bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of antibodies to Epstein-Barr virus capsid antigen by immune adherence hemagglutination
Authors: Lennette E T , Ward E , Henle G , Henle W
Published: January 2020

The immune adherence hemagglutination assay was found to be as sensitive and specific as the indirect immunofluorescence technique for titration of antibodies to Epstein-Barr virus capsid antigen. Satisfactory virus capsid antigen-specific and negative control antigens for the immune adherence hemagglutination assay were prepared from cell extracts of the Epstein-Barr virus producer P3HR-1 and the Epstein-Barr virus genome-negative BJAB lymphoblastoid cell lines, respectively. As the immune adherence hemagglutination assay can be used to titrate antibodies to both the heterophil antigen of the Paul-Bunnell type and to virus capsid antigen, it offers a promising alternative to the immunofluorescence methods in the serodiagnosis of Epstein-Barr virus infections which can be performed by most diagnostic laboratories.
Lysis-filtration blood culture versus conventional blood culture in a bacteremic rabbit model
Authors: Zierdt C H , Peterson D L , Swan J C , MacLowry J D
Published: January 2020

Thirteen representative pathogenic bacterial species were used to create septicemia in rabbits, by injecting 10(6) colony-forming units into the marginal ear vein. At a selected time, usually 30 to 60 min after injection, heart blood was drawn into heparin and dispensed in 5.0-,0.5-, and 0.1-ml volumes into duplicate bottles of commercial brain heart infusion broth with sodium polyanetholesulfonate, and into duplicate bottles of a newly developed blood-lysing solution. Lysed blood was filtered, and the filter membranes were cultured in brain heart infusion broth. At the 5.0-ml blood inoculum level, of 126 total culture bottles (63 rabbits) for each system, 83 conventional cultures versus 109 lysis-filtration cultures were positive. At the 0.5-ml blood inoculum, 20 of 126 conventional culture bottles were positive, versus 66 of 126 lysis-filtration cultures. At the 0.1-ml blood inoculum, 2 of 126 conventional culture bottles were positive, versus 30 of 126 lysis-filtration cultures. Overall, 105 of 378 conventional cultures and 205 of 378 lysis-filtration cultures were positive. The advantage of the lysis-filtration system was striking for both gram-positive and gram-negative organisms at all inoculum concentrations, but was greater for gram-positive organisms. Most significant was the rate of recovery by this new system, when the number of bacteria in the blood was reduced to the point where recovery by conventional culture was unlikely. It is postulated that the superiority of lysis-filtration culture may be due to release of bacteria by lysis of phagocytes, preventing continued loss of pathogens by intracellular destruction during the first hours of blood culture.
Prevalence of maltose-negative Neisseria meningitidis variants during an epidemic period in Spain
Authors: Sáez-Nieto J A , Fenoll A , Vazquez J , Casal J
Published: January 2020

We studied the prevalence of maltose-negative variants of Neisseria meningitidis in Spain from 1978 to 1980. Sugar utilization studies were performed with both CTA medium and Mueller-Hinton medium; bromothymol blue was used as the indicator in Mueller-Hinton medium. Of 1,714 isolates of N. meningitidis recovered from the cerebrospinal fluid or blood of patients with meningococcal infections, 64 (3.7%) were maltose-negative variants; 13 (3.3%) of the 363 isolates found in carriers had the same characteristic. All maltose-negative cultures isolated from both patients and carriers belonged to serogroup B and were resistant to sulfadiazine at a minimal inhibitory concentration, 10 micrograms/ml or more. Serotype 2 isolates were the most prevalent isolates in patients (68.8%), followed by nontypable isolates (20.3%). Only serotype 2 isolates (66%) and nontypable isolates (33%) were found in carriers.
API ZYM system for identification of Bacteroides spp., Capnocytophaga spp., and spirochetes of oral origin
Authors: Laughon B E , Syed S A , Loesche W J
Published: January 2020

A total of 80 oral strains of Bacteroides gingivalis, B. asaccharolyticus, B. melaninogenicus subsp. intermedius, B. melaninogenicus subsp. melaninogenicus, Capnocytophaga, Treponema denticola, and T. vincentii were characterized with the API ZYM system for 19 enzyme activities. Comparison of anaerobic and aerobic incubation with nine reference strains of these organisms showed no important differences. The key differential tests for black-pigmented Bacteroides strains and treponemes of oral origin were trypsin, alpha-glucosidase, and N-acetyl-beta-glucosaminidase. All Capnocytophaga strains produced distinctive aminopeptidase activities but varied in their glycosidic capabilities. The presence of a trypsin-like activity in B. gingivalis, T. denticola, and a group of Capnocytophaga strains may contribute to tissue destruction in periodontal disease.
Influence of humidity on rotavirus prevalence among Nigerian infants and young children with gastroenteritis
Authors: Paul M O , Erinle E A
Published: January 2020

Rotaviruses were detected by counterimmunoelectro-osmophoresis in the feces of 16 (13.8%) of 116 infants and young children with gastroenteritis during a 5-month period (September 1979 through January 1980) in Ife, Nigeria. The rate of rotavirus detection varied inversely with relative humidity and was highest in December (38.5% positive) when the humidity was lower. There was not such a distinct relationship with temperature or vapor pressure; and although the rate of rotavirus detection was higher in the drier months (November to January; 19.3% positive) than in the rainy season months (September and October; 8.5% positive), the average humidity was lower in the drier months than in the rainy months. Low relative humidity (49 to 78%) is apparently the most important environmental factor for rotavirus survival and spread in this area.
Reliability of the MS-2 system in detecting methicillin-resistant Staphylococcus aureus
Authors: Boyce J M , White R L , Bonner M C , Lockwood W R
Published: January 2020

