bluuebunny/crossref_metadata_2025 huggingface dataset:
Wound infection caused by Kanagawa-negative Vibrio parahaemolyticus
Authors: Johnson D E , Weinberg L , Ciarkowski J , West P , Colwell R R
Published: January 2020

Kanagawa-positive Vibrio parahaemolyticus strains are considered to be human pathogens and are most commonly associated with summer diarrhea. Kanagawa-negative strains are most frequently isolated from the environment and are generally considered to be nonpathogens. We report a wound infection caused by a Kanagawa-negative V. parahaemolyticus strain. The infection occurred in October, an unusual time of the year for a V. parahaemolyticus infection to occur in the mid-Atlantic region of the United States.
Evaluation of a leukocyte dip-stick test used for screening urine cultures
Authors: Sawyer K P , Stone L L
Published: January 2020

A 2-min leukocyte esterase dip-stick test for pyuria was used to screen 409 consecutive clinical specimens received for culture. The leukocyte esterase dip-stick test proved to be a sensitive and inexpensive means of screening urine specimens for the absence of leukocytes and, by inference, for the absence of significant bacteruria.
Association of Pseudomonas maltophilia with malignant lesions
Authors: Nagai T
Published: January 2020

Infections caused by nonfermentative, gram-negative, rod-shaped bacteria (NFGNR) in 564 patients were reviewed. For comparison, 227 patients with Serratia marcescens infections were also studied. Among the patients with NFGNR infections, 24% had some form of malignant solid tumor, and 9% had leukemia and lymphoma. Among the patients with S. marcescens infections, 22% had some type of malignant solid tumor, and 8% had leukemia and lymphoma. The frequency of various species of NFGNR (except Pseudomonas maltophilia and S. marcescens) in patients with a malignancy at the site from which the specimens had been obtained was 5 to 24%. However, 39 of 82 patients (48%) with P. maltophilia infections had a malignancy at the site from which P. maltophilia had been isolated. There were significantly more P. maltophilia infections at the sites of malignant lesions than there were other NFGNR or S. marcescens infections (P less than 0.05).
Simple method for collagenase determination in 38 Pseudomonas aeruginosa strains
Authors: Wellisch G , Cohen E , Cahane Z , Horowitz J
Published: January 2020

Collagenase enzyme activity of 38 Pseudomonas aeruginosa strains and 38 strains of Escherichia coli from various pathological sources was measured by a simple method. This method uses plates with collagen gel. The rate of gel lysis is proportional to the collagenase concentration. The method is simple and requires no special materials or equipment. From the 38 P. aeruginosa strains, 34 were collagenase positive. All 38 strains of E. coli were collagenase negative.
Evaluation of the Directigen Group A Strep test kit
Authors: Miller J M , Phillips H L , Graves R K , Facklam R R
Published: January 2020

The Directigen Group A Strep test kit (Hynson, Wescott, and Dunning, Baltimore, Md.) was tested for its ability to detect group A streptococci directly from 147 throat swabs. The results were compared with results from conventional culture and Lancefield serological grouping tests. The data showed that 121 of 124 culture-negative throat specimens were also Directigen negative (98%) and that 21 of 23 culture-positive specimens were Directigen positive (91%). If specimens that provided less than 10 colonies per plate of beta-hemolytic streptococci were eliminated, all of the culture-positive specimens were Directigen positive. Positive or negative results were available within 65 to 70 min of testing. The Directigen method is relatively simple to perform and easy to interpret and provides accurate assessment of the presence or absence of group A streptococci in throat swabs, with little or no cross-reactivity with other beta-hemolytic groups.
Bacteriology of the teeth from a great white shark: potential medical implications for shark bite victims
Authors: Buck J D , Spotte S , Gadbaw J J
Published: January 2020

Bacteria were cultured for the first time from the teeth of a great white shark (Carcharodon carcharias). Isolates included Vibrio alginolyticus, Vibrio fluvialis, Vibrio parahaemolyticus, and other genera. All are common in the marine environment and some may be associated with wound infections in humans. Shark bite lacerations may serve as a source of these potentially infectious bacteria, particularly Vibrio spp., and should be treated immediately. Antibiotic susceptibility patterns are shown for representatives of Vibrio isolates and indicate that a variety of new agents may be appropriate chemotherapy for shark bite victims.
Production of leukocidin by clinical isolates of Pseudomonas aeruginosa and antileukocidin antibody from sera of patients with diffuse panbronchiolitis
Authors: Homma J Y , Matsuura M , Shibata M , Kazuyama Y , Yamamoto M , Kubota Y , Hirayama T , Kato I
Published: January 2020

The ratio of leukocidin-producing strains to clinical isolates of Pseudomonas aeruginosa was investigated together with the production of protease, elastase, and exotoxin A. We also examined whether these strains contain the common antigen which resides in the cell wall. By using the agar gel diffusion test with specific antisera, we found that 87 of 90 (96.7%) of clinical isolates produced leukocidin. Protease, elastase, and exotoxin A were also produced at high percentages. The common antigen was found to exist in all strains. Next, to estimate antileukocidin antibody in the sera of patients, we used an enzyme-linked immunosorbent assay with horseradish peroxidase-protein A. The sera of 39 patients with diffuse panbronchiolitis (DPB) were investigated for antileukocidin antibody. The mean antileukocidin titer in the sera of 17 DPB patients who were not infected with P. aeruginosa and 5 DPB patients who were transiently infected with the bacteria was about the same as the mean antileukocidin titer in the sera of 11 healthy controls, whereas the mean antileukocidin titer in the sera of 17 DPB patients who were persistently colonized was significantly higher than that in healthy controls. These results indicate that leukocidin was produced at the local site of infection in DPB patients.
Determination of susceptibility of anaerobic bacteria to cefotetan and cefoxitin by the thioglycolate disk elution method
Authors: Barry A L , Packer R R
Published: January 2020

