bluuebunny/crossref_metadata_2025 huggingface dataset:
Pathogenesis of catheter sepsis: a prospective study with quantitative and semiquantitative cultures of catheter hub and segments
Authors: Liñares J , Sitges-Serra A , Garau J , Pérez J L , Martín R
Published: January 2020

Our purpose was to study prospectively the causes, routes of infection, and frequency of catheter-related sepsis in patients on total parenteral nutrition. From January 1981 to January 1984, cultures of 135 subclavian catheters from 135 adult patients were done by quantitative and semiquantitative methods. Twenty patients (14.8%) had catheter-related sepsis. Fourteen episodes (70%) stemmed from an colonized hub. Skin infection (Staphylococcus aureus, 2 cases), total parenteral nutrition mixture contamination (Enterobacter cloacae, 2 cases), and hematogenous seeding of the catheter tip (Yersinia enterocolitica, 1 case, and Streptococcus faecalis, 1 case) accounted for the remaining six septic episodes. The catheter hub is, in our experience, the most common site of origin of organisms causing catheter tip infection and bacteremia.
Measurement of active constitutive beta-D-glucosidase (esculinase) in the presence of sodium desoxycholate
Authors: Edberg S C , Trepeta R W , Kontnick C M , Torres A R
Published: January 2020

The hydrolysis of esculin in the presence of bile has been utilized for many years for the identification of bacteria. It is especially useful in differentiating species of the genus Streptococcus. The procedure is a two-step one. First, the bacterium must grow in a particular concentration of bile, and second, it must hydrolyze esculin. The hydrolysis of esculin has traditionally been determined by the brown-black color that results when one of the hydrolysate products, esculetin, reacts with iron in the medium. The procedure requires incubation for 24 h or more. A method was developed based on the measurement of constitutive beta-glucosidase (esculinase) with the repression of this enzyme by bile equivalent (sodium desoxycholate) that required only 30 min. p-Nitrophenyl-beta-D-glucopyranoside was the esculinase substrate, and sodium desoxycholate was substituted for bile salts. After inoculation, a yellow color was equivalent to the brown-black seen in the 40% bile-esculin reaction. The reagent was dispensed in test tubes and was stable for 6 months. The 30-min procedure correlated well with the conventional 24-h bile-esculin agar tube. Streptococcus pneumoniae could also be identified because of the rapid lysis it exhibited in the substrate solution.
Effects of storage temperature and pH on the stability of eleven beta-lactam antibiotics in MIC trays
Authors: Nickolai D J , Lammel C J , Byford B A , Morris J H , Kaplan E B , Hadley W K , Brooks G F
Published: January 2020

