bluuebunny/crossref_metadata_2025 huggingface dataset:
Epstein-Barr virus-specific serum immunoglobulin A as an acute-phase antibody in infectious mononucleosis
Authors: Nikoskelainen J , Neel E U , Stevens D A
Published: January 2020

Immunoglobulin A (IgA) antibodies to Epstein-Barr virus viral capsid antigen were assayed serially in 19 patients with infectious mononucleosis and in 38 controls. Seventy-four percent of infectious mononucleosis patients demonstrated IgA antibody, whereas this was found in 13% of controls. This antibody appeared early in infectious mononucleosis and was virtually gone 10 weeks after onset. Comparison of IgA antibody kinetics was made with IgG and IgM antibodies to viral capsid antigen, heterophile antibody, and antibody to Epstein-Barr virus early antigen and nuclear antigen. Failure to demonstrate IgA antibody was associated with severe illness, prolonged illness, delay in IgG and anti-Epstein-Barr virus nuclear antigen antibody, and low or absent heterophile and anti-early antigen antibody. Assay of IgA antibody to viral capsid antigen is a potentially useful adjunct in the serodiagnosis of infectious mononucleosis or recent Epstein-Barr virus infection, as are the other antibodies tested, but in this study IgM viral capsid antigen antibody was the only acute-phase antibody present in all patients.
Primary plate identification of group A beta-hemolytic streptococci utilizing a two-disk technique
Authors: Baron E J , Gates J W
Published: January 2020

A two-disk system is described which allows primary plate identification of group A beta-hemolytic streptococci. Group A beta-hemolytic streptococci could be visualized on primary throat culture plates by using trimethoprim-sulfamethoxazole to inhibit normal flora. In the heavily inoculated area of Trypticase soy agar plates containing 5% sheep blood, a 25-microgram/ml trimethoprim-sulfamethoxazole disk was placed contiguous to a 0.04-U bacitracin disk. A total of 259 throat specimens were examined with this two-disk system. The swabs from these throat specimens were incubated in Todd-Hewitt broth. The bacterial pellet from the broths was stained by fluorescent antibody as a control. Of the cultures that were determined to be positive on the plates, 75% could be read unequivocally after overnight incubation, whereas the remaining 25% required subculture. The plates recovered 91% of the cultures which were considered as true positives by the broth-fluorescent-antibody technique. This method provided a significant savings in time compared with standard plate methods and in cost of materials compared with broth-fluorescent-antibody methods. This technique is particularly valuable for producing rapid results in laboratories where fluorescence microscopy would not be cost-effective.
Characteristics of Noncultivable Adenoviruses Associated with Diarrhea in Infants: A New Subgroup of Human Adenoviruses
Authors: Gary G. William , Hierholzer John C. , Black Robert E.
Published: January 2020

Virus particles morphologically resembling adenovirus were found in fecal specimens from infants and were examined for cultivability with standard cell culture techniques and for characteristics of human adenoviruses. Specimens from 13 of 15 infants could not be cultivated in cell cultures. The two adenoviruses that were cultivated, types 1 and 31, reacted in the expected manner in all tests. Counterimmunoelectrophoresis with group-specific anti-hexon serum confirmed that the observed particles in the 15 specimens were human adenoviruses. The buoyant density in sucrose of five of the noncultivable adenoviruses in original stool suspensions averaged 1.335 g/cm
3
and that of the two cultivable ones averaged 1.332 g/cm
3
; both groups had typical adenovirus morphology by electron microscopy. Treatment of the specimens and of a variety of tissue culture cells with proteolytic and other enzymes did not improve cultivability. Examination of partially purified virus by immunoelectron microscopy did not reveal evidence of immunoglobulin A, G, or M coating on the particles, an indication that coproantibody inhibition was not the cause of noncultivability. Fluorescent-antibody studies with an antihexon conjugate and counterimmunoelectrophoresis studies of serially passaged noncultivable viruses indicated that the viruses are infecting cells but are not undergoing effective replication. Antisera to three of the noncultivable viruses demonstrated homologous reactions in counterimmunoelectrophoresis with the respective immunizing antigens but showed only low levels of hemagglutination-inhibiting and neutralizing activity to a few of the known human adenoviruses. We concluded that the noncultivable viruses in these infant diarrhea cases were indeed human adenoviruses, were not defective particles, were not bound to coproantibody, were infectious but incapable of effective relication in conventional cell cultures, were serologically related to types 11, 17, 32, and 33, and should be considered a new, distinct subgroup.
Teichoic acid antibody determination by agar-gel diffusion: effect of using dilute antigen preparations
Authors: Wheat L J , Kohler R B , White A
Published: January 2020

Because the reported frequency of teichoic acid precipitins in controls and various patient groups has varied considerably among laboratories, we studied the effect of using various concentrations of staphylococcal extracts in agar-gel diffusion tests for teichoic acid antibodies. Of 25 normal sera, only 1 was positive against an undiluted extract, but 4 were positive against a 16-fold-diluted extract. Of nine sera from patients with staphylococcal bacteremia, two were positive at a higher titer against the diluted extract. A false-positive serum against the undiluted extract had a twofold titer increase against the diluted extract. Because human immune serum globulin is generally used as a positive teichoic acid antibody control, the variability of five different lots was studied. Three lots ahd teichoic acid antibody titers of 1:4, whereas one each had titers of 1:2 and 1:8. Based on this study, we feel that staphylococcal extracts should not be diluted. If immune serum globulins are used to determine the adequacy of ultrasonic extracts, newly acquired globulin lots should be standarized against an ultrasonic extract of proven sensitivity and specificity.
Nonfermentative bacilli: evaluation of three systems for identification
Authors: Otto L A , Blachman U
Published: January 2020