The in vitro activities of two cephamycins, cefotetan and cefoxitin, against 107 anaerobic bacteria were evaluated. The aerobically incubated thioglycolate disk elution technique of Kurzynski et al. (Antimicrob. Agents Chemother. 10:727-732, 1976) was also evaluated to establish the appropriate number of 30-micrograms disks to be added to each tube, assuming that strains with MICs of less than or equal to 16 micrograms/ml are susceptible. Optimal predictive values were obtained when two tubes were prepared for each antimicrobial agent, one with two 30-micrograms disks and the other with three 30-micrograms disks. After 48 h of incubation, resistant strains grew in both tubes and susceptible strains provided no growth or growth less than or equal to 50% of that in the control broth. Growth in one tube but not in the other was considered an equivocal test result indicating the need for additional tests; 5 to 7% of our strains gave equivocal results. Reproducibility studies confirmed that broth microdilution tests were more reproducible than disk elution susceptibility tests.
Criteria for detection of Clostridium difficile toxin production by counterimmunoelectrophoresis
Authors: Rennie R P , Elliott J M , Nardini M A , Thornley J H
Published: January 2020

Counterimmunoelectrophoresis (CIE) was compared with a cytotoxicity assay and isolation of toxin-producing Clostridium difficile for the diagnosis of antibiotic-associated diarrhea. Standardized criteria for CIE positivity were established as two major precipitin lines with specific C. difficile antitoxin. Using these criteria on 425 fecal specimens, we determined the sensitivity of CIE (80%) to be comparable to that of the cytotoxicity assay (82%). Both methods were highly specific. With carefully controlled techniques, CIE can be used in routine diagnostic laboratories to screen for toxigenic C. difficile.
Comparison of lysis-centrifugation with lysis-filtration and a conventional unvented bottle for blood cultures
Authors: Gill V J , Zierdt C H , Wu T C , Stock F , Pizzo P A , MacLowry J D
Published: January 2020

Evaluation of a commercially available lysis-centrifugation blood culture system (Isolator, DuPont Co., Wilmington, Del.) and a lysis-filtration blood culture system for 3,111 cultures showed that both methods had comparable recoveries (73 and 68%, respectively) of significant aerobic and facultatively anaerobic isolates. The unvented conventional blood culture bottle had a recovery rate of 59%. Although the lysis-centrifugation and lysis-filtration systems had comparable recoveries of pathogens, the lysis-centrifugation system had the advantage of having colonies immediately available for further testing. The contamination rate with the lysis-centrifugation system was 3%, compared with 6% with the lysis-filtration system and 0.4% with brain heart infusion.
Diagnosis of postnatally acquired rubella by use of three enzyme-linked immunosorbent assays for specific immunoglobulins G and M and single radial hemolysis for specific immunoglobulin G
Authors: Field P R , Gong C M
Published: January 2020

Three commercial indirect enzyme-linked immunosorbent assays (ELISAs) (RUBELISA, Enzygnost-Rubella, and Rubazyme) and a commercial single radial hemolysis (SRH) test (Rubazone) were evaluated for the diagnosis of acute rubella by testing 41 acute- (less than 7 days postonset) and convalescent-phase (8 to 82 days postonset) serum pairs from cases of rubella previously confirmed by significant change in the hemagglutination inhibition test titer. Specificity was tested by using 10 acute- and convalescent-phase serum samples from patients with rash not confirmed as rubella (control group). In testing for rubella-specific immunoglobulin G (IgG) antibody, Enzygnost-Rubella and Rubazyme confirmed infection in 40 and RUBELISA in 39 of the 41 proven rubella patients. For one patient all three ELISAs failed to show a significant titer rise. No false-positive diagnoses occurred in the control group, although a suspected infection was shown by Rubazyme in one patient. No specific IgM could be detected in this case. Single radial hemolysis confirmed infection in 39 of the 41 proven rubella patients, and one false-positive diagnosis occurred in the control group patients. Of the 43 convalescent-phase serum samples, rubella-specific IgM was detected in 42 by Enzygnost-Rubella, in 41 by RUBELISA M, and in 39 by Rubazyme-M. For a rapid diagnosis with acute-phase sera, specific IgM detection by ELISA was most reliable in hemagglutination inhibition test-positive sera; of 18 such serum samples IgM was shown in 15 by Enzygnost-Rubella, in 13 by RUBELISA M, and in 11 by Rubazyme-M. False-positive specific IgM results were shown by Rubazyme-M in two serum samples from one patient in the control group. These serum samples were negative with the other two ELISA methods.
Characteristics of an unclassified Mycobacterium species isolated from patients with Crohn's disease
Authors: Chiodini R J , Van Kruiningen H J , Merkal R S , Thayer W R , Coutu J A
Published: January 2020

