted by those responsible for preparing and administering these drugs. The potential hazard to patients from endotoxin in contaminated medications under these circumstances has not been assessed. Additional studies of this type should be pursued to provide more complete information about the risk of microbial contamination of products for parenteral use.
Detection of specific antibody by enzyme-linked immunosorbent assay and antigenemia by counterimmunoelectrophoresis in humans infected with Pneumocystis carinii
Authors: Maddison S E , Hayes G V , Slemenda S B , Norman L G , Ivey M H
Published: January 2020

A urea-soluble extract of cyst-rich material from rat lung heavily infected with Pneumocystis carinii was evaluated in an enzyme-linked immunosorption assay for antibody in 461 human sera. The highest level of reactivity occurred in sera submitted for serodiagnosis from proved or highly suspect cases. However, the range of reactivities in these groups, many of whom were on immunosuppressive therapy, was very wide. A more restricted lower range of reactivity was observed in both hospital-family contacts and healthy Serum Bank donors. Because of the overlap in levels of reactivity between the pneumocystosis and control groups, no concise cutoff value to separate infected from noninfected individuals could be made. Specificity of the reactions was shown by absorption of patients' and control sera with uninfected and P. carinii-infected human and rat lung tissue. The data support the concept that P. carinii is highly prevalent as a latent agent in the general population and is provoked to cause clinically manifest disease in the compromised host. Detection of circulating antigen appeared to be specific and possibly a useful adjunct to diagnosis, as 10 of the 14 proved or highly suspect patients with antigenemia did not have measurable antibody to P. carinii.
Isopycnic separation of Escherichia coli cultures possessing colonization factor antigen I
Authors: Giesa F R , Zajac I , Bartus H F , Actor P
Published: January 2020

A culture of Escherichia coli possessing colonization factor antigen I was subjected to isopycnic separation on Percoll gradients. The results demonstrated successful division of the culture into two populations: (i) bacteria which cause mannose-resistant hemagglutination and (ii) bacteria which lack the ability to hemagglutinate in the presence of mannose.
Isolation of Legionella pneumophila from hospital potable water specimens: comparison of direct plating with guinea pig inoculation
Authors: Edelstein P H , Snitzer J B , Finegold S M
Published: January 2020

Eighty-five potable water samples were analyzed for the presence of Legionella pneumophila by direct plating and by guinea pig inoculation methods. Direct plating methods utilized antimicrobial-containing media with and without acid treatment of specimens before inoculation. Thirty-three specimens were culture positive for L. pneumophila by one or both techniques. A total of 14 specimens were positive for L. pneumophila by direct plating only, and 2 specimens were positive for L. pneumophila by guinea pig inoculation only. The sensitivity of direct plating is significantly greater than that of guinea pig inoculation when isolating L. pneumophila from potable water (P less than 0.01).
Identification of Proteus penneri sp. nov., formerly known as Proteus vulgaris indole negative or as Proteus vulgaris biogroup 1
Authors: Hickman F W , Steigerwalt A G , Farmer J J , Brenner D J
Published: January 2020

The name Proteus penneri sp. nov. is proposed for a group of organisms previously called Proteus vulgaris indole negative or P. vulgaris biogroup 1. All of these strains were salicin negative, esculin negative, and chloramphenicol resistant (zone size, less than 14 mm). DNA relatedness studies indicated that when DNA from P. penneri strain 1808-73 was labeled and tested against unlabeled DNA from 13 other P penneri strains, a highly related group was formed (88 to 99% relatedness at 60 degrees C and 67 to 99% relatedness at 75 degrees C). Strain 1808-73 (ATCC 33519) is proposed as the type strain of P. penneri. In this study, two distinct groups of indole-positive P. vulgaris strains were also apparent. The first group (defined as P. vulgaris biogroup 2) was indole positive, salicin positive, and esculin positive, and the second group (defined as P. vulgaris biogroup 3) was indole positive, salicin negative, and esculin negative. The current type strain of P. vulgaris (ATCC 13315) belongs to biogroup 3. The DNA from P. penneri strains was not highly related to labeled DNA from the type strain of P. vulgaris (14 to 30% relatedness at 75 degrees C) or from P. vulgaris strain PR 1 (ATCC 29905), which belongs to biogroup 2 (27 to 33% relatedness at 75 degrees C). Strains of biogroup 2 were sensitive to chloramphenicol (zone size, greater than 19mm), and 10 of these strains formed a highly related group by DNA hybridization when DNA from PR 1 was labeled (64 to 100% relatedness at 60 degrees C and 70 to 100% relatedness at 75 degrees C), but they were not highly relatedness to the type strain of P. vulgaris (51 to 68% relatedness at 60 degrees C and 14 to 44% relatedness at 75 degrees C). Further DNA relatedness studies are needed on strains of biogroup 3 before a definitive taxonomic proposal can be made for these two indole-positive biogroups.
Serodiagnosis of tsutsugamushi fever (scrub typhus) by the indirect immunoperoxidase technique
Authors: Yamamoto S , Minamishima Y
Published: January 2020