The MS-2 system (Abbott Diagnostics, Division of Abbott Laboratories, Dallas, Tex.) is an automated system capable of rapid antimicrobial susceptibility testing. However, the short incubation periods used by the device may adversely affect its ability to detect slowly growing resistant organisms. Shortly after the introduction of the MS-2 system into the University of Mississippi Medical Center clinical microbiology laboratory, we noted discrepancies between the MS-2 and the disk diffusion susceptibility reports when methicillin-resistant Staphylococcus aureus isolates were tested. Subsequently, we determined the susceptibilities of 75 such isolates by the MS-2 and Kirby-Bauer disk diffusion methods and measured the minimum inhibitory concentrations of methicillin, oxacillin, and cephalothin for 33 of the 75 isolates by standardized agar dilution techniques. There was only 47% overall agreement between the MS-2 and disk diffusion methods when methicillin was tested and 15% agreement when cephalothin was the test drug. There was 93% or more overall agreement between the two methods when other antimicrobial agents were tested. The minimum inhibitory concentration of methicillin was greater than or equal to 16 micrograms/ml for all 33 isolates evaluated by the agar dilution method. A comparison of the MS-2 and agar dilution results revealed an overall agreement of 49% when the susceptibilities to methicillin were determined. The MS-2 system reported that multiple methicillin-resistant S. aureus isolates obtained from a single patient were either resistant, intermediate, or sensitive to methicillin. Inconsistent results were also obtained when a single isolate was tested simultaneously in 10 cuvette cartridges. We conclude that the MS-2 system does not reliably detect methicillin and cephalothin resistance among S. aureus.
Evaluation of the Phadebact Test for identification of Neisseria gonorrhoeae
Authors: Carlson B L , Haley M S , Kelly J R , McCormack W M
Published: January 2020

The Phadebact Gonococcus Test (Pharmacia Diagnostics, Piscataway, N.J.) is used for the identification of Neisseria gonorrhoeae. In this test, boiled organisms are examined by using a 3-min coagglutination technique. A total of 313 isolates of Neisseria confirmed by the fluorescent-antibody technique or identified to the species level by the cysteine-tryptose agar utilization technique, were tested by the coagglutination technique. Of 229 isolates of N. gonorrhoeae, 13 (5.7%) were negative by the coagglutination technique on the first testing, 4 (1.7%) of which remained negative on subsequent testing after subculture. Of the 84 isolates of Neisseria other than N. gonorrhoeae, none showed a positive coagglutination reading. The Phadebact Gonococcus Test provides rapid, accurate identification for most isolates of N. gonorrhoeae. As with any new technique, standard procedures (fluorescent-antibody or cysteine-tryptose agar analysis or both) should be performed along with the new technique until laboratory workers are comfortable with the performance and interpretation of the test. We must, however, emphasize that it is very important to follow carefully the procedures outlined on the package insert.
Enzyme-linked immunosorbent assay for detection of antibody to Treponema hyodysenteriae antigens
Authors: Joens L A , Nord N A , Kinyon J M , Egan I T
Published: January 2020

The enzyme-linked immunosorbent assay (ELISA) was evaluated and compared with the microtitration agglutination test for the detection of swine antibody to Treponema hyodysenteriae lipopolysaccharide antigens. Cells of T. hyodysenteriae serotypes 1 and 2 were extracted with hot phenol-water (68 degrees C). The lipopolysaccharide fraction from the aqueous phase was coated on plastic wells at concentrations of 1 micrograms (serotype 1) and 10 micrograms (serotype 2) of carbohydrate per ml. The ELISA was serotype specific when lipopolysaccharide antigens were reacted against sera from convalescent swine. Seroconversion of infected pigs was detectable with the ELISA within 1 to 2 weeks postinoculation and with the microtitration agglutination test 2 to 3 weeks postinoculation. Antibody titers could be detected in convalescent pigs as long as 19 weeks postinoculation by the ELISA and 12 to 13 weeks postinoculation by the microtitration agglutination test. Therefore, the ELISA may be useful for the detection of asymptomatic carriers.
Rate of penicillin killing of Staphylococcus aureus and Autobac 1 susceptibility test results
Authors: Harris J A , Furtado D
Published: January 2020

A clinical isolate of Staphylococcus aureus interpreted as resistant to penicillin by the Autobac 1 susceptibility testing method (i.e., light-scattering index of 0.77) was found to be susceptible to penicillin by both the disk diffusion and broth dilution techniques. The growth rate of the clinical isolate during a 4-h incubation interval was similar to that of a known sensitive reference strain (S. aureus ATCC 25923) used as a control organism for the Autobac test. The bactericidal effect of penicillin was evaluated by measuring the rate of killing over a 4-h interval. The percentages of organisms surviving exposure to 5.0 or 2.5 U of penicillin G per ml (number of organisms recovered at 3 h/number of organisms introduced as inoculum) were 68 and 76%, respectively, for the clinical isolate and 15 and 21%, respectively, for the reference strain. After 24 h of incubation, penicillin was bactericidal for both strains. The need to increase the time of incubation for those S. aureus isolates resistant to penicillin after 3 h of standard incubation time in the Autobac system is discussed.
Analysis of short-chain acids from bacteria by gas-liquid chromatography with a fused-silica capillary column
Authors: Moss C W , Nunez-Montiel O L
Published: January 2020