Microdilution MIC test trays containing 11 beta-lactam antibiotics in Mueller-Hinton broth at pH 7.31 or 6.80 were prepared and stored at 4, -10, -25, and -70 degrees C. The drugs tested were ampicillin, ticarcillin, mezlocillin, piperacillin, azlocillin, cefazolin, cefotaxime, moxalactam, cefoperazone, ceftriaxone, and imipenem. MICs for Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa ATCC 27853 were determined at weekly intervals for up to 1 year. The data from the MIC determinations showed the stability of antimicrobial activity over time to be -70 degrees C greater than -25 degrees C approximately 4 degrees C much greater than -10 degrees C. The relative stability at 4 degrees C as compared with that at -10 degrees C cannot be explained by desiccation, as determined by changes in broth sodium concentrations. The relative instability at -10 degrees C may have been caused in part by a temperature fluctuation, resulting in intermittent freezing and thawing of the antibiotics. Some of the drugs appeared to be more stable when diluted in broth at pH 6.80, but endpoints were more difficult to read. Cefazolin and cefoperazone were stable at all four storage temperatures. Cefotaxime, moxalactam, and ceftriaxone also were relatively stable. The other drugs showed moderately rapid to rapid deterioration at each temperature except -70 degrees C. Storage at -25 degrees C is suitable for up to 3 months for many, but not all, beta-lactams; -10 degrees C appears to be unsuitable. Storage at -70 degrees C is recommended.
Rapid detection of bovine herpesvirus type 1 antigens in nasal swab specimens with an antigen capture enzyme-linked immunosorbent assay
Authors: Collins J K , Butcher A C , Teramoto Y A , Winston S
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of bovine herpesvirus type 1 (BHV-1) antigens in nasal swab specimens collected from infected animals. Development of the ELISA involved screening and selection of BHV-1-specific monoclonal antibodies for their ability to capture BHV-1 antigens and for their stability and activity after conjugation to horseradish peroxidase. Forty combinations of capture-conjugate monoclonal antibody pairs were screened for detection of nanogram amounts of purified BHV-1 by using a double-antibody-sandwich ELISA in which antigen and conjugated antibody were simultaneously added to antibody-coated wells. Of the 40 monoclonal antibody pairs, 4 were analyzed further and 1 was selected for routine application to clinical specimens. Of 129 nasal swab specimens collected during the first 10 days after experimental infection with BHV-1, 66 were found to be positive by both virus isolation and ELISA and 34 were positive for infectious virus but negative by ELISA. One specimen was positive by ELISA but negative by virus isolation, and the remaining 28 specimens were negative by both tests. Quantitation of the virus-containing specimens showed that the ELISA had a lower detection limit of 10(3.5) median tissue culture infective doses. The ELISA was judged to be highly useful for diagnosis of BHV-1 infections, since all of the nasal swab specimens that were collected from 12 animals during the first 5 days of the infection, when the clinical signs were the most apparent, were positive.
Cervical osteomyelitis caused by Pseudomonas cepacia in an intravenous-drug abuser
Authors: Smith M A , Trowers N R , Klein R S
Published: January 2020

We report a case of vertebral osteomyelitis caused by Pseudomonas cepacia in a patient with a history of intravenous-drug abuse. P. cepacia infections are usually nosocomial, although community-acquired infections occur more commonly in intravenous-drug abusers than in the general population. Vertebral osteomyelitis caused by P. cepacia has not previously been reported.
Antibody response to individual rubella virus proteins in congenital and other rubella virus infections
Authors: Katow S , Sugiura A
Published: January 2020

Serum samples from patients with various forms of rubella virus infection were tested for antibodies to each of three viral structural proteins by radioimmunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In most sera antibody to E1 protein was the predominant species. Sera from patients with congenital rubella syndrome, however, contained significantly more E2 antibody relative to E1 antibody than did sera from other rubella patients.
Effect of adjuvant vaccines on dengue virus type 2 immune ascitic fluid production
Authors: Gamble W C , George E H , Fears M B , Sanderlin D W , Chappell W A
Published: January 2020

In a comparison of dengue type 2 immune mouse ascitic fluid immunization schedules, a schedule in which adjuvant vaccines were not used produced neutralizing antibody titers that were specific and a mouse mortality rate that was lower, resulting in a greater yield of ascitic fluids. In the schedules in which emulsified adjuvant vaccines were used, the quality of the emulsion had little effect on antibody titer produced.
Problems associated with the use of (E)-5-(2-bromovinyl)-2'-deoxyuridine for typing herpes simplex virus
Authors: Swierkosz E M , Arens M Q , Rivetna K A
Published: January 2020

When 0.5 microgram of (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVdU) per ml was incorporated directly into cell culture medium inoculated with eight known positive specimens, one herpes simplex virus type 1 (HSV-1) isolate grew in the presence of BVdU and was misidentified. By plaque assay, the titers of 15 HSV-1 strains were reduced by more than 3 log10 by BVdU, and the titers of 16 HSV-2 strains were reduced by less than 2 log10. Titers of HSV-1 acyclovir-resistant strains were reduced by less than 1.5 log10, which was characteristic of HSV-2 strains. Thus, typing of HSV isolates in the presence of BVdU by plaque assay is reliable only if information regarding previous antiviral therapy is obtained.
Neisseria meningitidis serogroup 29E (Z') septicemia in a patient with far advanced multiple myeloma (plasma cell leukemia)
Authors: Wachter E , Brown A E , Kiehn T E , Lee B J , Armstrong D
Published: January 2020