The indirect immunoperoxidase technique was assessed for the serodiagnosis of tsutsugamushi fever (scrub typhus). The antigens were peritoneal smears prepared from mice infected intraperitoneally with the Karp, Kato, and Gilliam strains of Rickettsia tsutsugamushi. Treatment of the mice with cyclophosphamide apparently increased the number of the rickettsiae, and it minimized the exudate that interfered with the specific staining. The rickettsiae were seen as clusters in the juxtanuclear region of the mesothelial cells and also as free particles outside of the cells. By the indirect immunoperoxidase technique, the sera from all of the patients (49 samples from 30 patients) were positive for the R. tsutsugamushi antibody. The antibody titers (immunoglobulin G [IgG] and IgM) determined by the indirect immunoperoxidase technique correlated with those determined by the indirect immunofluorescence technique. Thus, the indirect immunoperoxidase technique was useful for quantifying both IgG and IgM antibodies to the rickettsia.
Pseudooutbreak of cryptococcal meningitis
Authors: Hopfer R L , Katz R L , Fainstein V
Published: January 2020

A pseudooutbreak of cryptococcal meningitis was caused by the use of contaminated albumin solution used in the preparation of Papanicolaou-stained slides of cerebrospinal fluid. Organisms were seen in cytocentrifuge preparations, but not in India ink preparations of cerebrospinal fluid specimens. Cryptococcal antigen tests were positive and Cryptococcus neoformans was cultured from the albumin-treated cerebrospinal fluid specimens and from the albumin solution.
Growth of non-Campylobacter, oxidase-positive bacteria on selective Campylobacter agar
Authors: Moskowitz L B , Chester B
Published: January 2020

A total of 67 oxidase-positive, gram-negative bacteria were tested for growth on selective Campylobacter agar (Blaser formulation, BBL Microbiology Systems, Cockeysville, Md.) at 42 degrees C under microaerophilic conditions. Although the growth of most of these bacteria was prevented, all strains of Achromobacter xylosoxidans, Pseudomonas aeruginosa, Pseudomonas putrefaciens, Pseudomonas alcaligenes, and Pseudomonas pseudoalcaligenes grew as well as Campylobacter fetus subsp. jejuni.
New virulence-associated plasmid in Yersinia enterocolitica
Authors: Kay B A , Wachsmuth K , Gemski P
Published: January 2020

In a study of 103 strains of Yersinia enterocolitica, 10 strains were found to be lethal for mice and to possess 42- and 82-megadalton plasmids. This association was statistically significant (P much less than 0.001). Serotypes of Y. enterocolitica previously considered avirulent were found to possess these plasmids and to be lethal for mice. A spontaneous derivative of one strain contained only the 82-megadalton plasmid and was lethal for mice anyway. This virulence-associated plasmid is a potential diagnostic tool for the clinical or public health laboratory which must delineate pathogenic strains of Y. enterocolitica.
Preliminary identification of beta-hemolytic streptococci in throat swab cultures with a commercial blood agar slide (streptocult)
Authors: Christensen P , Danielsson D , Hovelius B , Kjellander J
Published: January 2020

A commercial blood agar slide (Streptocult, Orion Diagnostica) was used for preliminary identification of beta-hemolytic streptococci of groups A, C, and G in throat swab specimens. The sensitivity of the test was 93.6% and the specificity was 94.7%, as judged from 580 specimens. A model is suggested for routine processing of throat swab specimens, involving inoculation and reading of the slide in general practice and transport of positive or inconclusive slides to a bacteriology laboratory for isolation and serological grouping of beta-hemolytic streptococci. The model combines preliminary detection of beta-hemolytic streptococci within 24 h with the reliability of serological groupings, and should reduce the volume of specimens sent to the laboratory considerably.
Enzyme-linked immunosorbent assay for measurement of antibody to type III group B streptococci
Authors: Polin R A , Douglas S D , Kasper D L , Baker C J
Published: January 2020