The use of a flexible, fused-silica capillary column for gas-liquid chromatographic analysis of short-chain acids from bacteria is illustrated with a standard acid mixture and with a derivatized extract of culture medium from Clostridium difficile.
Use of chemotaxis chambers for studying in vitro bacterial colonization of biomaterials
Authors: Leake E S , Gristina A G , Wright M J
Published: January 2020

Blind-well chemotaxis chambers were used to study the in vitro bacterial adhesion and colonization of biomaterials. Staphylococcus aureus and Pseudomonas aeruginosa were selected for the bacterial inocula. Abundant growth on the surfaces of methyl methacrylate, polyethylene, stainless steel, and Vitallium was detected by using scanning electron microscopy after 24 h of incubation. The culture technique employed proved to be of value for the study of surface bacterial colonization of inert materials.
Simple immunodiffusion test for detecting antibodies against Legionella pneumophila serotype 1
Authors: Soriano F , Aguilar L , Gómez Garcés J L
Published: January 2020

A simple immunodiffusion test for investigation of antibodies against Legionella pneumophila serotype 1 was developed. The comparison between the indirect immunofluorescence assay and the immunodiffusion technique gave a 98.6% correlation. All of the patients studied were serologically diagnosed by both procedures.
Two rapid pigmentation tests for identification of Cryptococcus neoformans
Authors: Kaufmann C S , Merz W G
Published: January 2020

Two tests were developed for the rapid identification of Cryptococcus neoformans based on pigment produced by the organism's phenoloxidase activity. Caffeic acid was incorporated into cornmeal agar, a medium used routinely for yeast identification. When tested on this medium, only C. neoformans isolates produced brown pigment. All other yeasts maintained their normal morphology and did not produce the reaction product. A non-medium-based test was developed for same-day identification of C. neoformans isolates. Paper strips saturated with a buffered L-beta-3,4-dihydroxyphenylalanine-ferric citrate solution were inoculated with isolates and incubated at 37 degrees C. Pigment production occurred only with C. neoformans isolates, many within 60 to 90 min. All other yeasts remained negative.
Rapid Identification of
Bacteroides gingivalis
Authors: Laughon B. E. , Syed S. A. , Loesche W. J.
Published: January 2020

Rapid colorimetric tests for trypsin and α-glucosidase are described for use in the identification of
Bacteroides gingivalis
from dental plaque.
Exoantigen Studies of Sporothrix schenckii, Ceratocystis minor, and Graphium penicilliodes cultures
Authors: Polonelli L , Morace G
Published: January 2020

Cultures of Sporothrix schenckii were serologically tested by the exoantigen immunodiffusion technique of Kaufman and Standard (L. Kaufman and P.G. Standard, J. Clin. Microbiol. 8:42-45, 1978). This rapid and sensitive technique permitted the identification of 10 isolates of S. schenckii within 3 days. The production of the antigen-antiserum reference system and exoantigens with two different methods are reported. The demonstration of common antigens in S. schenckii and Ceratocystis minor, the suspected perfect state of S. schenckii, indicates that the two are antigenically related; however, the question as to whether C. minor represents the perfect form of S. schenckii will depend upon the induction of a sexual state in S. schenckii.
Rapid typing of herpes simplex virus based on immunological specificity of viral thymidine kinase and typing according to sensitivity to iododeoxyuridine
Authors: Gronowitz J S , Källander C F , Jeansson S , Wallin J
Published: January 2020

We describe two methods for typing of herpes simplex virus (HSV). One procedure is based on the finding that the multiplication of HSV type 1 strains in primary rabbit kidney cells is inhibited by 2 x 10(-5) M iododeoxyuridine, whereas growth of HSV type 2 strains is considerably less affected. Forty-nine different HSV isolates were typed according to this method. For all isolates except two the results were found to be in agreement with results obtained by another typing procedure, the counterimmunoelectroosmophoretic method (S. Jeansson, Appl. Microbiol. 24:96-100, 1972). One HSV type 1 isolate behaved as a type 2 strain and was found to be a deoxythymidine kinase-negative mutant strain. The other deviant strain exhibited an intermediate iododeoxyuridine sensitivity, thus being impossible to type with this method. Another, faster typing procedure which is based on the immunological difference between HSV type 1 and 2 deoxythymidine kinase is also presented. This assay, in combination with the conventional methods for isolation, enables the detection of deoxythymidine kinase-negative therapy-resistant HSV strains. Finally, we report the detection and typing of HSV deoxythymidine kinase present in vesicle fluids.
Comparison of the Oxi/Ferm and N/F systems for identification of infrequently encountered nonfermentative and oxidase-positive fermentative bacilli
Authors: Koestenblatt E K , Larone D H , Pavletich K J
Published: January 2020

The N/F system (Flow Laboratories, Inc. Rockville, Md) and the Oxi/Ferm Tube, (Roche Diagnostics, Div. Hoffman-La Roche, Inc. Nutley, N.J.) were evaluated in parallel for identification of infrequently encountered nonfermentative gram-negative bacilli and oxidase-positive, fermentative gram-negative bacilli selected from fresh clinical isolates and stock cultures. When compared with conventional media, the Flow N/F system correctly identified 97.7% (86 or 88) of all strains tested. No organisms were misidentified, but this system failed to identify two strains. Six supplemental tests were needed for the complete identification of 16 strains (18%); however, correct results were obtained within 48 h for 85% of the isolates. The Oxi/Ferm method correctly identified 87.5% of the isolates; 7% were incorrectly identified, and the method failed to identify five strains. Seventeen supplemental tests were required to identify 64 strains (73%). Complete identification within 48 h was obtained for 60% of the organisms tested.
Identification of Vibrio hollisae sp. nov. from patients with diarrhea
Authors: Hickman F W , Farmer J J , Hollis D G , Fanning G R , Steigerwalt A G , Weaver R E , Brenner D J
Published: January 2020