The characteristics of an unclassified Mycobacterium sp. isolated from three patients with Crohn's disease are presented. The organism is extremely fastidious and mycobactin dependent and may require up to 18 months of incubation for primary isolation. Colony morphology is rough. Characteristics are unlike those of any presently defined species. The isolates produced postive niacin, catalase, and 2-week arylsulfatase reactions and were susceptible to neotetrazolium chloride (1:40,000), streptomycin (2 micrograms/ml), and rifampin (0.25 micrograms/ml). Chromogenicity, nitrate reduction, quantitative catalase, Tween hydrolysis, urease, tellurite reduction, pyrazinamidase, and 3-day arylsulfatase tests were negative, and the isolates were resistant to thiophene-2-carboxylic acid hydrazide (10 micrograms/ml) and isoniazid (10 micrograms/ml). Optimum growth in broth was determined to be in 7H9 medium with Dubos oleic albumin complex, Tween 80, and mycobactin J at 37 degrees C without CO2 or agitation and in low medium depth. This Mycobacterium sp. may be a subspecies or biovariant of Mycobacterium paratuberculosis, or it may represent a new species of Mycobacterium. It is suggested that this Mycobacterium sp. may play an etiological role in some cases of Crohn's disease.
Use of trehalose-mannitol-phosphatase agar to differentiate Staphylococcus epidermidis and Staphylococcus saprophyticus from other coagulase-negative staphylococci
Authors: Stevens D L , Jones C
Published: January 2020

Using a plate medium containing trehalose, mannitol, and phenolphthalein diphosphate (TMPA), we differentiated significant clinical isolates of Staphylococcus epidermidis by their lack of acid production in 18 h from other coagulase-negative staphylococci, with our results having a sensitivity (R. S. Galen and S. R. Gambino, Beyond Normality: The Predictive Value and Efficiency of Medical Diagnoses) of 100%, a specificity of 89.9%, and a positive predictive value of 94.8%. With a Taxo A bacitracin disk, which differentiates Staphylococcus species from Micrococcus species, no zone of inhibition was seen for 96% of all staphylococcal strains, with 5 of 26 strains of Staphylococcus saprophyticus exhibiting zone diameters up to 10 mm. By using resistance to a 5-microgram novobiocin disk, we differentiated S. saprophyticus, with our results having a sensitivity of 100%, a specificity of 97.1%, and a positive predictive value of 83.9% on TMPA. These two species represented 77.8% of coagulase-negative staphylococci isolated. Reference strains fo Staphylococcus and Micrococcus species were differentiated by TMPA. The cost of TMPA was compared with that of another method. TMPA was found to offer an inexpensive, sensitive method for rapidly differentiating coagulase-negative Staphylococcus isolates.
Improved detection of bacterial antigens by latex agglutination after rapid extraction from body fluids
Authors: Smith L P , Hunter K W , Hemming V G , Fischer G W
Published: January 2020

Nonspecific agglutination of antibody-coated latex particles, unrelated to the presence of specific bacterial antigens, is a major difficulty with commercial latex particle agglutination tests. Rheumatoid and other factors are known to interfere with latex tests. We studied the use of six chelating, reducing, and anticoagulatory reagents in a rapid extraction of antigen procedure to free heat-stable antigens of Haemophilus influenzae type b and group B streptococcus which had been added to human sera. We also screened sera for the incidence of nonspecific agglutination from the three following groups: 123 patients with positive serology tests, 112 hospitalized patients, and 87 blood donors. The rapid extraction of antigen procedure involved a 1:4 dilution of the sera with each of the six reagents, incubation at 100 degrees C for 3 min, and centrifugation at 13,000 X g for 5 min. Two commercial latex kits were tested (Bactigen and Wellcogen). Nonspecific agglutination was entirely eliminated by each of the six extraction reagents. Sera from 52% of the patients with positive serology tests, 29% of the hospitalized patients, and 28% of the blood donors showed nonspecific agglutination with Bactigen before extraction. Nonspecific agglutination was eliminated in all but one sample after the rapid extraction of antigen procedure. This simple, rapid extraction procedure eliminated nonspecific reactions in cerebrospinal fluids and amniotic fluids and reduced this problem in urines and sera with each commercial kit used on clinical specimens.
Rapid determination of 5-fluorocytosine levels in blood
Authors: Noble M A , Harper B , Grant A G , Bernstein M
Published: January 2020

Levels of 5-fluorocytosine in blood were rapidly calculated by determining the amount of competition in the creatinine iminohydrolase (creatinine deiminase; EC 3.5.4.21) assay for creatinine with the Kodak Ektachem system (Eastman Kodak Co., Rochester, N.Y.). The correlation with bioassay values was extremely high (r = 0.982). Standards and samples were highly stable over time.
Infection of Syrian hamsters with Lyme disease spirochetes
Authors: Johnson R C , Marek N , Kodner C
Published: January 2020

Syrian hamsters were shown to be susceptible to infection by the Lyme disease spirochete Borrelia burgdorferi. Although these spirochetes did not cause any outward signs of illness in these animals, they did cause a generalized infection. Spirochetemia was present during the first 6 days of infection. At 14 days postinfection, spirochetes could be isolated from one or more of the following organs: spleen, eyes, kidneys, liver, testes, and brain. Spirochetes were isolated from the eyes and kidneys of one animal 52 days postinfection, suggesting that these organisms may cause a persistent infection. Virulence of B. burgdorferi is maintained by animal passage but is lost upon prolonged in vitro cultivation.
Characterization of Brucella ovis lipopolysaccharide and its use for diagnosis of ram epididymitis by enzyme-linked immunosorbent assay
Authors: Afzal M , Tengerdy R P , Squire P G , Ellis R P
Published: January 2020