Three systems for the identification of nonfermentative bacilli were evaluated for their rapidity and accuracy of identification of 217 strains. Two of the systems, API 20E (API) and Oxi/Ferm tube (OxiF), are available as kits; the oxidative attack (OA) system is not commerically available. The overall accuracies of the OA, API, and OxiF systems were 91, 69, and 50%, respectively. Identification within 48 h was achieved for 98% of the strains by OA, for 50% by API, and for 18% by OxiF. Most of the organisms that were either misidentified or not identified by API and OxiF were those nonfermentative bacilli which are relatively more fastidious or rarely encountered or both. All three systems accurately identified nonfermentative bacilli commonly isolated at Olive View Medical Center, namely, Pseudomonas aeruginosa, Acinetobacter anitratus, Pseudomonas maltophilia, Acinetobacter lwoffi, saccharolytic flavobacteria (CDC IIb), moraxellae, Pseudomonas fluorescens, and Pseudomonas putida. The OA system identified 100% of the above organisms correctly, API identified 99.4%, and OxiF identified 99.3%. Since these organisms comprise 92% of the total number of nonfermentative bacilli isolated at Olive View Medical Center, we conclude that both API and OxiF may be useful alternatives to conventional methods, based on accuracy of identification alone. These two systems were considered substantially inferior to the OA system when both accuracy and rapidity of identification were taken into account.
Airway colonization by Flavobacterium in an intensive care unit
Authors: du Moulin G C
Published: January 2020

A total of 195 patients admitted to a respiratory-surgical intensive care unit became colonized with species of Flavobacterium during a 70-month prospective study. By biochemical, cultural, and morphological criteria and a comparison of antibiotic susceptibilities, all patient isolates of Flavobacterium were apparently related. The origin of these organisms was sought. Flavobacterium were recovered from different water-associated areas of the hospital and from the hands of respiratory-surgical intensive care care unit staff. The organisms were also found in university dormitory sinks. The isolation of these organisms from tap water led to their recovery from reservoirs supplying drinking water to the city of Boston and surrounding communities. These organisms are resistant to chlorine concentrations found in municipal water. There was no proven case of pneumonia caused by Flavobacterium in 2,329 consecutive patients studied in our respiratory-surgical intensive care unit.
Biotypes of Haemophilus encountered in clinical laboratories
Authors: Oberhofer T R , Back A E
Published: January 2020

The biochemical characteristics of 464 strains of Haemophilus influenzae and 83 strains of Haemophilus parainfluenzae isolated over an 18-month period are described. Of 22 characteristics obtained, only 6 were necessary to biochemically identify and biotype the isolates. The key substrates or tests were urease, ornithine, indole, o-nitrophenyl-beta-D-galactopyranoside, sucrose, and xylose. Five biotypes of H. influenzae and four of H. parainfluenzae were commonly recognized. Some strains were encountered which could not be accommodated in the recognized taxa but which constituted separate biotypes of the two species, H. influenzae biotype I was recovered principally from blood, cerebrospinal fluid, and upper respiratory secretion, and biotypes II and III were recovered from eye and sputum cultures. Biotype I was recovered primarily from children less than 1 year of age, whereas biotypes II and III were from persons 1 to 5 years old and from those over 20 years of age. Multiple isolates recovered from the same patient were almost always of the same biotype. Strains of H. parainfluenzae were isolated primarily from sputum, with others being isolated from body sources such as dental abscesses, gastric aspirates, and peritoneal fluid. An inverse relationship was noticed between hemolysis and mannose fermentation among H. parainfluenzae biotype III strains, whereas the relationship was absent among the other biotypes.
Comparison of the API 20E and Corning N/F systems for identification of nonfermentative gram-negative rods
Authors: Warwood N M , Blazevic D J , Hofherr L
Published: January 2020

A total of 231 strains of nonfermenting gram-negative rods were tested on the API 20E system, the Corning N/F system, and conventional media. When the results of identification to species were compared, the API system agreed with the conventional system on 69% of the isolates, and the Corning system agreed with the conventional system on 79% of the isolates. Both kit systems were deficient in identifying Pseudomonas cepacia and the more unusual isolates.
Cellular fatty acid composition of Pseudomonas paucimobilis and groups IIk-2, Ve-1, and Ve-2
Authors: Dees S B , Moss C W , Weaver R E , Hollis D
Published: January 2020

The cellular fatty acid composition of Pseudomonas paucimobilis (IIk-1) and group IIk-2, Ve-1, and Ve-2 was determined by gas-liquid chromatography. The unnamed groups were readily distinguished from P. paucimobilis by cellular fatty acids. The data strongly suggest that these bacteria may be additional species of Pseudomonas and Flavobacterium.
Recognition of group B streptococci in dip-slide cultures of urine
Authors: Jokipii A M , Jokipii L
Published: January 2020