A case of septicemia caused by Neisseria meningitidis serogroup 29E (Z') in a patient with plasma cell leukemia is described. The patient developed disseminated intravascular coagulation, had a cardiopulmonary arrest, and died. The effects of altered immune function leading to a predisposition to meningococcal infections are discussed.
Detection of group A streptococcal antigen directly from throat swabs with a ten-minute latex agglutination test
Authors: Miceika B G , Vitous A S , Thompson K D
Published: January 2020

Results obtained with the Culturette brand 10-Minute Group A Strep ID system were compared with culture results to measure the ability of this system to detect group A streptococci directly from more than 800 throat swabs. Our study showed a sensitivity of 92.4% and a specificity of 92.8% for this acid extraction, latex agglutination method when compared with anaerobic culturing for group A streptococci. The results suggest that the 10-Minute Group A Strep ID method may prove to be a useful, very rapid and easy method for diagnosing group A streptococcal pharyngitis directly from throat swabs. Further studies are suggested to determine whether this diagnostic method could stand alone or would have to be used in conjunction with culture.
Determination of the optimum incubation period of blood culture broths for the detection of clinically significant septicemia
Authors: Murray P R
Published: January 2020

The value of incubating blood culture broths for more than 7 days was analyzed. A total of 20,155 blood cultures, consisting of a vented Roche tryptic soy broth (R-TSB) bottle and an unvented Difco thiol broth bottle, was processed; 2,509 organisms were recovered in the R-TSB bottles, and 1,865 organisms in the thiol bottles. Only 32 organisms isolated in the R-TSB bottles and 10 organisms isolated in the thiol bottles were detected after incubation for more than 7 days; 15 of the 32 isolates in the R-TSB bottles and all 10 of the isolates in the thiol bottles were either recovered in other blood cultures or were not considered to be clinically significant. Thus, incubation of the R-TSB bottles and unvented thiol bottles for more than 7 days is unnecessary for the detection of most clinically significant septicemias.
Catheter-associated fungemia caused by Fusarium chlamydosporum in a patient with lymphocytic lymphoma
Authors: Kiehn T E , Nelson P E , Bernard E M , Edwards F F , Koziner B , Armstrong D
Published: January 2020

A case of catheter-associated fungemia caused by Fusarium chlamydosporum is described in a patient with lymphocytic lymphoma. The fungus, which has been isolated from soil but not reported to cause human infection, characteristically produces microconidiophores that are polyphialides bearing microconidia that are spindle-shaped but never globose. Results of in vitro antimicrobial susceptibility tests depended on the test conditions used.
Bacteriuria screening by use of acridine orange-stained smears
Authors: Hoff R G , Newman D E , Staneck J L
Published: January 2020

Acridine orange (AO)-stained smears of 1,042 urine specimens were examined for the presence of bacteria and compared with quantitative culture results. The detection of one or more organisms per three AO fields at X200 magnification was noted in 161 of 162 and 193 of 195 urine specimens that grew greater than 10(5) and 10(4) CFU/ml, respectively, of a clinically relevant organism. However, a high number of false-positive AO smears (356 and 324, respectively) was observed among urines that failed to grow organisms at 10(5) and 10(4) CFU/ml. Sensitivity, specificity, and positive and negative predictive values of AO smears were 99, 58, 26, and 99%, respectively, for cultures of greater than or equal to 10(5) CFU/ml and 98, 59, 32, and 99%, respectively for cultures of greater than or equal to 10(4) CFU/ml. Despite the poor specificity of the AO smear, the very high negative predictive value would allow for the ruling out of bacteriuria defined at 10(4) CFU/ml and would eliminate the need for culture of ca. 50% of the urine specimens in this study. Employment of the rapid, inexpensive AO procedure only as a means to eliminate specimens for culture would allow significant cost savings and permit the laboratory to send out a large number of potentially negative or low-count urine results within a short time of specimen receipt.
Microbiological diagnosis of peritonitis in patients on continuous ambulatory peritoneal dialysis
Authors: Vas S I , Law L
Published: January 2020