The name Vibrio hollisae (synonym = Special Bacteriology group EF-13) is proposed for a new group of 16 strains that occurred in stool cultures of patients with diarrhea. V. hollisae is a small gram-negative rod, which is motile with a single polar flagellum. No lateral or peritrichous flagella were observed, even when it was grown on a solid medium. Sodium chloride is required for growth, so V. hollisae is a halophilic vibrio. Strains were positive (36 degrees C, 24 or 48 h) for oxidase (Kovacs), indole production, nitrate reduction to nitrite, and fermentation of D-glucose (acid, no gas), L-arabinose, D-galactose, and D-mannose. Strains were negative for the following tests often used in enteric bacteriology: lipase (corn oil); deoxyribonuclease; gelatinase; methyl red; Voges-Proskauer; utilization of citrate, acetate, and malonate; L-lysine decarboxylase (Møllers); L-ornithine decarboxylase (Møllers); L-arginine dihydrolase (Møllers); growth in KCN medium; and acid production from D-adonitol, D-arabitol, cellobiose, dulcitol, erythritol, glycerol (25% delayed positive at 7 days), i-(myo)-inositol, lactose, maltose, D-mannitol, melibiose, alpha-methyl-D-glucoside, mucate, raffinose, L-rhamnose, salicin, D-sorbitol, sucrose, trehalose, and D-xylose. None of the strains was motile (semisolid medium) at 36 degrees C at 48 h, but by 7 days 88% were motile. The strains did not grow within 2 days when plated on thiosulfate-citrate-bile salts-sucrose (TCBS) agar or MacConkey agar, but they grew on sheep blood agar and marine agar. By DNA-DNA hybridization (75 degrees C, hydroxyapatite with (32)P), V. hollisae was only 0 to 4% related to 21 named species in Vibrio and Photobacterium. The type strain is designated ATCC 33564, which has a mean guanineplus-cytosine content in DNA of 50 mol%. With the disk diffusion method V. hollisae had relatively large zones of inhibition around penicillin, ampicillin, carbenicillin, cephalothin, colistin, polymyxin B, streptomycin, kanamycin, gentamicin, tetracycline, chloramphenicol, and sulfadiazine. Future studies should focus on the isolation of this new vibrio and its ecology and relationship to human diseases.
Rapid serological technique for typing herpes simplex viruses
Authors: Cleveland P H , Richman D D , Oxman M N , Worthen D M
Published: January 2020

A rapid technique is described which can accurately identify a herpes simplex virus (HSV) isolate as type 1 or type 2. Filter paper disks were used to immobilize viral antigens, which were then identified by means of an (125)I-labeled staphylococcal protein A immunoassay. The assay was performed in a specially designed 96-well filtration device which served as both an incubation chamber and a filter manifold. By using this system and cross-absorbed antisera to HSV types 1 and 2, 69 coded clinical isolates of HSV were correctly and unequivocally typed. HSV was also clearly distinguished from varicella-zoster virus and cytomegalovirus. This assay can be rapidly executed (less than 2 h) and yielded an objective endpoint; it required only minute quantities of typing sera and can be easily performed with the cells from a single infected roller tube culture. Thus, it can be used to type initial clinical isolates of HSV, yielding results within hours after the first appearance of cytopathic effects in the culture used for primary virus isolation. Moreover, it is particularly well suited to the simultaneous analysis of many specimens and is amenable to automation. These characteristics suggest that this (125)I-labeled staphylococcal protein A immunofiltration technique will be applicable to the rapid identification of other herpesviruses, as well as other clinical isolates.
Effects of time lapse between sputum collection and culturing on isolation of clinically significant fungi
Authors: Hariri A R , Hempel H O , Kimberlin C L , Goodman N L
Published: January 2020

A retrospective study of 34,314 sputum specimens received by a reference laboratory over a 10-year period demonstrated that clinically significant fungi could be isolated even after long periods of delay between collecting and culturing. As a result of this study, it should be stressed that although immediate culturing for fungi is the ideal, specimens should not be rejected because of delays in transport.
Indirect fluorescent-antibody technique for serological diagnosis of La Crosse (California) virus infections
Authors: Beaty B J , Casals J , Brown K L , Gundersen C B , Nelson D , McPherson J T , Thompson W H
Published: January 2020

A clinically relevant indirect fluorescent-antibody technique (IFA) was developed for the serological diagnosis of La Crosse virus infections. The IFA (67%) was as sensitive as the hemagglutination inhibition (58%) and neutralization (58%) tests in the detection of antibodies in acute-phase specimens. Immunoglobulin M antibodies were detected by the IFA test in 48% (11 of 23) of these specimens. Diagnostically significant increases in IFA titer were detected in 86% (19 of 22) of the paired samples. Antibodies were detectable in some patients 7 years after infection; however, the IFA test was not as sensitive as the other two tests in the detection of previous infections.
Use of sodium taurocholate to enhance spore recovery on a medium selective for Clostridium difficile
Authors: Wilson K H , Kennedy M J , Fekety F R
Published: January 2020