One hundred strains of group B streptococci isolated from human infections were tested for growth on dip-slides available for the culture of urine. All grew on CLED agar, and none grew on MacConkey agar. The colonies were barely or not at all visible to the naked eye after overnight incubation (diameter, around 0.1 mm). The colony size increased eith prolonged incubation, but not if the inoculum density exceeded 10(6)/ml. Differences were found between lots of dip-slides. Poor growth on dip-slides may explain why group B streptococci have received little attention as pathogens of the urinary tract. The dip-slide screening personnel of one laboratory were informed of the experimental findings, and they started the practice of frequent subculture and prolonged incubation. The proportion of group B streptococci in significant bacteriuria increased from 0 to about 2% of positive cultures, whereas there was no conmitant increase of group B streptococci in dip-slides screened in several other laboratories serving as controls.
Effects of cell culture and laboratory conditions on type 2 dengue virus infectivity
Authors: Manning J S , Collins J K
Published: January 2020

The stability of type 2 dengue virus to exposure to a variety of laboratory conditions was determined. Suckling mouse brain passage virus was adapted for growth in BHK-21 cells, and plaque assays were performed using a tragacanth gum overlay. A three- to fourfold increase in plaque size could be obtained if monolayers were subconfluent at time of inoculation. Incubation of virus for 24 h at 37 degrees C, pH 6.5, or in buffer containing 1 mM ethylenediaminetetraacetate considerably reduced virus infectivity as compared with virus incubated for the same period at 4 degrees C, pH 8.0, or in buffer with or without 1 mM CaCl2 and 1 mM MgCl2. Multiple freezing and thawing of virus tissue culture medium containing 10% fetal calf serum did not reduce virus infectivity.
Application of the Fortner principle to isolation of Campylobacter from stools
Authors: Karmali M A , Fleming P C
Published: January 2020

A simple biological technique for reducing oxygen tension which does not necessitate the use of conventional anaerobic equipment is described. We successfully applied this method to the isolation of campylobacters from stools.
Modified inoculum for the enteric Minitek system from positive blood cultures
Authors: Cleary T J , Valdes S , Selem M
Published: January 2020

A modified Minitek inoculum procedure for the identification of Enterobacteriaceae from positive blood cultures was shown to be reliable. The method consisted of inoculating the Minitek enteric and nonfermenter broth with blood culture fluid and incubating the inoculum for 4 h before use.
Counterimmunoelectrophoresis assay for detection of adenovirus antigen
Authors: Mankikar S D , Middleton P J , Petric M
Published: January 2020

A rapid, sensitive counterimmunoelectrophoresis assay was developed to detect adenovirus in stools of patients with gastroenteritis. The parameters of the assay were optimized, and its sensitivity and efficiency was examined. The assay promises to be a useful alternative to conventional techniques of cell culture isolation or electron microscopy for the detection of adenovirus in stool specimens.
Group B beta-hemolytic streptococci causing pharyngitis
Authors: Chretien J H , McGinniss C G , Thompson J , Delaha E , Garagusi V F
Published: January 2020

Group B beta-hemolytic streptococci were isolated from the throats of 49 of 1,110 patients who had pharyngitis. Compared with patients whose throat cultures were negative for beta-hemolytic streptococci, those harboring group B were more likely to have enlarged tonsils (P less than 0.001), exudate (P less than 0.02), and tender enlarged anterior cervical lymph nodes (P less than 0.01). Group B should not necessarily be dismissed as a nonpathogen when identified in the pharynx of patients with exudative pharyngitis; laboratories which report beta-hemolytic streptococcal isolates from the pharynx only as group A or non-group A should be encouraged to perform definitive group identification of all beta-hemolytic isolates to further evaluate the role of other streptococcal groups as the causative agents for pharyngitis.
Use of an immunoperoxidase method for identification of Bacteroides fragilis
Authors: Hsu P C , Minshew B H , Williams B L , Lennard E S
Published: January 2020

An indirect immunoperoxidase (IP) slide test was evaluated for the laboratory identification of Bacteroides fragilis. Antigen-antibody complexes were detected with goat anti-rabbit immunoglobulin G-peroxidase conjugate with 3-amino-9-ethyl-carbazole as the peroxidase substrate. Ninety-one percent of 44 B. fragilis strains tested were IP positive (3+ to 4+ reactions) with greater than or equal to 1:160 dilutions of rabbit antiserum produced against whole cells of B. fragilis ATCC 23745. The antiserum was species specific. No cross-reactions were observed with 35 Bacteroides strains of other species or with a variety of facultative or aerobic gram-negative bacilli. Four B. fragilis strains were IP negative. One of these (VPI 2393) was the deoxyribonucleic acid (DNA) homology group II reference strain. The other three were clinical isolates. IP-negative and representative IP-positive strains were tested for DNA homology with the type strains for DNA homology groups I and II (VPI 2553 and VPI 2393). Two of the three clinical isolates were classified as DNA homology group II, and the remaining strain was classified as a group I. Capsular material known to be unique to B. fragilis was common to both DNA homology groups as indicated by reactions with purified anticapsular antiserum. The IP technique provides a suitable alternative to fluorescent microscopy for the rapid immunological identification of B. fragilis.
Comparison of a slide coagglutination technique with the Minitek system for confirmation of Neisseria gonorrhoeae
Authors: Hampton K D , Stallings R A , Wasilauskas B L
Published: January 2020