The fast and accurate etiological diagnosis of peritonitis in patients on continuous ambulatory peritoneal dialysis is essential. The concentration of larger volumes of peritoneal fluids may yield more isolates than conventional methods. The removal of antibiotics present in the fluids as a consequence of therapy by washing or using antibiotic-removing resins increases the yield considerably. The use of anaerobic culture media is justified if fecal organisms are suspected as the cause of the infection.
Comparative evaluation of the API 20S and AutoMicrobic gram-positive identification systems for non-beta-hemolytic streptococci and aerococci
Authors: Facklam R , Bosley G S , Rhoden D , Franklin A R , Weaver N , Schulman R
Published: January 2020

The API 20S system (Analytab Products, Plainview, N.Y.) and the AutoMicrobic Gram-Positive Identification system (GPI; Vitek Systems, Hazelwood, Mo.) were evaluated for their capacity to identify the non-beta-hemolytic streptococci and aerococci to the species level. The 20S system identified 86% (six of seven strains) of nonhemolytic group B streptococci, whereas 100% of the same group B streptococcal strains were correctly identified by the GPI system. With both systems 99% (134 of 135 strains) of four species of group D enterococcus strains and 92% (24 of 26 strains) of the Aerococcus spp. strains were identified. The 20S system identified 84% (41 of 49 strains) of three species of group D non-enterococcus strains. The GPI system identified 96% of the same group D non-enterococcus strains. The 20S system identified 84% (190 of 226 strains) of 10 species of viridans streptococci; however, supplemental conventional tests were required to identify 49% (110 of the 226 strains) of the viridans strains to the species level. The GPI system identified 79% of the same viridans streptococci without the need for supplemental tests. Both systems identified 84% (161 of 192 strains) of the seven most commonly occurring viridans Streptococcus spp. The 20S system identified 82% (75 of 92 strains) and the GPI system identified 84% (54 of 64 strains) of Streptococcus pneumoniae.
Immune response to bovine herpes herpesvirus type 1 infections: virus-specific antibodies in sera from infected animals
Authors: Collins J K , Butcher A C , Riegel C A
Published: January 2020

The virus specificity of antibodies against bovine herpes virus type 1 was determined with a radioimmunoprecipitation assay and serum collected from natural and experimentally induced infections. By using sequentially collected sera, the development of antibodies to 4 to 5 viral glycoproteins and 11 to 12 nonglycosylated proteins was followed for the first 50 days after infection. The major and most consistent responses in experimentally and naturally infected animals were to four glycoproteins with molecular weights of 102,000, 96,000, 69,000, and 55,000, as well as to a major virion 115,000-molecular-weight nonglycosylated protein. The four glycoproteins were all coprecipitated by a neutralizing monoclonal antibody and were probably involved as target antigens in virus neutralization. Another antigenically unrelated glycoprotein with a molecular weight of 82,000 and a nonglycosylated protein with a molecular weight of 91,000 were also precipitated, but the immune response to these two proteins was transient. Reactivity to gp82 was only weakly detected in serum from naturally infected animals. Contact control animals which did not contract a bovine herpes virus type 1 infection but were exposed to infected animals with signs of severe illness had antibodies which recognized gp102, gp96, gp69 and gp55 as well as p115. These antibodies were present in low amounts and, in contrast to infected animals, did not increase between acute and convalescent sampling.
Thermophilic bacteria: a new cause of human disease
Authors: Rabkin C S , Galaid E I , Hollis D G , Weaver R E , Dees S B , Kai A , Moss C W , Sandhu K K , Broome C V
Published: January 2020

We studied a group of 31 bacterial isolates from clinical specimens, received by the Centers for Disease Control since 1961, which have been denoted thermophilic for their unusual ability to grow at 50 degrees C. Microbiological characteristics were determined for the group, and an assessment of their clinical significance was made based on retrospective chart review. These bacteria are all gram-negative, nonfermentative, nonsporulating rods, most of which grow better at 42 or 50 degrees C than at 35 degrees C. Some of the bacteria could be implicated as the etiological agents for meningitis, endocarditis, and septicemia. Thermophilic bacteria should be considered potential pathogens when isolated from appropriate clinical specimens.
Quantitation of immunoglobulin E antibody to cytomegalovirus by antibody capture enzyme-linked immunosorbent assay
Authors: van Loon A M , van der Logt J T , Heessen F W , van der Veen J
Published: January 2020