Rough lipopolysaccharide, extracted by a mixture of phenol, chloroform, and petroleum ether from freeze-dried Brucella ovis cells with a yield of 0.71%, contained relatively small amounts of protein and nucleic acid contaminants as compared with lipopolysaccharides from other Brucellae. The crude lipopolysaccharide was suitable as a diagnostic antigen in an enzyme-linked immunosorbent assay for the sensitive and specific detection of ram epididymitis caused by B. ovis infection. In comparative serological tests, the enzyme-linked immunosorbent assay with B. ovis lipopolysaccharide gave better identification of infections and fewer false-negative results than the enzyme-linked immunosorbent assay with sonicated antigen or the complement fixation test.
Evaluation of two bioluminescence-measuring instruments, the Turner Design and Lumac systems, for the rapid screening of urine specimens
Authors: Welch W D , Thompson L , Layman M , Southern P M
Published: January 2020

Two bioluminescence-measuring instruments, the Turner Design and Lumac systems, were compared with a standard plate culture method for their ability to rapidly screen 400 urine specimens. For cultures with less than 1,000 CFU/ml the Turner Design, with old and new evaluation formulas, gave 6.5 and 50.6% false-positive results, respectively, versus 17.6% at greater than or equal to 500 relative light units with the Lumac. For cultures which had greater than 10(5) CFU/ml the Turner Design gave 39% (old formula) and 14% (new formula) false-negative results compared with 4% at less than 200 relative units with the Lumac. The microorganisms most frequently isolated in the false-negative cultures from either system were gram-positive cocci. Predictive values for a positive test at greater than 10(5) CFU/ml were 77.4% (old formula) and 35.7% (new formula) for the Turner Design versus only 50% for the Lumac at greater than or equal to 500 relative light units. Predictive values for a negative test for both instruments were greater than 88% at greater than 10(5) CFU/ml. The Turner Design and Lumac systems were 4.0 and 3.7 times as expensive, respectively, as the plate culture method. Although both systems greatly reduce the time required to process urine specimens, their high costs as compared with that of plate culture, their failure to detect many specimens having greater than 10(5) CFU of gram-positive cocci per ml, and the numerous false-positives reported by both instruments suggest that additional improvements in the systems are warranted.
Detection of beta-glucuronidase in lactose-fermenting members of the family Enterobacteriaceae and its presence in bacterial urine cultures
Authors: Hansen W , Yourassowsky E
Published: January 2020

Four hundred strains of lactose-fermenting Enterobacteriaceae were tested for hydrolysis of p-nitrophenyl-beta-D-glucopyranosiduronic acid, the chromogenic enzyme substrate of beta-glucuronidase. Escherichia coli was found to be homogeneous with respect to beta-glucuronidase: more than 94% of the examined E. coli strains were positive, whereas none of the other lactose-fermenting strains possessed beta-glucuronidase activity. The qualitative beta-glucuronidase test, as rapid and simple as the o-nitrophenyl-beta-D-galactopyranosidase test, proved to be of diagnostic value, especially in the identification of E. coli in primary urine cultures. No significant differences were observed in the results of experiments in which either substrate-impregnated disks prepared in the laboratory or commercially available tablets were used.
Monoclonal antibody-based enzyme-linked immunosorbent assays for identification and serotyping of Vibrio cholerae O1
Authors: Gustafsson B
Published: January 2020

Monoclonal antibodies directed against O-specific antigens of Vibrio cholerae O1 lipopolysaccharide were used in two different enzyme-linked immunosorbent assays (ELISAs), designed for identification and serotyping of V. cholerae O1. In the sandwich ELISA, a monoclonal antibody against the group-specific antigen was used as capture antibody, whereas peroxidase-conjugated monoclonal antibodies directed against group- and type-specific antigens were used as the second antibodies. Monoclonal antibodies were also used in ELISA inhibition tests with whole bacteria as inhibitors in microtiter trays coated with V. cholerae O1 lipopolysaccharide. In addition, the monoclonal antibodies were shown to be useful in slide agglutination tests. The enzyme immunoassays were equally sensitive, showing positive reactions with all V. cholerae O1 strains tested, whereas all V. cholerae non-O1 as well as strains of Escherichia coli, Shigella sonnei, Salmonella spp., Citrobacter freundii, and Brucella abortus were negative. The microtiter application makes the immunoassays suitable with low consumption of reagents for screening of samples from suspected cases as well as from the environment.
Isolation of Wolinella recta and Actinomyces viscosus from an actinomycotic chest wall mass
Authors: Spiegel C A , Telford G
Published: January 2020

In this Note we describe a patient with an actinomycotic chest wall mass from which large numbers of Wolinella recta and Actinomyces viscosus organisms were isolated. This is the first reported association of W. recta with extraoral infection and the tenth report of lung infection by A. viscosus.
Candida lusitaniae: frequency of recovery, colonization, infection, and amphotericin B resistance
Authors: Merz W G
Published: January 2020

Candida lusitaniae recovered from 58 specimens from 13 patients represented less than 1% of the yeasts isolated over a 15-month period. The majority of isolates were recovered from respiratory tract, stool, and urine specimens. Of the 13 patients, 1 had a documented infection associated with septicema. Urine isolates from that patient developed resistance to amphotericin B during therapy.
beta-Lactamase detection in nine staphylococcal species
Authors: Selepak S T , Witebsky F G
Published: January 2020