The Phadebact gonococcus test was performed on 314 bacterial isolates to compare the coagglutination technique with the Minitek system for the confirmation of Neisseria gonorrhoeae. Direct testing, using colonies of N. gonorrhoeae mixed with the Phadebact gonococcus test reagents, produced noninterpretable results in many cases. However, an alternative method, described by the manufacturer, of heating several colonies in distilled water for 20 min before testing alleviated most interpretation difficulties. Complete agreement was achieved with both the Phadebact and Minitek tests for all 236 isolates of N. gonorrhoeae. Of the 78 nongonococcal isolates tested with Phadebact, two isolates gave false-positive results, whereas one isolate was reported as noninterpretable. The Phadebact gonococcus test appears to be a simple and rapid alternative to confirmatory biochemical procedures for N. gonorrhoeae.
Natural infection of dogs on Cape Cod with Rickettsia rickettsii
Authors: Feng W C , Murray E S , Rosenberg G E , Spielman J M , Waner J L
Published: January 2020

Four isolates of rickettsiae from sick dogs on Cape Cod, Mass., were serologically identical to isolates of Rickettsia rickettsii from human patients with Rocky Mountain spotted fever. The antigenic analysis used the indirect fluorescent-antibody test and antisera prepared in mice to each of the isolates and to reference strains of R. rickettsii and Rickettsia montana. Serological responses of infected dogs were specific for R. rickettsii, although antibodies to R. montana were also detected in the sera of most of the canines.
Quantitative assay of soluble beta-hemolytic streptococcal antigens via an immunochemical turbidimetric method with a spectrophotometer
Authors: Levinson S S
Published: January 2020

Soluble, group A, B, C, and G beta-hemolytic streptococcal antigens were successfully identified in a prototype spectrophotometric system by an immunochemical turbidimetric assay. Any spectrophotometic system which can take a zero reading followed by a second reading 2 or more min later can be used for the assay. Maximum absorbance was obtained near a wavelength of 340 nm. A wide range of linearity between antigen concentration and absorbance was observed at some antibody dilutions, resulting in a simple assay which can be used to quantitate amounts of antigen in solution. Minimal cross-reactions that present no problem in interpretation was observed. Simulated emergency samples were solubilized and assayed for group A and B bacteria within 3 h of colony recognition. Reproducibility of the absorbance resulting from the antibody-antigen reaction was great, with low coefficients of variation over a period of 50 days. The simplicity of the assay solutions, requiring only antisera and a buffer, and the accessibility to high levels of quality control are among the greatest assets of the technique to clinical laboratories.
Evaluation of the new API 20C strip for yeast identification against a conventional method
Authors: Land G A , Harrison B A , Hulme K L , Cooper B H , Byrd J C
Published: January 2020

The new API 20C yeast identification system together with appropriate microscopic morphology determinations achieved a 97% correlation with a rapid conventional method. Whereas a group composed of Candida, Torulopsis, Saccharomyces, and Rhodotorula was identified with ease (98% overall correlation), a second group, containing Cryptococcus, Trichosporon, and Geotrichum species, appeared to give the system the most difficulty (90% correlation). Within this group particular difficulty was encountered in identifying varieties of Cryptococcus albidus, C. terreus, C. laurentii, Trichosporon beigelli, and Geotrichum spp. as to species. The API 20C system should be incubated the full 72 h prescribed by the manufacturer. However, when used in conjunction with appropriate morphological tests, presumptive identifications of some Candida and Torulopsis species may be made at 24 to 48 h. To facilitate identifications of the more difficult group of yeasts, ancillary tests for determining nitrate reductase, urease, and phenol oxidase activities should be considered as additions to the strip. Incorporating the phenol oxidase test would be especially important for identification of Cryptococcus neoformans, a yeast which should be identified as quickly and as accurately as possible. The API 20C system with computer assistance has proved to be an easy-to-inoculate, versatile, and fairly rapid method of yeast identification, giving results comparable to those obtained by conventional methodologies.
New method for estimating digestion of Paracoccidioides brasiliensis by phagocytic cells in vitro
Authors: Goihman-Yahr M , Essenfeld-Yahr E , Albornoz M C , Yarzábal L , de Gómez M H , San Martín B , Ocanto A , Convit J
Published: January 2020

We describe a method by which phagocytosis and digestion of Paracoccidioides brasiliensis yeast cells by polymorphonuclear leukocytes or other phagocytic cells may be estimated. Suspensions of P. brasiliensis in its yeastlike phase were sonicated, counted, and incubated with known numbers of peripheral blood polymorphonuclear leukocytes. At given intervals, cytocentrifuge droplets were stained by a variation of Papanicolaou's method. Stained preparations were examined with phase-contrast optics. Digested organisms showed total or partial disappearance of protoplasm. Green-stained cell walls resisted digestion. The proportion of digested cells as a function of time was estimated.
Rapid selective urease test for presumptive identification of Cryptococcus neoformans
Authors: Zimmer B L , Roberts G D
Published: January 2020

A rapid simplified screening method that selectively detected the urease activity of 99.6% of 286 isolates of Cryptococcus neoformans within 15 min was developed for use in clinical microbiology laboratories.
Evaluation of anaerobic incubation for recovery of group A streptococci from throat cultures
Authors: Pien F D , Ow C L , Isaacson N S , Goto N T , Rudoy R C
Published: January 2020