Isolation of Clostridium difficile from fecal specimens has been facilitated by the development of a selective and differential medium, cefoxitin-cycloserinefructose agar (CCFA). We substituted 0.1% sodium taurocholate for the 2.5% egg yolk in CCFA and compared the growth of 15 isolates of C. difficile on the resulting medium with growth on conventional CCFA. The taurocholate-containing medium (TCCFA) quantitatively recovered vegetative forms of C. difficile in the same numbers as CCFA medium. Recovery of spores was a mean 1.7 log(10) higher on TCCFA than on CCFA. Thirty-six of 60 patient stool specimens growing C. difficile gave a heavier growth on TCCFA than on CCFA, and 9 failed to yield C. difficile on CCFA. TCCFA detected spores of 75 colony-forming units per ml from artificially inoculated fecal specimens when conventional stool culturing techniques were used. Fluorescence of colonies of C. difficile was more intense on TCCFA than on CCFA. TCCFA was simpler to prepare and, overall, was more sensitive than CCFA.
Superoxol(catalase)test for identification of Neisseria gonorrhoeae
Authors: Saginur R , Clecner B , Portnoy J , Mendelson J
Published: January 2020

The Superoxol (Merck & Co., Inc., Rahway, N.J.) test (catalase test using 30% H2O2) was used to differentiate Neisseria gonorrhoeae from other Neisseria species. A positive test was defined as immediate, brisk bubbling upon dropping 30% H2O2 onto a bacterial colony. One hundred percent of the gonococci were Superoxol positive. Only 1% of Superoxol-positive isolates on Thayer-Martin agar were organisms other than gonococci (99% specificity). The test was more reliable than the coagglutination test. Individual strains of a wide variety of Neisseria and Branhamella species were Superoxol positive. They could usually be differentiated from N. gonorrhoeae by their poor growth on selective media, colonial morphology on nonselective media, and simple biochemical tests. The Superoxol test is an excellent screening test for N. gonorrhoeae. A positive result on a clinical isolate growing on Thayer-Martin agar is strongly suggestive of the presence of gonococci.
Urease testing of mycobacteria with BACTEC radiometric instrumentation
Authors: Damato J J , Collins M T , McClatchy J K
Published: January 2020

A total of 140 mycobacterial isolates from patients treated at Fitzsimons Army Medical Center or the National Jewish Hospital and Research Center and from animal specimens submitted to the National Veterinary Services Laboratory were tested by using a urease procedure modified for use with a BACTEC model 301. Mycobacterial suspensions were prepared by using Middlebrook 7H10 Tween broth. Of the 98 mycobacteria isolates which were urease positive utilizing standard methodology, all were positive using the radiometric procedures. Similarly, all 42 urease-negative isolates were also negative employing the new methodology. Although maximum radiometric readings were observed at 48 h, all positive strains were readily identified 24 h after inoculation without sacrificing either test sensitivity or specificity. Thus, urease testing of mycobacteria, using the modified BACTEC radiometric methodology, was as sensitive, as specific, and more rapid than conventional methods.
Diagnosis of group A streptococcal infections directly from throat secretions
Authors: Edwards E A , Phillips I A , Suiter W C
Published: January 2020

The diagnosis of group A streptococcal disease still relies on isolation of group A streptococcal strains on sheep blood agar followed by presumptive identification based on bacitracin sensitivity or the results of the more precise serogrouping methods such as the Lancefield precipitin test. A technique that would permit rapid identification of streptococcal infections directly from throat secretions would allow immediate appropriate antimicrobial therapy for the management of streptococcal infections to be started. We have been able to identify soluble group A antigen directly from throat secretions by using a latex agglutination test. In a clinical trial in which latex (Streptex group A) and conventional culturing techniques were used, 53 throat secretion cultures were tested: 26 were positive by both procedures, 5 were positive by culture only, 3 were positive by the latex agglutination test only, and 19 were negative by both tests.
Transmission of type B viral hepatitis via eye inoculation of a chimpanzee
Authors: Bond W W , Peterson N J , Favero M S , Ebert J W , Maynard J E
Published: January 2020

To simulate a splash of infectious material in the eyes, plasma from a hepatitis B patient was placed on the corneal surfaces of a chimpanzee. The animal became infected 9 weeks later. This result indicates a need for the use of eye protection in high-risk areas, such as clinical laboratories, hemodialysis units, and dental operatories.
Blood culture cross contamination associated with a radiometric analyzer
Authors: Griffin M R , Miller A D , Davis A C
Published: January 2020

During a 9-day period in August 1980 in a New Jersey hospital, three pairs of consecutively numbered blood cultures from different patients were identified as positive for the same organism (two pairs of Klebsiella pneumoniae and one pair of group A Streptococcus), for each pair, both cultures were positive in the same atmosphere, both organisms had the same sensitivities, and the second of each pair grew at least 2 days after the first and was the only positive blood culture obtained from the patient. When the hospital laboratory discontinued use of its radiometric culture analyzer for 15 days, no more consecutive pairs of positive cultures occurred. Subsequent use of the machine for 9 days with a new power unit but the original circuit boards resulted in one more similar consecutive pair (Staphylococcus epidermidis). After replacement of the entire power unit, there were no further such pairs. Examination of the machine by the manufacturer revealed a defective circuit board which resulted in inadequate needle sterilization. Laboratories which utilize radiometric analyzers should be aware of the potential for cross contamination. Recognition of such events requires alert microbiologists and infection control practitioners and a record system in the bacteriology laboratory designed to identify such clusters.
Sensitivity, specificity, and reproducibility of the automicrobic system (with the Enterobacteriaceae-plus biochemical card) for identifying clinical isolates of Gram- negative bacilli
Authors: Barry A L , Gavan T L , Badal R E , Telenson M J
Published: January 2020