Detection of beta-lactamase by four different techniques (two acidometric, one chromogenic cephalosporin, and one iodometric) was evaluated with 79 isolates of nine different species of staphylococci. Overall, each technique detected more positive isolates when beta-lactamase production had been induced with oxacillin before testing, and each technique performed equally well when organisms were tested after induction.
Effect of the B-D Urine Culture Kit on an automated bacteriuria screen
Authors: Pezzlo M T , Conway S , Jacobson M , Peterson E M , de la Maza L M
Published: January 2020

The effect of urine collected in the B-D Urine Culture Kit (BDT; Becton, Dickinson & Co., Rutherford, N.J.) on the Autobac urine screen (General Diagnostics, Warner-Lambert Co., Morris Plains, N.J.) was investigated. Upon collection, 1,000 clean-voided urine specimens were divided into sterile urine tubes and BDTs. Within 24 h of collection, urine from each tube was cultured by a semiquantitative plate method and screened by the Autobac system. Overall, when screened by the Autobac system, urine collected in the BDT gave fewer false-positive results: 14.4 compared with 22.7% from the sterile urine tubes. However, in comparison with the sterile urine tubes, the BDT false-negative rate was 10.5 versus 4.7%, the detection time was longer, and the cost was increased.
Oligo-2',5'-adenylate synthetase activity in peripheral blood mononuclear leukocytes in various diseases
Authors: Fujii N , Kotake S , Hirose S , Ohno S , Yasuda I , Sagawa A , Ishikawa K , Minagawa T
Published: January 2020

Interferon induces oligo-2',5'-adenylate synthetase in cells. In various diseases, interferon was detectable in the circulation or was produced spontaneously from peripheral blood mononuclear leukocytes. The oligo-2',5'-adenylate synthetase activity in peripheral blood mononuclear leukocytes was examined in various diseases, including systemic lupus erythematosus, sarcoidosis, Vogt-Koyanagi-Harada disease, and Behcet's disease. The activity of this enzyme was significantly increased in systemic lupus erythematosus (P less than 0.01), sarcoidosis (P less than 0.01), and Vogt-Koyanagi-Harada disease (P less than 0.01) compared with that in controls, but the increase of activity was not significant in Behcet's disease (P greater than 0.05).
Effect of refrigerated storage on cefaclor in Mueller-Hinton agar
Authors: Surprenant A M , Preston D A
Published: January 2020

Cefaclor is less stable than most cephalosporins in media at 35 degrees C. We demonstrated that the activity of cefaclor in Mueller-Hinton agar diminishes continuously at 4 degrees C, resulting in a loss of two-thirds of the activity within 21 days. We recommend that agar dilution plates for this cephalosporin be prepared on the day of their use.
Serological analysis of dermatophyte isolates with monoclonal antibodies produced against Microsporum canis
Authors: Polonelli L , Morace G
Published: January 2020

Hybridoma cells produced by fusing myeloma cells with spleen cells from mice immunized with a soluble antigen of Microsporum canis yielded 30 antibody-producing clones. Six of these clones, propagated as ascites tumors in mice, showed two different types of monoclonal antibodies. The type 1 monoclonal antibody reacted with 17 heterologous and 10 homologous dermatophyte antigens. Type 2 monoclonal antibodies were unable to precipitate three antigens from different isolates of M. canis, thus suggesting the occurrence of different serotypes within the species.
Detection and serotyping of herpes simplex virus in MRC-5 cells by use of centrifugation and monoclonal antibodies 16 h postinoculation
Authors: Gleaves C A , Wilson D J , Wold A D , Smith T F
Published: January 2020

Monoclonal antibodies (Syva Co., Palo Alto, Calif.) were used for the detection and serotyping of herpes simplex virus (HSV) isolates by immunofluorescence 16 h after inoculation of MRC-5 monolayers in 3.7-ml shell vials and after low-speed centrifugation. A total of 119 specimens were inoculated into conventional tube cell cultures and shell vials. Of 98 specimens inoculated on the same day of receipt in the laboratory (fresh specimens), all 23 (23.5%) HSV-positive specimens were identified by serotype in 16 h in shell vials by immunofluorescence, whereas only 8 of 23 HSV-positive specimens (34.8%) produced cytopathic effects in conventional tube cell cultures in this time period. Similarly, of 21 original specimen extracts previously determined to be culture positive for HSV (stored specimens), all were detected and serotyped by the immunofluorescence test with monoclonal antibodies 16 h postinoculation compared with the recognition of only 8 of these isolates (38.1%) by cytopathic effect that soon. This technique of centrifugal inoculation of HSV in shell vials containing MRC-5 cells permitted detection of this virus in all positive specimens with serotype determination within 16 h postinoculation.
Biochemical identification of new species and biogroups of Enterobacteriaceae isolated from clinical specimens
Authors: Farmer J J , Davis B R , Hickman-Brenner F W , McWhorter A , Huntley-Carter G P , Asbury M A , Riddle C , Wathen-Grady H G , Elias C , Fanning G R
Published: January 2020

In 1972 there were only 11 genera and 26 species in the family Enterobacteriaceae. Today there are 22 genera, 69 species, and 29 biogroups or Enteric Groups. This paper is a review of all of the new organisms. It has a series of differential charts to assist in identification and a large chart with the reactions of 98 different organisms for 47 tests often used in identification. A simplified version of this chart gives the most common species and tests most often used for identification. The sources of the new organisms are listed, and their role in human disease is discussed. Fourteen new groups of Enterobacteriaceae are described for the first time. These new groups are biochemically distinct from previously described species, biogroups, and Enteric Groups of Enterobacteriaceae. The new groups are Citrobacter amalonaticus biogroup 1, Klebsiella group 47 (indole positive, ornithine positive), Serratia marcescens biogroup 1, and unclassified Enteric Groups 17, 45, 57, 58, 59, 60, 63, 64, 68, and 69.
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting as a serological tool in the diagnosis of syphilitic infections
Authors: Hensel U , Wellensiek H J , Bhakdi S
Published: January 2020