No statistical differences were found in the recovery of group A streptococci from throat culture specimens after overnight incubation of blood agar plates in 5% CO2 compared with anaerobiosis. Anaerobic incubation required many more subcultures and resulted in considerably greater technical time and expense.
Compatible results obtained from biotyping and serotyping in Serratia marcescens
Authors: Grimont P A , Grimont F , Le Minor S , Davis B , Pigache F
Published: January 2020

The correspondence between complete serotype and biotype (P.A.D. Grimont and F. Grimont, J. Clin. Microbiol. 8:73-83, 1978) of 474 Serratia marcescens strains was studied. Of 127 serotypes, 70 were represented by two or more strains of the same serotype belonged to one biotype. However, for 91% of serotypes, strains of the same serotype belonged to one biogroup--i.e., a group of closely related biotypes. Biogroups are A1 (A1a, A1b); A2/6 (A2a, A2b, A6a, A6b); A3 (A3a, A3b, A3c, A3d); A4 (A4a, A4b); A5/8 (A5, A8a, A8b, A8c); and TCT (TCT, TT). Only two serotypes were composed of a mixture of pigmented and nonpigmented biogroups. Pigmented biogroups (A1 and A2/6) were otherwise differentiated from nonpigmented biogroups (A3, A4, A5/8, and TCT) by serotyping. Some biogroups preferentially occurred in some O serogroups: A4 in 01; A2/6 in O6, O8, and O14; and A3 in O9, O12, and O15. Three H serogroups were found to be biochemically homogeneous: H1, H7, and H20 were respectively and uniquely composed of biogroups A4, TCT, and A3. A square matrix of O versus H serogroups, with the corresponding biogroup for each O X H combination, was used for comparisons between O groups and between H groups. Identical patterns of biogroups were shown by serogroups O6, O8, and O14. Taxonomical, ecological, and practical consequences of these findings are discussed.
Yersinia intermedia: temperature-dependent bacteriocin production
Authors: Bottone E J , Sandhu K K , Pisano M A
Published: January 2020

Yersinia enterocolitica and related Yersinia species share many temperature-dependent biological attributes. The present report documents temperature-dependent production of a bacteriocin-like substance only among Yersinia strains which ferment L-rhamnose, raffinose, and melibiose and which have been tentatively designed Y. intermedia. When tested by the "lawn-spotting" technique at 25 and 37 degrees C, 7 of 15 Y. intermedia strains produced a bacteriocin only at 25 degrees C with a uniform spectrum of activity against 2 strains of Y. enterocolitica, 1 rhamnose-positive, raffinose-negative, melibiose-negative strain tentatively designated Y. frederiksenii, 8 Y. intermedia, and 3 sucrose- and acetylmethylcarbinol-negative yersinial isolates tested. Bacteriocin-like activity was not detected among the Y. enterocolitica, Y. frederiksenii, or sucrose-negative yersinial strains tested. The exclusive activity of the Y. intermedia antibacterial substance only against yersiniae and not against other representative Enterobacteriaceae tested supports the placement of these microorganisms within the genus Yersinia and further establishes the singularity of Y. intermedia.
Indirect Sandwich Enzyme-Linked Immunosorbent Assay for Rapid Detection of
Haemophilus influenzae
Type b Infection
Authors: Drow Doris L. , Maki Dennis G. , Manning Dean D.
Published: January 2020

We report the development and testing of an enzyme-linked immunosorbent assay with excellent sensitivity for the detection of
Haemophilus influenzae
type b (HI
b
) antigen in clinical specimens from patients with HI
b
meningitis. The assay, an indirect sandwich technique, uses polystyrene balls as a solid phase and an alkaline phosphatase-labeled goat anti-rabbit globulin conjugate. Specimens are incubated with polystyrene balls armed with burro anti-HI
b
antiserum, and recognition antibody is visualized by addition of alkaline phosphatase-labeled anti-globulin, together with the enzyme substrate
p
-nitrophenyl phosphate. Concentrations of antigen are determined from standard curves prepared by using purified HI
b
capsular antigen polyribophosphate. The assay reproducibly detects polyribophosphate at concentrations between 1 and 5 ng/ml. Cross-reactions have not as yet been encountered in simulated and authentic clinical specimens containing other species including
Escherichia coli, Klebsiella pneumoniae,
group B
Streptococcus, Pseudomonas aeruginosa, Streptococcus pneumoniae, Staphylococcus aureus, Neisseria meningitidis,
and
Listeria monocytogenes.
In preliminary tests with 11 spinal fluid specimens, 2 serum specimens, and 5 urine specimens from patients with culture-proved HI
b
meningitis, antigen was detected in all specimens in concentrations ranging from 1 to 7,000 ng/ml. Antigen was not detected in any of 62 clinical specimens which were culture negative for HI
b
, including 11 spinal fluid specimens from patients with bacterial meningitis caused by microorganisms other than HI
b
. The enzyme-linked immunosorbent assay technique described here is considerably simpler than radioimmunoassay and, based on concurrent tests with 14 positive clinical specimens, may be more sensitive than counterimmunoelectrophoresis. It seems, therefore, to hold considerable promise for clinical use in rapid detection of systemic HI
b
infections.
Identification of Clinical Isolates of Mycobacteria with Gas-Liquid Chromatography Alone
Authors: Tisdall Philip A. , Roberts Glenn D. , Anhalt John P.
Published: January 2020