Neonates at risk for fulminant type III, group B streptococcal (III GBS) infection are those who lack antibody to the capsular polysaccharide. A newly developed enzyme-linked immunosorbent assay (ELISA) was compared with a standard radioactive antigen-binding assay (RABA) for quantitation of III GBS antibody in human sera. Although there was a significant correlation between the ELISA and RABA (r = 0.81; P less than 0.001) in general, the ELISA detected antibody both to core and native antigens of III GBS, whereas the RABA detected antibody to native polysaccharide exclusively. The results of the two assays were discordant when sera which had only high native or core antibody (not both) were assessed. Although the ELISA was reproducible and required less than 1 microliter of serum, interpretation of data obtained by the assay should be viewed with caution since only antibody to native III GBS has been correlated with human immunity.
Isolation of Treponema hyodysenteriae from wild rodents
Authors: Joens L A , Kinyon J M
Published: January 2020

Rodents from swine-producing farms were examined for the presence of Treponema hyodysenteriae. Wild mice (n = 257) and rats (n = 41) were trapped on eight farms. Ceca were removed aseptically, and the contents and mucosal scrapings were cultured on selective medium (blood agar containing 400 micrograms of spectinomycin per ml). T. hyodysenteriae was detected in the cecal scrapings of four mice from three different farms where swine dysentery had occurred. Gross lesions were detected in the ceca in two of the four mice. In addition, Treponema innocens was detected in the cecal scrapings of 12 mice and 13 rats. Three of the four T. hyodysenteriae isolates were pathogenic when inoculated intragastrically into swine. The results of this investigation suggest that wild rodents may be carriers of T. hyodysenteriae.
Detection of Legionella antigenuria by reverse passive agglutination
Authors: Tang P W , de Savigny D , Toma S
Published: January 2020

A reverse passive agglutination method was developed to detect soluble antigens of Legionella spp. By this method Legionella antigens were detected in urine specimens from 14 of 15 antigenuric patients with clinically diagnosed Legionnaires disease and in none of 263 urine samples from healthy subjects or patients with urinary tract infections. Intra-genus cross-reactivity was observed only between L. pneumophila serogroups 2, 3, and 6. The Legionella reverse passive agglutination method was also evaluated with reference to reagent concentrations, test conditions, and subjectivity of reading test results. The method is rapid and does not require special equipment.
Comparative evaluation of supplemented peptone broth with sodium polyanetholesulfonate and trypticase soy broth with sodium amylosulfate for detection of septicemia
Authors: Tenney J H , Reller L B , Wang W L , Cox R L , Mirrett S
Published: January 2020

We compared the yield and speed of detection of clinically important microorganisms from 10,156 paired 5-ml samples of blood cultured in supplemented peptone broth (SPB) with 0.03% sodium polyanetholesulfonate (SPS) or Trypticase soy broth (TSB) with 0.5% sodium amylosulfate (SAS). The atmosphere of incubation (open venting units) and ratio of blood to broth (1:10) were the same for both samples. Only cultures with adequate blood samples (greater than or equal to 80% of stated volume) were compared statistically. Overall, SPB/SPS outperformed TSB/SAS. Bacteroidaceae and Eubacterium were found more often (P less than 0.05) and viridans streptococci were found sooner (P less than 10(-4)) in SPB/SPS than in TSB/SAS. Most importantly, staphylococci were found both more often (P less than 0.03) and sooner (P less than 10(-7)) in SPB/SPS than in TSB/SAS. In a separate experiment, SAS slowed the growth of a clinical strain of Staphylococcus aureus in TSB. Unless important advantages can be confirmed for SAS in controlled clinical trials, SAS cannot be recommended for routine use as an anticoagulant in blood culture media.
Whole-bacterial cell enzyme-linked immunosorbent assay for Streptococcus sanguis fimbrial antigens
Authors: Elder B L , Boraker D K , Fives-Taylor P M
Published: January 2020