The utility of sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblotting as a serological tool in the diagnosis of human syphilitic infections was examined. In model experiments, rabbits were immunized with Treponema pallidum or T phagedenis, and the antisera were tested for cross-reactivities with both sets of antigens. A major T. pallidum antigen with a molecular weight of ca. 17,000 appeared to be the most reliable specific antigenic marker as assessed by the immunoblotting technique with peroxidase-labeled second antibodies. Antibodies to this antigen were never detected in hyperimmune rabbit anti-T. phagedenis sera or in the sera of nonsyphilitic humans. In contrast, reactive antibodies were found in all syphilitic human sera and also in liquor samples that were positive in the passive hemagglutination test. Differentiation between immunoglobulin M and immunoglobulin G antibodies was directly possible by applying the respective specific second antibodies. Immunoblotting tests were performed with sera exhibiting low passive hemagglutination test titers and equivocal fluorescent treponemal antibody and rapid plasma reagin card reactions. In more than 60% of these cases, immunoblot positivity with respect to the 17,000-molecular-weight antigen was found. The same results were obtained with partially purified 17,000-molecular-weight antigen. The immunoblot technique should be useful as an additional diagnostic tool for differentiating between true and false-positive serological reactions.
Infections caused by Mycobacterium avium complex in immunocompromised patients: diagnosis by blood culture and fecal examination, antimicrobial susceptibility tests, and morphological and seroagglutination characteristics
Authors: Kiehn T E , Edwards F F , Brannon P , Tsang A Y , Maio M , Gold J W , Whimbey E , Wong B , McClatchy J K , Armstrong D
Published: January 2020

The Mycobacterium avium complex, only rarely described as an invasive pathogen in humans, has recently been reported to frequently cause disseminated disease in patients with the acquired immune deficiency syndrome. Between February 1981 and February 1984 at Memorial Sloan-Kettering Cancer Center, 30 patients with acquired immune deficiency syndrome, 3 patients with leukemia, and 2 patients with congenital severe combined immunodeficiency syndrome developed disseminated M. avium complex infection. Mycobacteria were often found in multiple sites both antemortem and postmortem. Blood cultures were a reliable method for detecting disseminated infection, and the new lysis blood culture systems provided an efficient technique for determining the number of organisms per milliliter of blood. Acid-fast stains and cultures of fecal specimens were also helpful in diagnosing infection. Most of the mycobacteria were serovar 4 (77%), and most (86%) produced a deep yellow pigment. All isolates were susceptible to standard concentrations of clofazimine, cycloserine, and ansamycin, but tended to be resistant to isoniazid, streptomycin, ethambutol, ethionamide, and rifampin.
Simple hemagglutination inhibition test for the diagnosis of toxoplasmosis
Authors: Chang G N , Nemzek J A , Tjostem J L , Gabrielson D A
Published: January 2020

A simple hemagglutination inhibition (HI) test for the serological diagnosis of toxoplasmosis has been developed and evaluated. A total of 84 human and 120 mouse serum samples were tested by the newly developed HI test and compared with an immunoglobulin G-indirect fluorescent antibody test. Statistical analysis of serum titers obtained by using the HI test and the immunoglobulin G-indirect fluorescent antibody test showed a correlation coefficient of 0.89. The diagnostic efficacy of HI when compared with the immunoglobulin G-indirect fluorescent antibody diagnostic test results was 96.43% for human sera and 100% for mouse sera. The unique hemagglutination antigen, derived from Toxoplasma gondii (Rh strain) exotoxin, spontaneously binds with mouse or rat erythrocytes, causing the hemagglutination reaction. In this study, 2, 4, or 8 hemagglutinating units of T. gondii exotoxin was used with Swiss/Webster mouse erythrocytes as an indicator for the HI assay. The results indicate that 8 hemagglutinating units is optimal because this concentration has the least unexplained variability. T. gondii exotoxin was stable for at least 18 months at -70 degrees C. The Toxoplasma HI test we report in this paper is shown to be a fast, easy, highly specific, and sensitive test for the diagnosis of toxoplasmosis.
Effects of products of autolysis of tissue and urine on the viability, morphology, and antigenicity of Leptospira interrogans serovar pomona
Authors: Fairbrother J M
Published: January 2020

The effect of the products of autolysis on Leptospira interrogans serovar pomona was investigated in bovine kidney tissues and urine inoculated with this organism. No viable leptospires were found at 24 h or subsequently after inoculation of kidney tissues or urine. Leptospires and their soluble antigens were rapidly destroyed in tissues stored at 20 degrees C and could not be detected by the fluorescent-antibody test and the enzyme-linked immunosorbent assay. On the other hand, leptospires were detectable by dark-field microscopy and fluorescent-antibody test 38 days after addition to urine. The rate of destruction proceeded less rapidly in tissues stored at 4 degrees C. Under the conditions of this study, soluble leptospiral antigen levels decreased less rapidly than did the number of detectable leptospires. Similarly, breakdown of leptospires, demonstrated by dark-field microscopy and fluorescent-antibody test, progressed during the first 5 days after inoculation of urine, when levels of soluble leptospiral antigen remained constant. Although the number of intact leptospires was markedly reduced after freezing of tissues at -20 degrees C, soluble leptospiral antigen levels remained unaltered. Neither intact leptospires nor their soluble antigens were detected after preservation of tissues in Formalin. However, formolization of urine delayed the destruction of leptospires and increased the levels of soluble leptospiral antigen during at least the first 5 days after inoculation. These results indicate that assays such as the enzyme-linked immunosorbent assay should be useful for the detection of soluble leptospiral antigens in urine and in autolyzing tissues such as those taken from a bovine fetus aborted as a result of leptospiral infection.
Turbidimetric method for quantifying serum inhibition of Limulus amoebocyte lysate
Authors: Novitsky T J , Roslansky P F , Siber G R , Warren H S
Published: January 2020