Identification of 18 mycobacterial species was performed by analysis of profiles obtained by using gas-liquid chromatography. Organisms were saponified in methanolic NaOH, and the reaction mixture was treated with BF
3
in methanol and extracted with a hexane-chloroform mixture. An identification scheme was developed from 128 stock strains and tested against a collection of 79 clinical isolates. By using gas-liquid chromatographic profiles alone, 58% of specimens were correctly identified to species level, and an additional 41% were correctly identified to a group of two or three organisms. Use in a clinical laboratory over a 2-month period proved chromatography to be as accurate as and more rapid than concurrent biochemical testing. Of 81 isolates tested, 64% were identified to species level by chromatography alone. An additional 35% were differentiated to the same groups of two or three organisms as found in our analysis of stock strains. These groups consisted of:
Mycobacterium tuberculosis, M. bovis
, and
M. xenopi; M. avium
complex,
M. gastri
, and
M. scrofulaceum
; or
M. fortuitum
and
M. chelonei.
Identification to species level from these groups could usually be done by colonial morphology alone and could always be done by the addition of one selected biochemical test. This study demonstrated the practical application of gas-liquid chromatography in the identification of mycobacteria in a clinical laboratory. In particular, all strains of
M. gordonae
and
M. kansasii
were identified to species level.
M. tuberculosis
was definitively identified in 85% of cases. When it could not be definitely identified, the only alternatives were
M. bovis
and
M. xenopi
, both of which are rare causes of infection.
Stability of Anti-
Candida
Precipitating Antibody in Stored Sera as Determined by Counterimmunoelectrophoresis
Authors: Dee Thomas H. , Berger Carol S.
Published: January 2020

Prolonged storage of sera at −20°C did not significantly affect anti-
Candida
precipitin titers measured by counterimmunoelectrophoresis when compared with the original determinations.
Colonization and infection of newborn infants caused by bacteriophage-group II Staphylococcus aureus strains
Authors: Blouse L E , Brockett R M , Steele N P , Ward E R
Published: January 2020

A staphylococcal strain which exhibited weak lytic reaction with group II phages was isolated from a newborn infant with a skin infection. Subsequent investigations established that this weakly reacting strain was responsible for an endemic level of infection and colonization within the hospital nursery. The use of consistently appearing weak lytic reactions in the evaluation of this episode is described.
Enzyme-Linked Immunosorbent Assay for Immunological Diagnosis of Human Tularemia
Authors: Carlsson Hans Erik , Lindberg Alf A. , Lindberg Gunilla , Hederstedt Bengt , Karlsson Karl-Axel , Agell Bengt O.
Published: January 2020

The enzyme-linked immunosorbent assay (ELISA) was applied for immunological diagnosis of human tularemia, using lipopolysaccharide from
Francisella tularensis
as antigen. Sera collected from patients, healthy individuals, and vaccinated volunteers were investigated for antibodies against
F. tularensis
by ELISA and tube agglutination. In ELISA all sera were titrated with a polyspecific anti-immunoglobulin enzyme conjugate. A limited number of consecutive sera from individual patients were also investigated for immunoglobulin G (IgG) and IgM antibodies by means of immunoglobulin class-specific conjugates. On an average ELISA was more than 10-fold as sensitive as tube agglutination. Two weeks after onset of disease, sera from patients had significantly higher titers in ELISA than sera from healthy controls. High titers persisted after more than 2 years. Significant amounts of both IgG and IgM antibodies were present within 1 to 2 weeks after infection. The antibody activity increased during the first month, without any significant change of the relation between IgG and IgM titers. After 2.5 years the IgG/IgM titer ratio of sera from patients was significantly increased. Within 6 weeks after vaccination sera from about half of the vaccinees had significantly elevated titers in ELISA. Titers observed after vaccination were generally lower than those found after infection. An elevated ELISA titer can be of diagnostic importance by the end of the first week of illness. A significant increase of titer in consecutive serum samples indicates a diagnosis of tularemia. Determination of IgG and IgM antibodies may be of value in determing whether a positive titer of a single serum sample is of longstanding or recent origin.
Biochemical characterization of unidentified microaerophilic cocci isolated from heifer and dry-cow mastitis
Authors: Schwan O , Nord C E , Holmberg O
Published: January 2020

Thirty-nine strains of gram-positive microaerophilic cocci isolated from cases of heifer and dry-cow mastitis were biochemically characterized with the API 50E and API-ZYM test kit systems, gas-liquid chromatography for analysis of end products of glucose metabolism, and anaerobic biochemical tests (L. V. Holdeman, E. P. Cato, and W. E. C. Moore, Anaerobe Laboratory Manual, Virginia Polytechnic Institute, Blacksburg, 1977). Strains were screened for production of a variety of extracellular enzymes on substrate-containing agar plates and for hemolysin and coagulase production. Antibiotic susceptibility and sensitivity tests were also performed. The microaerophilic cocci displayed homogeneity with respect to the majority of the biochemical tests used; i.e., greater than or equal to 90% of the strains were consistently positive or negative in any one test and probably represent one species. All produced deoxyribonuclease, ribonuclease, and hyaluronidase, and 92% were positive for chondroitin sulfatase. Catalase and coagulase tests were negative. Greening was observed on bovine blood agar. Acetic and succinic acids were produced by all strains as the only detectable products of glucose metabolism. The strains were susceptible to penicillin G, cefoxitin, doxycycline, and chloramphenicol and were resistant to clindamycin, novobiocin, and metronidazole. Their taxonomic position remains unclear.
Detection of immunoglobulin M antibodies to hepatitis A virus by enzyme-linked immunosorbent assay
Authors: Møller A M , Mathiesen L R
Published: January 2020