A whole-bacterial cell enzyme-linked immunosorbent assay (bactELISA) was developed for detecting fimbrial antigens on Streptococcus sanguis. In this assay, S. sanguis cells were directly adhered to polystyrene or polyvinyl via drying. Use of the assay indicated that consistently high and uniform optical densities could be obtained from well to well. In addition, radioactive assaying indicated increased adsorption to the polystyrene wells over polyvinyl, suggesting that polystyrene may prove superior in the gram-positive bactELISA. Use of the bactELISA may prove valuable to both the clinical and research laboratory involved in the study of bacterial cell surface components or in the evaluation of antisera directed against bacterial antigens, which are difficult to prepare as purified derivatives.
Gluteal abscess caused by Phialophora hoffmannii and review of the role of this organism in human mycoses
Authors: Rinaldi M G , McCoy E L , Winn D F
Published: January 2020

Infections caused by members of the Phialophora hoffmannii (Beyma) Schol-Schwarz fungal aggregate (a group of related taxa, especially species of uncertain circumscription) are not reported frequently. This case report concerns the development of a gluteal abscess after multiple intramuscular injections of antimicrobial agents. Microbiological examination of the abscess material yielded a pure growth of the mold P. hoffmannii. Hyphal elements in the purulent exudate were only demonstrated when the specimen was digested by an N-acetyl cysteine-NaOH digestion-decontamination procedure or in stained paraffin block sections of the exudate. Other cases of disease caused by members of this fungal aggregate and the status of the mycology of this organism are reviewed.
New Salmonella serotype: Salmonella enteritidis serotype Grandhaven (30(1):r:1,2)
Authors: McDougal D L , Treleaven B E , Renshaw E C
Published: January 2020

A new Salmonella serotype, Salmonella enteritidis serotype Grandhaven (30(1):r:1,2), was isolated from the stool of a 35-year-old man with mild gastroenteritis. He had just returned from Sudan, Africa.
Detection of human Fc (gamma) receptors on streptococci by indirect immunofluorescence staining: a survey of streptococci freshly isolated from patients
Authors: Lebrun L , Pillot J , Grangeot-Keros L , Rannou M T
Published: January 2020

With immunofluorescence staining, Fc (gamma) receptors were found on groups A, C, and G streptococci and never on pneumococci or on groups B, D, H, or K streptococci.
Detection by radioimmunoassay and enzyme-linked immunosorbent assay of coronavirus antibodies in bovine serum and lacteal secretions
Authors: Rodak L , Babiuk L A , Acres S D
Published: January 2020

The sensitivity of a radioimmunoassay (RIA), an enzyme-linked immunosorbent assay (ELISA), and a serum neutralization assay (SN) for detecting antibodies to bovine coronavirus in serum and colostrum were compared. Although there proved to be a good correlation among all three assays (r = 0.915 and 0.964 for RIA with SN and ELISA, respectively), RIA and ELISA proved to be at least 10 times more sensitive than neutralization tests. By using these techniques, it was possible to detect a time-dependent decrease in antibody levels in bovine colostrum after parturition. Using ELISA, we demonstrated that 12 of 12 herds in Saskatchewan, and 109 of 110 animals tested, and antibody to bovine coronavirus. There was no elevated antibody response in serum or lacteal secretions of cows vaccinated once or twice with a commercially available modified live rota-coronavirus vaccine. In addition to being more sensitive than SN, ELISA and RIA proved to have other advantages for measuring antibody levels to bovine coronavirus and therefore warrant wider use as tools in diagnostic virology.
Sensitivity of immunofluorescence with monoclonal antibodies for detection of Chlamydia trachomatis inclusions in cell culture
Authors: Stephens R S , Kuo C C , Tam M R
Published: January 2020

Monoclonal antibodies which recognize the species-specific major outer membrane protein antigen of Chlamydia trachomatis were used for immunofluorescence staining of chlamydial inclusions in cell culture. A total of 115 clinical specimens were inoculated onto replicate HeLa 229 cell monolayers and assayed for chlamydial inclusions by immunofluorescence staining and Giemsa staining. Of the isolates, 38 were detected by immunofluorescence staining on passage 1 and 1 was detected on passage 2; 23 isolates on passage 1 and 13 isolates on passage 2 were detected by Giemsa staining. Immunofluorescence staining was significantly more sensitive than Giemsa staining for detecting chlamydial inclusions, particularly from specimens containing low titers of Chlamydia.
Diagnostic value of the enzyme-linked immunosorbent assay for detection of Candida albicans cytoplasmic antigen in sera of cancer patients
Authors: Araj G F , Hopfer R L , Chesnut S , Fainstein V , Bodey G P
Published: January 2020