Two independent laboratories tested 1,743 clinical isolates by using the Enterobacteriaceae-plus Biochemical Card in the AutoMicrobic system (AMS) and identical standard reference methods. Included were 55 isolates representing 11 species that cannot be identified by the enterobacteriaceae-plus Biochemical Card computer program; 3 or these isolates were incorrectly identified as Pseudomonas cepacia. With the other 1,688 isolates, 5% of the AMS identifications were considered to be equivocal (probability value, less than 0.80), and the remaining test were 97% accurate (sensitive), Difficulty was observed in the ability of the AMS to identify some H2S-negative Citrobacter freundii species. An AMS response of P. cepacia was also considered nonspecific, because several other organisms were misidentified at P. cepacia. Reproducibility of the system was documented by testing 125 strains on 3-separate days; only 6 strains produced significantly variable results. The AMS (with the Enterobacteriaceae-plus Biochemical Card) was a very satisfactory, automated system for accurately identifying most gram-negative bacilli within 8 to 13 h.
Differential and selective medium for isolation of Yersinia enterocolitica from stools
Authors: Agbonlahor D E , Odugbemi T , Dosunmu-Ogunbi O
Published: January 2020

A new differential and selective medium, DYS agar, was developed and evaluated from the isolation of Yersinia enterocolitica. Ther bile salts content of the medium resulted in high selectivity, and inclusion of arabinose, lysine, and arginine rendered Y. Enterocolitica very distinct from Proteus spp., Pseudomonas spp., and other members of the family Enterobacteriaceae. DYS medium was more efficient for the isolation of Y. enterocolitica from experimentally inoculated fecal specimens than MacConkey, deoxycholate-citrate, and salmonella-shigella agars. Although the medium showed selectivity similar to that of another relatively new medium. Y medium (a selective medium for Y. enterocolitica containing sodium oxalate). DYS agar was found to be superior to Y medium in terms of differentiation of Y. enterocolitica from other intestinal organisms. DYS medium is simple to prepare.
Selective medium for isolation of Actinobacillus actinomycetemcomitans
Authors: Slots J
Published: January 2020

A selective medium, TSBV (tryptic soy-serum-bacitracin-vancomycin) agar, was developed for the isolation of Actinobacillus actinomycetemcomitans, TSBV agar contained (per liter) 40 g of tryptic soy agar, 1 g of yeast extract, 100 ml of horse serum. 75 mg of bacitracin, and 5 mg of vancomycin. The TSBV medium suppressed most oral species and permitted significantly higher recovery of A. actinomycetemcomitans than nonselective blood agar medium. The distinct colonial morphology and positive catalase reaction of A. actinomycetemcomitans easily distinguished this bacterium from Haemophilus aphrophilus, Capnocytophaga species, and a few other contaminating organisms. With the TSBV medium, even modestly equipped laboratories will be able to isolate and identify A. actinomycetemcomitans from clinical specimens.
Detection of infectious mononucleosis heterophil antibody by a rapid, standardized enzyme-linked immunosorbent assay procedure
Authors: Halbert S P , Anken M , Henle W , Golubjatnikov R
Published: January 2020

A rapid, specific, sensitive, standardized, a reproducible enzyme-linked immunosorbent assay (ELISA) procedure has been developed for detecting the heterophil antibody associated with infectious mononucleosis (IM). The IM heterophil antibody used for the solid phase was purified from bovine erythrocyte stroma. The test uses heavy-chain-specific anti-immunoglobulin M (IgM) labeled with alkaline phosphatase and three 10-min incubations. The quantitative results correlated well with horse erythrocyte agglutination titers. Absorption tests confirmed the specificity of the ELISA reactions for IM heterophil antibodies. Neither very high levels of IgM in myeloma sera nor high levels of rheumatoid factor caused false-positive reactions. A number of probable IM cases were encountered which positive by ELISA but negative by the horse erythrocyte slide agglutination test. Absorption studies indicated that these were true-positives for the IM heterophil antibody. The IM heterophil antibodies were confirmed to be predominantly of the IgM class, but moderate proportions of the IgM class were sometimes encountered.
Evaluation of the microenzyme-linked immunosorbent assay with Treponema pallidum antigen
Authors: Pope V , Hunter E F , Feeley J C
Published: January 2020

Whole-cell sonicates of Treponema pallidum, Nichols strain, were evaluated in an enzyme-linked immunosorbent assay (Elisa) for syphilis, and results were read in a Dynatek Microelisa Reader. The antigen was evaluated with sera from patients with syphilis, persons presumed normal, and biological false-positives. Two hundred and ninety-seven sera were tested by the ELISA with T. pallidum antigens, the Venereal Disease Research Laboratory (VDRL) slide test, the fluorescent treponemal antibody absorption (FTA-Abs) test, and the microhemagglutination assay for T. pallidum antibodies (MHA-TP). The results of all of the tests were compared. The ELISA, with 89.3% sensitivity, was less sensitive than the VDRL (93.3%) and FTA-Abs (100.0%) tests but more sensitive than the MHA-TP (76.0%). THe ELISA was considerably more sensitive in primary syphilis than the MHA-TP. Specificity was as follows: ELISA, 98.5%; FTA-Abs test, 97.8%; MHA-TP, 98.2%; and VDRL test, 92.7%. The ELISA has good potential as a confirmatory test in the serodiagnosis of syphilis.
Evaluation of fluorescent antinuclear antibody assays, Crithidia luciliae substrate, and single-stranded DNA-binding capacity in diagnosis of four rheumatic diseases
Authors: Chiang M , Chia D , Barnett E V
Published: January 2020