This study describes a method to quantify the inhibition of lipopolysaccharide (LPS) activity by serum with a turbidimetric Limulus amoebocyte lysate assay. Assays were performed in multiwell microplates, and turbidity was measured as the optical density at 380 nm with a microplate spectrophotometer. LPS potency was measured as the 50% maximal Limulus amoebocyte response (LR50) of LPS diluted with saline. By comparing LR50s in saline, LPSs from various species of bacteria were standardized against the U.S. Reference Standard Endotoxin, lot EC-5. The potency of Escherichia coli O113 and O18 and Serratia marcescens LPSs was found to be equal to that of the reference standard EC-5, whereas LPSs from two salmonella species were half as potent. The least potent LPSs tested, obtained from Klebsiella pneumoniae and E. coli rough mutant J5, were 5- and 10-fold less potent, respectively, than EC-5. As a measure of inhibition, the LR50 of LPS in serum was compared to the LR50 of LPS in saline. Serum inhibited the potency of LPS 103- to 6,400-fold compared with saline. A positive correlation was found between standardized potency in saline and serum inhibition of the various LPSs tested. Thus, LPSs from E. coli O113, O18, and EC-5 and S. marcescens, which exhibited the highest potency in saline, were inhibited the most by serum. Likewise, E. coli J5 and K. pneumoniae LPSs, which were the least potent tested, were the least inhibited. The degree of inhibition of all types of LPS tested increased with increasing serum concentration.
Comparison of standard tube and shell vial cell culture techniques for the detection of cytomegalovirus in clinical specimens
Authors: Gleaves C A , Smith T F , Shuster E A , Pearson G R
Published: January 2020

A monoclonal antibody was used to detect an early antigen of cytomegalovirus (CMV) by fluorescence 16 h after inoculation of MRC-5 monolayers in 1-dram (ca. 3.7-ml) shell vials and low-speed centrifugation. Of 770 specimens (urine, blood, lung tissue, sputum) processed in shell vials, 124 (16%) were positive for the virus at 16 h postinfection. CMV was isolated in standard tube cell cultures (average time, 9 days) from only 88 specimens, but there were no instances (with the exception of 2 blood specimens) in which CMV was recovered from tube cultures but not from shell vials. Additional specimens from 18 patients were positive in the shell vial assay but negative in the conventional tube cell culture assay. Other specimens from 14 of the 18 patients yielded CMV in conventional tube cell cultures. Of the 4 patients from whom CMV was not recovered from other specimens by conventional tube cell culturing, all had evidence of recent CMV infections, as indicated by a fourfold or greater rise in antibody titer. The specificity of the shell vial assay for the detection of CMV is supported by assays of other specimens from the same patients yielding the virus or serological evidence indicating recent infections, the known enhancement of CMV detection after centrifugation of the shell vials, and the distinct and easily recognizable fluorescence confined to the nuclei of CMV-infected cells. Our data indicate that the shell vial cell culture assay for the detection of CMV is as specific as and more sensitive than conventional tube cell culturing for the diagnosis of CMV infections.
Comparison of basal media for culturing Campylobacter jejuni and Campylobacter coli
Authors: Ng L K , Stiles M E , Taylor D E
Published: January 2020

Four strains of Campylobacter jejuni and four strains of Campylobacter coli were used to compare the quantitative growth of Campylobacter cells on blood agar base no. 2 (Oxoid), brucella agar (BBL Microbiology Systems and Difco Laboratories), campylobacter agar base (Difco), Columbia blood agar base (Difco and Oxoid), and Mueller-Hinton agar (Difco and Oxoid). Columbia blood agar base and blood agar base no. 2 were inhibitory to most of the strains tested, as evidenced by reduced (10- to 1,000-fold) colony counts compared with other basal media. One of the brucella agars was inhibitory to two of the C. coli strains. The inhibitory effect of these media could be eliminated by addition of FBP (0.05% each ferrous sulfate hydrate, sodium metabisulfite, and sodium pyruvate) or 7% defibrinated sheep blood. However, addition of FBP or blood to brucella agar, campylobacter agar base, or Mueller-Hinton agar did not significantly affect the count, indicating that supplements are not required in these media for growth of Campylobacter in pure culture.
Radioenzymatic assay for trimethoprim in very small serum samples
Authors: Yogev R , Melick C , Tan-Pong L
Published: January 2020

A modification of the methotrexate radioassay kit (supplied by New England Enzyme Center) enabled determination of trimethoprim levels in 5-microliter serum samples. An excellent correlation between this assay and high-pressure liquid chromatography assay was found. These preliminary results suggest that with this method rapid determination of trimethoprim levels in very small samples (5 to 10 microliters) can be achieved.
Two bacteriophages of Clostridium difficile
Authors: Mahony D E , Bell P D , Easterbrook K B
Published: January 2020