To diagnose systemic candidiasis we developed an enzyme-linked immunosorbent assay (ELISA) for the quantitation of Candida albicans cytoplasmic antigen in sera of cancer patients. The antigen concentration was evaluated in 59 patients divided into several groups according to clinical, histological, and microbiological findings as well as antimicrobial treatment. All of seven patients with proven disseminated candidiasis had elevated antigen concentrations (mean = 1,200 ng/ml). Among 13 patients with suspected disseminated candidiasis, 9 had elevated antigen concentrations (mean = 1,039 ng/ml). The remaining four patients showed negative antigen (less than 125 ng/ml). In a group of patients with fever of unknown origin, 5 of 17 had high antigen concentrations (mean = 1,220 ng/ml) and the rest were negative. Patients with bacterial infection (6 patients), aspergillosis (3), cryptococcosis (4), coccidioidomycosis (3), and no infection (6), as well as normal healthy adults (31), had antigen concentrations of less than 100 ng/ml. This ELISA is rapid, sensitive, and specific and is a helpful diagnostic tool in the evaluation of candidiasis in cancer patients.
Further evaluation of the Biken test (modified Elek test) for detection of enterotoxigenic Escherichia coli producing heat-labile enterotoxin and application of the test to sampling of heat-stable enterotoxin
Authors: Honda T , Arita M , Takeda Y , Miwatani T
Published: January 2020

A total of 2,229 strains of Escherichia coli isolated from subjects with traveller's diarrhea were tested for heat-labile enterotoxin production by the Biken test (Honda et al., J. Clin. Microbiol. 13:1--5, 1981). Results of the Biken test showed a 99.0% coincidence with those obtained by Chinese hamster ovary cell assay. The use of antiserum against E. coli heat-labile enterotoxin gave sharper and clearer results than did the use of antiserum against cholera enterotoxin. Samples for heat-stable enterotoxin assay were obtained from the agar plates used for the Biken test, and the heat-stable enterotoxin activities of these samples were compared with those of samples obtained from standard liquid cultures. Results for samples of 2,229 strains from the Biken agar plates and from liquid cultures were almost identical.
Measurement of immunoglobulin G and M antibodies to type 3 pneumococcal capsular polysaccharide by enzyme-linked immunosorbent assay
Authors: Carlson B A , Giebink G S , Spika J S , Gray E D
Published: January 2020

An enzyme-linked immunosorbent assay was developed to measure immunoglobulin G (IgG) and IgM type 3 antipneumococcal capsular polysaccharide antibodies. The use of Fab2 fragments of rabbit antipneumococcal IgG antibody in the antibody-antigen sandwich increased the sensitivity for measuring IgM antibodies and decreased background activity in antigen-free cuvettes. This methodology detected type 3 IgM antibody responses in six of six subjects vaccinated with polyvalent pneumococcal vaccine and detected type 3 IgG antibody responses in three subjects. Results of the enzyme-linked immunosorbent assay and radioimmunoassay procedures were concordant, and postvaccination enzyme-linked immunosorbent assay IgM titers showed a stronger correlation with total radioimmunoassay antibody than did postvaccination ELISA IgG titers.
Evaluation of the MS-2 urine screening method for detection of bacteriuria
Authors: McCarthy L R , Gavan T L , Robson J , Corlett C
Published: January 2020

A collaborative evaluation of the MS-2 urine screening system indicated that the automated method could successfully identify at least 84% of urine specimens that contained greater than or equal to 10(5) colony-forming units per ml. Most specimens not detected as positive by the system contained primarily diphtheroids and yeasts. Excluding those specimens which contained diphtheroids and yeasts, the MS-2 system correctly identified as positive 94.8% of specimens that contained greater than or equal to 10(5) colony-forming units per ml and as negative 96.1% of specimens that contained less than 10(4) colony-forming units per ml. The automated system is an attractive method for the screening of urine specimens for significant bacteriuria.
Comparison of a fluorometric method with radial immunodiffusion assays for determination of complement components C3 and C4
Authors: Koelle M , Bartholomew W R
Published: January 2020