Sera from groups of patient with systemic lupus erythematosus, mixed connective tissue disease, rheumatoid arthritis, and progressive systemic sclerosis and normal controls were compared, using different antinuclear antibody assays. Hep-II cells, used as a substrate for the detection of antinuclear antibodies, appeared to be more sensitive than rat liver substrate. In addition, the fluorescent patterns were easier to identify on Hep-II cells. All systemic lupus erythematosus sera with antibodies reactive with kinetoplasts of Crithidia luciliae had binding greater than 43% for single-stranded DNA. Based on the high sensitivity of the Hep-II substrate and the relative specificity of high (greater than 43%) binding for single stranded DNA by sera from patients with systemic lupus erythematosus, it appears that these two tests are most useful in differential diagnosis and for the detection of systemic lupus erythematosus.
Pneumonia and bacteremia caused by a previously undescribed Moraxella-like bacterium
Authors: Goetz M B , Jones J
Published: January 2020

Immunocompromised patients are frequently subject to unusual infections. We recently treated a renal allograft recipient for pneumonia due to a hitherto undescribed Moraxella-like bacterium which most closely resembles M-5. M-5 has previously been associated in humans only with dog bites and wound infections. The patient responded well to treatment with aminoglycosides and cephalosporins. Susceptibility to these drugs was demonstrated in vitro by a broth dilution technique. On the basis of the known ability of Moraxella species to colonize the oropharynx and the patient's lack of animal exposure, we propose that our patient's illness was secondary to aspiration of colonized oropharyngeal contents.
Differentiation of herpes simplex virus types 1 and 2 by sensitivity to (E)-5-(2-bromovinyl)-2'-deoxyuridine
Authors: Mayo D R
Published: January 2020

Selective inhibition of herpes simplex virus (HSV) type 1 replication but not of HSV type 2 replication by (E)-5-(2-bromovinyl)2'-deoxyuridine (BVDU) was utilized to differentiate clinical isolates of these viruses. BVDU (0.7 microgram/ml) reduced HSV type 1 PFU by an average of 2.74 log10, but had no effect on the plaque-forming ability of HSV type 2 isolates. The incorporation of BVDU into the media of primary cell cultures used for virus isolation in the clinical laboratory allowed for the correct presumptive identification of HSV types.
Vitamin B6 requirements of nutritionally variant Streptococcus mitior
Authors: Schiller N L , Roberts R B
Published: January 2020

The growth rate of three vitamin B6-dependent Streptococcus mitior (B6DS) and two non-B6DS strains in Todd-Hewitt broth, with and without vitamin B6 supplementation, was examined. Even in optimally supplemented culture media, the growth rate of the three B6DS strains was much slower than that of comparable non-B6DS strains. Uptake studies with [3H] pyridoxine suggest that these B6DS strains cannot assimilate pyridoxine. Although not transported intracellularly, pyridoxine inhibited the growth of B6DS strains in media supplemented with other vitamin B6 analogs, probably by binding to the vitamin B6 transport system and inhibiting the uptake of the other vitamin B6 analogs.
Prevalence of Campylobacter fetus subsp. jejuni in healthy populations in southern India
Authors: Rajan D P , Mathan V I
Published: January 2020

Campylobacter fetus subsp. jejuni was isolated from the feces of 14.8% of a random sample of the healthy population of rural southern India. The rate of isolation was highest in preschool children. This finding emphasizes the need to identify markers of pathogenicity in strains of C. fetus subsp. jejuni, which so far are identified only by cultural characteristics. The pathogenic role of this organism in patients with diarrhoea in tropical developing countries can be understood when such markers of pathogenicity are found.
Purification of genus-specific chlamydial antigen and its separation into several components by ion-exchange chromatography
Authors: Schmeer N , Krauss H
Published: January 2020

Sodium deoxycholate-extracted genus-specific chlamydial antigen was purified from contaminating substances by ion-exchange chromatography with DEAE-sephacel, resulting in a decrease in the complement-fixing activity of the antigen, whereas the enzyme-linked immunosorbent assay activity increased. By successive elution with 0.5, 1.0, and 1.5 M acetate buffer at least three clearly separated components were consistently recovered in 14 trials. The identity of these components as genus-specific chlamydial antigen was demonstrated in enzyme-linked immunosorbent assay tests with specific antisera. The antigenic activity of these components was not diminished by prior treatment of the chlamydial particles with pronase. Antiserum prepared by immunization of rabbits with the antigenic component I of an egg-propagated antigen reacted predominantly with the antigenic components I and II of a cell culture-propagated antigen.
Comparison of guinea pig embryo cells, rabbit kidney cells, and human embryonic lung fibroblast cell strains for isolation of herpes simplex virus
Authors: Landry M L , Mayo D R , Hsiung G D
Published: January 2020

With the recognition of the importance and the prevalence of herpes simplex virus infections and the availability of effective chemotherapy, there has come a demand for rapid, sensitive, and accurate diagnosis. Virus isolation in cell culture remains the most widely employed diagnostic method, and the cell cultures used vary greatly from laboratory to laboratory. Therefore, a study was undertaken to compare several commonly employed human fibroblast cell strains and two nonhuman cells, primary rabbit kidney and guinea pig embryo cells, for the primary isolation of this virus. Our experience in the isolation of herpes simplex virus in the clinical laboratory with these cell systems was evaluated, and reisolations from clinical specimens were performed. We found that guinea pig embryo and rabbit kidney cell cultures were equally sensitive for the isolation of herpes simplex virus from clinical specimens and were superior to the human embryonic lung fibroblast cell strains tested in terms of both sensitivity and rapidity of appearance of cytopathic effect. The use of guinea pig embryo cells for the isolation of this virus from clinical specimens has not been previously reported.
Plesiomonas shigelloides overgrowth in the small intestine
Authors: Penn R G , Giger D K , Knoop F C , Preheim L C
Published: January 2020