Two temperate bacteriophages of differing morphology and host range were isolated by screening 94 isolates of Clostridium difficile. Phage 41 had a 300-nm flexible tail, whereas phage 56 had a shorter tail with a contractile sheath. Electron microscopy of phage 56 lysates exposed to elevated magnesium concentrations showed small virus-like particles which were 21 nm in diameter. The addition of MgCl2 to semisolid agar overlays enhanced both the titer and plaque size of phage 56. Phage 56 was more temperature labile than phage 41 and demonstrated unusual lability in buffer at pH 7.0. One-step growth and adsorption experiments revealed that both phages had latent periods of about 60 min, but phage 56 adsorbed to its indicator strain more efficiently. Phage 56, which was obtained from a toxigenic strain of C. difficile, was used to lysogenize its nontoxigenic indicator strain, but no conversion to toxigenicity was observed in this strain.
Isolation and detection of multiple yeasts from a single clinical sample by use of Pagano-Levin agar medium
Authors: Yamane N , Saitoh Y
Published: January 2020

A total of 15,234 clinical samples were tested on modified Pagano-Levin agar medium to detect multiple yeast species within a single sample. Samples containing more than one yeast species were estimated to be 8.0% of the total. The most frequent combination of different yeasts was Candida albicans and Torulopsis glabrata.
Lactic acid levels in pleural fluid from patients with bacterial pleuritis
Authors: Riley T V
Published: January 2020

Pleural fluid lactic acid estimations were carried out on 60 samples by gas-liquid chromatography. Lactic acid levels in 12 patients with bacterial pleural infection were statistically significantly higher (mean, 287 mg/dl; range, 135 to 482 mg/dl) than in 18 patients with malignancy (mean, 71 mg/dl; range, 24 to 157 mg/dl) and 30 other patients with pleural effusions (mean, 19 mg/dl; range, 10 to 57 mg/dl). The determination of pleural fluid lactic acid may help in differentiating between empyema and nonbacterial pleural effusions in most cases. It is of particular value when antibiotic therapy has commenced before specimen collection and may be useful for monitoring therapy.
Bacteroides fragilis meningitis
Authors: Odugbemi T , Jatto S A , Afolabi K
Published: January 2020

A fatal case of pyogenic meningitis due to Bacteroides fragilis in a 6-year-old boy is reported. The need for processing cerebrospinal fluid of patients with underlying conditions such as chronic otitis media for recovery of both aerobes and anaerobes is discussed.
Comparison of a latex agglutination test with five other methods for determining the presence of antibody against cytomegalovirus
Authors: Beckwith D G , Halstead D C , Alpaugh K , Schweder A , Blount-Fronefield D A , Toth K
Published: January 2020

A latex agglutination test for determination of antibody against cytomegalovirus was compared with five other methods: a solid-phase fluorescent immunoassay, an indirect hemagglutination test, two solid-phase enzyme immunoassays, and an indirect fluorescent-antibody method, with sera collected from 210 random blood donors. Of the sera tested, 28% were positive for anti-cytomegalovirus by concordance of four or more methods. The latex agglutination test performed well, with a sensitivity of 100%, a specificity of 99%, and positive and negative predictive values of 97 and 100%, respectively. The methods were also evaluated for the number of sera requiring repeat testing, equivocal results after retesting, ease of performance, turnaround time, and technical demands. The tests which best met the requirements for a screening test were the solid-phase fluorescent immunoassay, the indirect hemagglutination test, and the latex agglutination test. The latex agglutination test is a valuable screening tool for detecting total anti-cytomegalovirus which has high sensitivity, high negative predictive value, and rare equivocal results and also has the added advantages of ease of performance and rapid turnaround time.
Porcine rotavirus-like virus (group B rotavirus): characterization and pathogenicity for gnotobiotic pigs
Authors: Theil K W , Saif L J , Moorhead P D , Whitmoyer R E
Published: January 2020

A rotavirus-like virus (RVLV) was isolated from a diarrheic pig from an Ohio swine herd. This virus infected villous enterocytes throughout the small intestine of gnotobiotic pigs and induced an acute, transitory diarrhea. Complete virions were rarely observed in the intestinal contents of infected animals; the predominant particle detected by immune electron microscopy was a corelike particle 52 nm in diameter. The genome of the porcine RVLV was composed of 11 discrete segments of double-stranded RNA that produced an electropherotype distinct from the genome electropherotypes of reovirus, rotavirus, and porcine pararotavirus. Porcine RVLV was antigenically unrelated to rotavirus, porcine pararotavirus, or reovirus but was antigenically related to a bovine RVLV.
Detection of Epstein-Barr virus DNA in mouthwashes by hybridization
Authors: Ambinder R F , Wingard J R , Burns W H , Hayward S D , Saral R , Perry H R , Santos G W , Hayward G S
Published: January 2020

An assay for the presence of Epstein-Barr virus (EBV) DNA was developed by using a cloned EBV DNA probe. After preliminary testing showed the assay to be sensitive and specific, it was applied to 135 mouthwashes from bone marrow transplant recipients, and 21 of these tests were positive. The concentration of EBV DNA in mouthwashes in some cases was as high as 10(8) genome equivalents per ml. When compared with the lymphocyte transformation assay on the same samples, the sensitivity was 75% and the specificity was 97%. In contrast to the lymphocyte transformation assay, the hybridization was semiquantitative and yielded results in 72 h. Potential applications include monitoring the effects of various interventions, such as immunosuppressive and antiviral chemotherapy, on EBV shedding.