Measurements of patient serum complement components C3 and C4 are useful indicators of complement consumption in immune complex diseases. A fluorometric quantitative immunofluorescence system was evaluated in terms of measuring these complement components, and the results were compared with those of radial immunodiffusion assays. For comparison of the two systems, 232 patient sera were evaluated for C3, and 202 specimens were tested for C4. Analysis of the data by linear regression indicated a proportional difference between the methods. C3 and C4 concentrations measured by the fluorometric method were lower than those measured by radial immunodiffusion, especially concentrations exceeding the normal ranges. In detecting lower concentrations (less than 120 mg/dl for C3 and less than 25 mg/dl for C4), the two methods showed better agreement. Each assay system was reproducible and could be used to evaluate changes that occur in concentrations of complement components during therapeutic treatment. However, the ease in processing a large volume of specimens and the short time needed to complete the assay are advantages that make the fluorometric method more suitable than radial immunodiffusion for use in a large clinical laboratory.
Quantitation of rabbit immunoglobulin G antibodies to Salmonella minnesota Re by enzyme-linked immunosorbent assay
Authors: Eskenazy M , Naumova F , Tekelieva R , Konstantinov G
Published: January 2020

An enzyme-linked immunosorbent assay was developed which allowed the measurement of rabbit immunoglobulin G antibodies directed to Salmonella minnesota Re glycolipid. Efficient adsorption of the antigen to polystyrene could only be effected provided it had been previously dialyzed against 0.2 M EDTA (pH 7.0) and subsequently treated with 0.2% sodium deoxycholate (15 min at 56 degrees C) in 0.05 M diethanolamine buffer (pH 9.6). The method is by far more sensitive than quantitative precipitation in the determination of IgG antibodies. Inhibition by glycolipid of binding S. minnesota Re antibodies to immobilized glycolipid attests to the specificity of the assay and permits the detection of as little as 100 ng of glycolipid.
Evaluation of the macro-vue latex agglutination test for quantitation of gentamicin in human serum
Authors: Johnson J E , Crawford S , Jorgensen J H
Published: January 2020

The Macro-Vue Card Test (Hynson, Wescott, and Dunning, Baltimore, Md.) for rapid quantitation of gentamicin in serum was compared with bioassay and radioimmunoassay procedures on sera from 100 patients. Regression analysis of paired results from the bioassay (using Klebsiella pneumoniae ATCC 27799) and the Macro-Vue Card Test indicated a correlation coefficient of 0.89, whereas comparing radioimmunoassay and Card Test results yielded a correlation coefficient of 0.83. The bioassay-radioimmunoassay correlation coefficient was 0.88. The data were further examined by grouping the sera of the patients into three categories based on therapeutic concentration ranges as follows: less than 2 micrograms/ml, subtherapeutic range; 2 to 8 micrograms/ml, therapeutic range; greater than 8 micrograms/ml, potentially toxic range. Of 100 specimen values, 81 fell into the same concentration range when bioassay and Card Test values were compared. Of the 19 disagreements, 12 were considered minor. Of 100 specimen values compared by radioimmunoassay and the Card Test, 88 fell into the same concentration range. Of the 12 disagreements, 6 were considered minor. Therefore, the Macro-Vue Card Test for gentamicin compared favorably with both the bioassay and radioimmunoassay procedures. The minimal time required to perform the Card Test (12 to 15 min) makes it attractive for situations in which immediate results are needed.
Enzyme-linked immunosorbent assay for detection of respiratory syncytial virus infection: application to clinical samples
Authors: McIntosh K , Hendry R M , Fahnestock M L , Pierik L T
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) for respiratory syncytial virus antigens was applied to the rapid diagnosis of acute infections in children and was compared with viral culture and immunofluorescence tests. The ELISA test employed commercially available reagents and was run on a day-to-day basis as specimens were received in the laboratory. Sensitivity and specificity by ELISA were 82 and 95%, respectively, compared with culture. In the same specimens, the sensitivity and specificity by immunofluorescence were 86 and 96%, respectively. Nasopharyngeal aspirates were proven to be a better source of viral antigen than were nasopharyngeal swabs. ELISA-positive samples remained positive even when left unrefrigerated for a week or mailed to the laboratory in plastic containers. Respiratory syncytial virus ELISA, like culture, became negative as the disease progressed and showed no superiority over culture for diagnosis late in the illness.
Detection of immunoglobulin M antibody to Epstein-Barr virus by use of an enzyme-labeled antigen
Authors: Schmitz H
Published: January 2020