An enzyme-linked immunosorbent assay for the detection of immunoglobulin M (IgM) antibodies to hepatitis A virus is described. The test uses the principle of binding of IgM antibodies to anti-IgM-coated microtiter plates to determine whether the IgM antibodies attached have specificities for hepatitis A virus. In three patients with hepatitis type A followed up to 12 months, IgM antibodies to hepatitis A virus could be demonstrated from the onset of illness and during the following 2 to 3 months. When acute-phase sera from 48 patients with acute hepatitis were tested, IgM antibodies to hepatitis A virus could only be demonstrated in 18 patients previously classified as type A, whereas 30 patients with type B and non-A non-B hepatitis were negative. IgM antibodies to hepatitis A virus could not be demonstrated in 108 normal sera nor in 55 sera containing rheumatoid factor. These results indicate that the enzyme-linked immunosorbent assay for IgM antibodies to hepatitis A virus is useful in the serodiagnosis of acute hepatitis type A on a single serum sample taken during the acute phase of illness.
Evaluation of a biphasic medium for blood cultures
Authors: Hall M M , Mueske C A , Ilstrup D M , Washington J A
Published: January 2020

A study comparing the recovery of microorganisms from a transiently vented biphasic brain heart infusion medium bottle and a vacuum bottle containing tryptic soy broth demonstrated that growth was initially detected on the slant of the biphasic bottle or on a routine subculture of either broth in nearly 50% of instances. All organisms were isolated equally well in both bottles, with the exception of Staphylococcus aureus, which was isolated significantly more frequently from the biphasic bottle, and anaerobic bacteria, which were isolated significantly more frequently from the tryptic soy broth bottle.
Measure of immunoglobulin G-, M-, and A-specific titers against Legionella pneumophila and inhibition of titers against nonspecific, gram-negative bacterial antigens in the indirect immunofluorescence test for legionellosis
Authors: Wilkinson H W , Farshy C E , Fikes B J , Cruce D D , Yealy L P
Published: January 2020

A crude extract of Escherichia coli O13:K92:H4 inhibited 97% of positive indirect immunofluorescence titers against a variety of gram-negative bacterial antigens while lowering Legionella pneumophila titers in only 6% of sera from patients with suspected legionellosis. Legionella-specific titers were the result of immunoglobulins G, M, and A, singly or in combination.
Occurrence of protein A in Staphylococcus aureus and closely related Staphylococcus species
Authors: Lachica R V , Genigeorgis C A , Hoeprich P D
Published: January 2020

All but 1 of 143 strains of Staphylococcus aureus were positive for protein A, whereas all 34 strains of Staphylococcus hyicus and 123 of 127 strains of Staphylococcus intermedius were devoid of this cell wall component.
Relative recovery of anaerobes on different isolation media
Authors: Sondag J E , Ali M , Murray P R
Published: January 2020

The recovery of clinical anaerobic isolates on selective and nonselective agar media, as well as the time required to detect the isolates, was examined. Of a total of 235 isolates, 77, 46, and 40% were detected on Schaedler blood agar, colistinnalidixic acid blood agar, and kanamycin-vancomycin-lysed blood agar, respectively, and 94% were detected on the combination of Schaedler blood agar with kanamycin-vancomycin-lysed blood agar. A total of 19% of the anaerobes were detected after incubation for 1 day, and 70% were detected after 2 days.
O-serotyping Providencia alcalifaciens
Authors: Penner J L , Fleming P C , Whiteley G R , Hennessy J N
Published: January 2020

The O-serotyping scheme for Providencia was tested on Providencia alcalifaciens isolates collected mostly from two hospitals. The specificites of the somatic (O) antigens of P. alcalifaciens were found to be different from those of Providencia stuartii, and separation of the Providencia typing scheme to allow separate typing of each species led to more efficient typing. All but 4 of 86 isolates were typable. Eighteen serotypes occurred among 53 typable isolates obtained from a pediatric hospital, and 11 occurred among 19 isolates from a general hospital. Thirty-two percent of the isolates from the pediatric hospital belonged to serotype O3, the most frequently isolated and most widely distributed type. The use of the serotyping scheme for P. alcalifaciens is advocated for further studies to examine strains of the species for enteropathogenic types.
Evaluation of a ganglioside immunosorbent assay for detection of Escherichia coli heat-labile enterotoxin
Authors: Bäck E , Svennerholm A M , Holmgren J , Möllby R
Published: January 2020

The GM1 ganglioside enzyme-linked immunosorbent assay (GM1-ELISA), an immunological method for detection of Escherichia coli heat-labile enterotoxin (LT), was quantitatively and qualitatively compared with the conventional adrenal cell test for the identification of LT-producing strains. A micromodification model of the assay was developed. Enterotoxin preparations from 120 E. coli isolates from individuals with diarrhea, which had been previously shown to be enterotoxigenic by the adrenal cell test, and from 44 control strains of E. coli were compared in parallel by the two methods. Quantitatively the covariation of the enterotoxin titers was highly significant (RS = 0.98, P less than 0.001), the GM1-ELISA being somewhat more sensitive than the adrenal cell test. The methodological error was less than 5% in both tests. Qualitatively the overall agreement for positive and negative reactions for the two methods was 89%. The GM1-ELISA is practical for routine use in the diagnosis of enterotoxigenic E. coli, especially in laboratories without facilities for cell culture.
Identification of diaminopimelic acid in the Legionnaires disease bacterium
Authors: Guerrant G O , Lambert M S , Moss C W
Published: January 2020