An antibody capture enzyme-linked immunosorbent assay was developed for detection of immunoglobulin E antibody to cytomegalovirus (CMV-IgE). Affinity-purified anti-human IgE-coated microtiter plates were used to separate IgE from other classes of antibody in serum. Virus-specific IgE was detected by subsequent incubation with horseradish peroxidase-labeled CMV antigen and substrate. The assay was shown to be very sensitive, since in most positive sera CMV-IgE was still detected at a dilution of 1:5,000. Of 45 patients with primary CMV infection, 43 (96%) were found to produce CMV-IgE. In contrast, CMV-IgE was detected in only 4 (9%) of 44 patients with recurrent CMV infection and in 1 of 144 healthy controls. Furthermore, the level of CMV-IgE in patients with recurrent CMV infection appeared to be lower than that in patients with primary infection. Preliminary examination of successive sera suggested that CMV-IgE is produced somewhat slower than CMV-IgM and -IgA but persists for a shorter period. These results suggest that CMV-IgE may be used as an indicator of primary CMV infection.
Proctitis associated with Neisseria cinerea misidentified as Neisseria gonorrhoeae in a child
Authors: Dossett J H , Appelbaum P C , Knapp J S , Totten P A
Published: January 2020

An 8-year-old boy developed proctitis. Rectal swabs yielded a Neisseria sp. that was repeatedly identified by API (Analytab Products, Plainview, N.Y.), Minitek (BBL Microbiology Systems, Cockeysville, Md.), and Bactec (Johnston Laboratories, Towson, Md.) methods as Neisseria gonorrhoeae. Subsequent testing in a reference laboratory yielded an identification of Neisseria cinerea. It is suggested that identification of a Neisseria sp. isolated from genital or rectal sites in a child be confirmed by additional serological, growth, and antibiotic susceptibility tests and, if necessary, by a reference laboratory. The implications of such misidentifications are discussed.
Detection of bacteriuria and pyuria within two minutes
Authors: Pezzlo M T , Wetkowski M A , Peterson E M , de la Maza L M
Published: January 2020

A study was performed to evaluate two rapid urine screening methods, Bac-T-Screen (Marion Laboratories, Inc., Kansas City, Mo.) and Chemstrip LN (Boehringer Mannheim Diagnostics, BioDynamics, Indianapolis, Ind.), for their ability to screen for bacteriuria and pyuria within 2 min. A total of 1,000 urine specimens were tested with the Bac-T-Screen and the Chemstrip LN and compared with a semiquantitative plate culture method. Of the 1,000 specimens tested, 249 had colony counts of greater than or equal to 10(5) CFU/ml by the culture method. Of these, the Bac-T-Screen detected 94.8% (236 of 249) and the Chemstrip LN detected 84.7% (210 of 249). There were 120 pure cultures of probable pathogens of which the Bac-T-Screen detected 97.5% (117 of 120) and the Chemstrip LN detected 91.7% (110 of 120). Leukocyte counts were performed on all specimens, and both methods have the ability to detect greater than 10 leukocytes per mm3 in a majority (greater than 93%) of the specimens. The cost per test for a negative screen is approximately $1.30 for the Bac-T-Screen and $0.40 for the Chemstrip LN. Overall there is a similar negative predictive value with both methods for bacteriuria and pyuria.
New method for classification of leptospiral isolates belonging to serogroup pomona by restriction endonuclease analysis: serovar kennewicki
Authors: Thiermann A B , Handsaker A L , Moseley S L , Kingscote B
Published: January 2020