Immunoglobulin M (IgM) antibodies to Epstein-Barr virus were detected by using microtiter plates coated with anti-mu-chain antiserum and enzyme-labeled Epstein-Barr virus antigen. Optimum conditions for labeling were determined. The addition of unlabeled control antigen to the enzyme-labeled antigen was effective in reducing background reactions. Rheumatoid factor no longer interfered. Blocking of specific IgM antibody by IgG also was no longer observed. Four different methods for the detection of acute Epstein-Barr virus infections were compared. A combination of the enzyme-labeled antigen-IgM test with the detection of antibodies to either Epstein-Barr nuclear antigen or heterophile antigen was highly sensitive and specific. By changing the solid phase, IgA antibodies to Epstein-Barr virus could be detected.
Abnormal morphology of bacteria in the sputa of patients treated with antibiotics
Authors: Lorian V , Waluschka A , Kim Y
Published: January 2020

Filaments of Klebsiella pneumoniae were observed by Gram stain in the sputum of a patient with a respiratory infection who was treated with half the usual dose of cefazolin. Identical filaments were observed in vitro when this strain was incubated with subminimum inhibitory concentrations of cefazolin. Large gram-positive cocci containing multiple cross walls were observed by electron microscopy in the sputum of a patient with a respiratory infection who was treated with ampicillin and gentamicin. Antibiotic administration was suspended the night before the sputum was obtained. The ultrastructure of these cocci was very similar to the ultrastructure of Staphylococcus aureus incubated with subminimum inhibitory concentrations of cephaloridine or oxacillin. It was suspected that the low dose of cefazolin and the intermittent therapy with ampicillin resulted in a subminimum inhibitory concentration of antibiotic in the respiratory tract which induced the the abnormal morphology of the bacteria observed in the sputum of both patients. The presence of abnormal forms of bacteria in the specimen of a patient, rather than in the culture of a specimen, has clinical significance.
Isolation of
Campylobacter fetus
subsp.
jejuni
from Human Fecal Specimens by Incubation at 35 and 42°C
Authors: Janssen David , Helstad Arlan G.
Published: January 2020

Campylobacter
strains were isolated from human stool specimens that were cultured on modified Skirrow agar and incubated at 35 and 42°C. Incubation at 42°C resulted in more isolates, increased colony numbers, and earlier isolation than was found at 35°C.
Isolation of Mycobacterium chelonei with the lysis-centrifugation blood culture technique
Authors: Fojtasek M F , Kelly M T
Published: January 2020

Mycobacterium chelonei was isolated from a patient by the lysis-centrifugation and the conventional two-bottle blood culture methods. The lysis-centrifugation method was significantly more sensitive and rapid than the conventional method in detecting and isolating this organism; quantitations done by this method were useful for monitoring response to therapy.
Rotavirus gastroenteritis and weather
Authors: Brandt C D , Kim H W , Rodriguez W J , Arrobio J O , Jeffries B C , Parrott R H
Published: January 2020

During 5.5 years of a study in Washington, D.C., hospitalizations of children for rotavirus gastroenteritis tended to be more common after a month of cold or dry weather than after a corresponding calendar month of warm or wet weather. Overall, there were 84% more (178 versus 97) inpatients with rotavirus gastroenteritis after a set of relatively colder individual months taken as a group than after an equal number of warmer corresponding calendar months taken as a group. Comparable differences were not seen with nonrotavirus gastroenteritis patients. There also were 45% more rotavirus hospitalizations after the set of months with the least depth of precipitation compared with the set of corresponding calendar months with the greatest depth of precipitation. Rotavirus infection in young infants, the children least likely to be directly exposed to outdoor conditions, showed some of the most marked weather-associated effects. These findings suggest that weather-related low indoor relative humidity and indoor crowding may be key factors in the epidemiology of rotavirus disease.
Identification of Staphylococcus species with the API STAPH-IDENT system
Authors: Kloos W E , Wolfshohl J F
Published: January 2020

The API STAPH-IDENT system was compared with conventional methods for the identification of 14 Staphylococcus species. Conventional methods included the Kloos and Schleifer simplified scheme and DNA-DNA hybridization. The API STAPH-IDENT strip utilizes a battery of 10 miniaturized biochemical tests, including alkaline phosphatase, urease, beta-glucosidase, beta-glucuronidase, and beta-galactosidase activity, aerobic acid formation from D-(+)-mannose, D-mannitol, D-(+)-trehalose, and salicin, and utilization of arginine. Reactions of cultures were determined after 5 h of incubation at 35 degrees C. Results indicated a high degree of congruence (greater than 90%) between the expedient API system and conventional methods for most species. The addition of a test for novobiocin susceptibility to the API system increased the accuracy of identification of S. saprophyticus, S. cohnii, and S. hominis, significantly. Several strains of S. hominis, S. haemolyticus, and S. warneri which were difficult to separate with the Kloos and Schleifer simplified scheme were accurately resolved by the API system.
Long-term Staphylococcus aureus carrier state in hospital patients
Authors: Zierdt C H
Published: January 2020