Diaminopimelic acid was found to be a component of the cell wall of the Legionnaires disease bacterium, thus providing additional evidence that the organism is a bacterium. The presence of this amino acid was determined by gas-liquid chromatography and confirmed by gas chromatography-mass spectrometry.
Effect of storage temperature on viability of Trichophyton mentagrophytes in infected guinea pig skin scales
Authors: Sinski J T , Wallis B M , Kelley L M
Published: January 2020

The effect of storage temperature on the viability of Trichophyton mentagrophytes in infected guinea pig skin scales was quantitatively evaluated for 182 days. Infected skin scales were stored, while protected from light, in glass containers with desiccant at four temperatures and sampled after saline treatment at various time intervals. No decrease in viability was observed for storage at 4 degrees C, room temperature, or 30 degrees C. Storage at 37 degrees C resulted in a significant loss in viability.
Medium for selective isolation of Fusobacterium nucleatum from human periodontal pockets
Authors: Walker C B , Ratliff D , Muller D , Mandell R , Socransky S S
Published: January 2020

A selective medium, CVE agar, was developed for the isolation of Fusobacterium nucleatum from subgingival plaque of periodontally diseased patients. The medium contained 1.0% Trypticase (BBL Microbiology Systems), 0.5% yeast extract, 0.5% NaCl, 0.2% glucose, 0.02% L-tryptophan, 1.5% agar, and 5% defibrinated whole sheep blood. Erythromycin and crystal violet were added as the selective inhibitory agents at concentrations of 4 and 5 micrograms/ml, respectively. The medium permitted almost total recovery of F. nucleatum when compared with a nonselective medium and suppressed the recovery of most remaining species by nonselective medium and suppressed the recovery of most remaining species by 6 to 8 orders of magnitude. Microorganisms suppressed to a lesser degree included Selenomonas sputigena, Actinobacillus actinomycetemcomitans, Eikenella corrodens, and some strains of Peptostreptococcus. The distinct colonial morphology of F. nucleatum on CVE agar made differentiation relatively easy when contaminants were present. With this medium, F. nucleatum was enumerated from 278 subgingival plaque samples and accounted for less than 1.0 to greater than 25% of the cultivatable microbiota.
Microbiology subsystem of a total, dedicated laboratory computer system
Authors: Lawrie D J , Elin R J , Gill V J , Lewis T L , MacLowry J D , Witebsky F G
Published: January 2020

The computer system used by the Microbiology Service of the Clinical Pathology Department, Clinical Center, National Institutes of Health is discussed. This microbiology subsystem is a part of a dedicated on-line laboratory computer system used by the entire department. The laboratory computer is connected on-line to a hospital computer which provides patient admission, transfer, and discharge data. Mark sense worksheets and cathode ray tube terminals are used for result entry and correction. Cumulative patient reports are printed. Results for both active and completed accessions can be easily retrieved on cathode ray terminals in the laboratory. All laboratory data are archived on magnetic tape from which a research data base and microfiched laboratory records are generated. The manner in which the system is integrated in the routine operation of the microbiology laboratory is emphasized. In addition, some of the costs, benefits, liabilities, and pitfalls associated with the introduction of the computer in the laboratory are reviewed. Finally, we have presented our concept of some of the future enhancements to our present system and some of the directions in which any future microbiology system might develop.
Serology of Legionnaires disease: comparison of indirect fluorescent antibody, immune adherence hemagglutination, and indirect hemagglutination tests
Authors: Lennette D A , Lennette E T , Wentworth B B , French M L , Lattimer G L
Published: January 2020

An immune adherence hemagglutination (IAHA) test for the measurement of antibodies to Legionella pneumophila was developed and evaluated for the diagnosis of Legionnaires disease. Its sensitivity was compared to that of the indirect fluorescent antibody (IFA) test and a recently developed indirect hemagglutination (IHA) test. The sensitivity of the three tests appeared to be similar, with the IFA test giving slightly higher titers. Both the IHA and IAHA tests appear useful for the serodiagnosis of Legionnaires disease; the IAHA test has the advantage that it can be used with many other serological antigens.
Cultures for Clostridium difficile in stools containing a cytotoxin neutralized by Clostridium sordellii antitoxin
Authors: Willey S H , Bartlett J G
Published: January 2020

Stools from patients with antibiotic-associated diarrhea or colitis were cultured to detect the presence of Clostridium difficile. All specimens contained a cytotoxin which was neutralized by Clostridium sordellii antitoxin. Initial testing employed several methods with comparative merits in recovering this organism. These included the use of nonselective media, antibiotic-incorporated media, alcohol shock, and paracresol-containing broth. Optimal results were achieved with primary plating of serial dilutions onto a selective agar containing cycloserine and cefoxitin. This technique was then employed in a large number of specimens. The overall results showed that C. difficile was recovered in specimens from 71 of 73 patients. All isolates of C. difficile produced a cytotoxin which was neutralized by C. sordellii antitoxin in vitro. These results verify the utility of this medium and support the concept that C. difficile accounts for the cytotoxin found in stools in nearly all cases.