An 83-year-old male presented with chronic diarrhea and protein malnutrition associated with Plesiomonas shigelloides overgrowth in the small intestine. This overgrowth was related to achlorhydria and small-bowel diverticula. Tests for heat-labile and heat-stable enterotoxin were negative on the P. shigelloides isolated from both the small bowel aspirate (3 x 10(8) bacteria per ml) and the patient's stool. The patient responded satisfactorily to tetracycline, to which this unusual isolate was susceptible in vitro. The ability to correctly identify and determine the antimicrobial susceptibility of P. shigelloides in the setting of small-bowel overgrowth will help determine appropriate therapy.
Comparison of micro-ID and API 20E in rapid identification of Enterobacteriaceae
Authors: Gooch W M , Hill G A
Published: January 2020

The effectiveness of Micro-ID and API 20E as same-day identification systems for Enterobacteriaceae was evaluated in comparison with conventional identification by using 315 clinical isolates and 90 stock strains. The API 20E system was heavily inoculated according to manufacturer's recommendations for same-day identification. We found that 83 and 81% of isolates provided adequate inocula for Micro-ID and API 20E, respectively, and purity of the heavy inocula was not a problem with either system. Overall agreement with conventional identification at genus and species levels was 93.5% with Micro-ID and 90.2% with API 20E. However, when Klebsiella pneumoniae and K. oxytoca were considered as a single species and Proteus morganii was equated with Morganella morganii, agreement was 95.8 and 90.5%, respectively. Only 83.% of isolates were identified on the day of inoculation by API 20E, in contrast to 94.3% with Micro-ID. The remaining isolates required supplementary overnight testing. Provisional (low selectivity) determinations were constant with conventional identification with 49.3% of isolates with API 20E and 82.6% with Micro-ID. Telephone consultations with the manufacturers to resolve unprinted octal codes required a maximum of 15 min with Micro-ID and from 2 to greater than 48 h with API 20E.
Nutritional and metabolic features of Eubacterium suis
Authors: Wegienek J , Reddy C A
Published: January 2020

We studied the nutritional and metabolic features of Eubacterium suis, an anaerobic animal pathogen that causes cystitis and pyelonephritis in pigs. Peptone-yeast extract-starch (PYS) medium, which contained Trypticase (BBL Microbiology Systems), yeast extract, starch, minerals, cysteine, and sodium carbonate, was shown to support excellent growth of this organism (absorbance at 600 nm = 1.8). Growth was considerably less (absorbance at 600 nm = 0.6) when the starch in the medium was replaced by maltose. Formate, acetate, and ethanol were the major products of fermentation of starch or maltose. The organism appears to require a fermentable carbohydrate for growth since the deletion of starch from PYS resulted in a negligible amount of growth. Growth decreased by approximately 20% when CO2 was rigorously excluded from PYS minus Na2CO3. The deletion of only yeast extract from PYS resulted in a decrease in growth of about 75%, and the simultaneous deletion of both yeast extract and Trypticase resulted in negligible growth. When the yeast extract in PYS was replaced by a defined mixture of purine and pyrimidine bases, vitamins, and amino acids, growth was greater than or equal to 80% that observed in PYS. The deletion of Trypticase from this medium resulted in no detectable growth, suggesting a possible peptide requirement for E. suis growth. Good growth (absorbance at 600 nm = 1.4) was obtained when adenine and uracil were substituted for the mixture of purine and pyrimidine bases in modified PYS; the substitution of pyridoxal, riboflavin, and nicotinic acid for the vitamin mixture gave comparable growth. The nutritional requirement of E. suis apparently reflect the fact that the organism adapts to its natural niche by doing away with certain biosynthetic capabilities which it does not seem to require.
Comparison of three automated systems for antimicrobial susceptibility testing of gram-negative bacilli
Authors: Kelly M T , Latimer J M , Balfour L C
Published: January 2020

Several instruments for automated or semiautomated susceptibility testing are currently available. Three of these instruments, Autobac (General Diagnostics, Warner-Lambert Co., Morris Plains, N.J.), MS-2 (Abbott Laboratories, Dallas, Tex.), and AutoMicrobic system (AMS) (Vitek, Inc., Hazelwood, Mo.) were compared for antimicrobial susceptibility testing of gram-negative bacilli. A total of 207 isolates representing 29 species of gram-negative bacilli were tested simultaneously by each instrument and by a standardized disk diffusion reference method. Nine antimicrobial agents, including ampicillin, carbenicillin, cephalothin, gentamicin, tobramycin, amikacin, tetracycline, trimethoprim-sulfamethoxazole, and nitrofurantoin were tested. Discrepancies between the results of the automated and reference disk diffusion methods were resolved by agar dilution testing. Overall, 93% of the Autobac and MS-2 results and 83% of the AMS results were in agreement with the results obtained by the reference methods. The results of the Autobac, MS-2, and AMS systems respectively included 3.3, 2.3, and 4.2% major and very major discrepancies. Excessive testing discrepancies were found for certain drugs, including ampicillin, tetracycline, and nitrofurantoin, and for certain organisms, including species of Providencia, Serratia, and Citrobacter. The results of this comparison of three automated systems for antimicrobial susceptibility testing indicate that the Autobac and MS-2 instruments provided highly reliable results. The AMS need further development of its susceptibility testing capability to eliminate an unacceptably high number of minor discrepancies.