Staphylococcus aureus colonization of 326 predominantly chronic-care (long-term) patients was studied for 24 years. There were 5,827 upper respiratory cultures positive for S. aureus, ranging from 10 to 88 per patient, determined by the number of years (1 to 21; average, 4.6) that the patient was studied. Patients on the average carried 2.8 S. aureus strains. One patient carried eight strains. Each patient had a predominant strain. Predominant strains tended to be permanent, with 87% persistence over the studied years. Lytic group III strains were more than twice as frequent as group I strains and eight times as frequent as group II strains. Patient carriage of multiple S. aureus strains was usual. A collection of large numbers of cultures over an extended period was necessary for this statistical study, since cultures positive for predominant strains were interspersed with negative cultures and cultures positive for minor strains. Thus, persistence of carriage of a predominant S. aureus strain in a patient continues despite frequent negative cultures and cultures positive for minor strains.
Clinical laboratory comparison of a slide blood culture system with a conventional broth system
Authors: Pfaller M A , Sibley T K , Westfall L M , Hoppe-Bauer J E , Keating M A , Murray P R
Published: January 2020

The recovery of bacteria and fungi from blood cultures was compared in conventional tryptic soy broth (TSB) bottles and in TSB bottles with an agarcoated slide attachment. A total of 2,662 sets of blood cultures, including 413 that were positive (15.5%), were evaluated. Significantly more gram-positive and gram-negative bacteria were recovered in the slide culture bottles than in conventional bottles (299 versus 253 isolates). Growth of gram-positive organisms and fungi was detected in the slide culture bottles 24 to 48 h earlier than in the TSB bottles. In addition, 76% of the isolates in the slide culture system were detected on the agar slide. In comparison, only 40% of the isolates in the TSB bottles were detected initially by blind subculturing. The incidences of contamination were 2.7% (71 cultures) for the slide culture system and 1.5% (39 cultures) for the TSB bottles.
Sensitivity of passive bacterial agglutination for detection of hepatitis B surface antigen
Authors: Rajagopalan M S , John T J
Published: January 2020

The sensitivity of passive bacterial agglutination (PBA), i.e., the agglutination of Staphylococcus aureus coated with antibodies to hepatitis B surface antigen by hepatitis B surface antigen, was assessed by testing reference panel no. 3A sera (Bureau of Biologics, U.S. Food and Drug Administration). Of the 23 samples containing hepatitis B surface antigen, 18 were positive by PBA. Sera from 100 healthy adults were tested by PBA and reversed passive hemagglutination. Four sera were positive by both methods. One additional sample was positive only by PBA; if one assumes that it was a false-positive result, the rate of false-positive reactions was only 1%.
Incidence of rotavirus infection in different age groups of pediatric patients with gastroenteritis
Authors: Truant A L , Chonmaitree T
Published: January 2020

An enzyme immunoassay was used to detect rotavirus in the stools of 176 pediatric patients presenting with gastroenteritis. The highest incidence of rotavirus infection was found among patients less than 1 year of age. In contrast to previously reported studies, 23% of neonates with gastroenteritis had rotavirus in their stools. This relatively easy and rapid method was helpful in the management of pediatric patients with gastroenteritis.
Evaluation of a prereduced anaerobically sterilized medium (PRAS II) system for identification anaerobic microorganisms
Authors: Beaucage C M , Onderdonk A B
Published: January 2020

A prereduced, anaerobically sterilized system of tubed media (PRAS II; Scott Laboratories, Fiskeville, R.I.) was evaluated for accuracy in the identification of anerobic microorganisms. PRAS II was found to be a rapid and accurate identification system for obligate anaerobes which does not require the use of gas cannula inoculation or incubation in a special anaerobic environment.
Rapid Trichrome Stain
Authors: Flournoy D. J. , McNabb Scott J. N. , Dodd Everett D. , Shaffer Marilynne H.
Published: January 2020

A rapid trichrome stain procedure was evaluated. It saves considerable time, especially when only a few slides are stained, and allows for the accurate identification of amoebae and other fecal parasites.