The genomes of leptospiral field isolates belonging to serogroup Pomona were analyzed and compared with those of type strains by cleavage with restriction endonucleases. This new classification method shows differences among these organisms not indicated by the conventional serological typing method. No differences were observed among isolates from the United States, Canada, and New Zealand. Although all isolates selected for this study had been serologically typed as belonging to serovar pomona, the restriction endonuclease analysis indicates that they belong to serovar kennewicki. kennewicki, a serovar of North American origin, has recently been eliminated from the official serovar list because it was found to be indistinguishable from serovar pomona by the serological method.
Quantitation of hepatitis B virus (HBV) core antigen in serum in the presence of antibodies to HBV core antigen: comparison with assays of serum HBV DNA, DNA polymerase, and HBV e antigen
Authors: Bredehorst R , von Wulffen H , Granato C
Published: January 2020

A quantitation procedure for hepatitis B core antigen (HBcAg) in serum without prior removal of antibodies to HBcAg is described. The virus nucleoprotein core was released from hepatitis B virus (HBV) particles by treatment with Nonidet P-40 detergent and allowed to form immune complexes with homologous antibodies to HBcAg present in the sera of HBV-infected individuals. After precipitation with 2.0% polyethylene glycol-1.5% Tween 20, the HBcAg immune complexes were dissociated by treatment with 3 M KSCN and then adsorbed onto polystyrene beads in the presence of the SCN- ions. Thereby, HBcAg and antibodies to HBcAg were linked independently of each other to the matrix, and the core antigen could be quantitated directly by incubation of the beads with 125I-labeled anti-HBc. Even in the presence of an excess of antibodies to HBcAg in the polyethylene glycol precipitates, HBcAg could be detected without appreciably affecting the sensitivity. The assay proved to be specific for core determinants and exhibited excellent reproducibility. The application of the HBcAg assay in 185 hepatitis B e antigen-positive sera revealed HBc antigenemia in 99% of the sera containing hepatitis B e antigen at titers of greater than or equal to 1:256 and 43% of the sera with lower hepatitis B e antigen levels. However, only in 6 of the 34 HBcAg-negative sera could HBV DNA be detected by blot hybridization. When correlated with HBV-associated DNA polymerase (DNAP) activity, HBc antigenemia was found in all DNAP-positive sera (n = 95) and in 39% of the hepatitis B e antigen-positive sera without detectable DNAP activity (n = 44). Of the DNAP-negative sera with HBc antigenemia, 94% contained HBV DNA, whereas in the absence of HBcAg, HBV DNA could be detected only in 3 of 27 DNAP-negative sera. With regard to sensitivity, the HBcAg assay appeared to be less sensitive than the hybridization technique, but more sensitive than the DNAP assay.
Studies of metabolites in diarrheal stool specimens containing Shigella species by frequency-pulsed electron capture gas-liquid chromatography
Authors: Brooks J B , Basta M T , el Kholy A M
Published: January 2020

Eleven diarrheal stool specimens and 10 control stool specimens from Cairo, Egypt, were studied by frequency-pulsed electron capture gas-liquid chromatography (FPEC-GLC). Four cases involving Shigella sonnei, three cases involving Shigella boydii, and four cases involving Shigella flexneri were studied. The aqueous stools were centrifuged, extracted with organic solvents, and derivatized to form specific electron-capturing derivatives of carboxylic acids, alcohols, hydroxy acids, and amines. Analyses were performed on high-resolution glass columns with an instrument equipped with an extremely sensitive electron capture detector that is specific for the detection of electron-capturing compounds. The diarrheal stools studied had specific FPEC-GLC profiles and contained metabolic markers that readily distinguished between the Shigella spp. studied and Escherichia coli producing heat-stable or heat-labile enterotoxins. S. sonnei stools contained hexanoic acid, 2-hydroxy-4-methylmethiobutyric acid, and some unidentified alcohols that distinguished this organism from other enteric pathogens. S. boydii produced an acid that was unique for this species, and S. flexneri produced alcohols that could be used to distinguish between it and other enteric organisms. The FPEC-GLC profiles obtained during this study were also very different from those reported earlier for Clostridium difficile and rotavirus. This study presents further evidence that the selectivity and sensitivity of FPEC-GLC techniques can be used to rapidly identify causative agents of diarrhea and detect physiological changes that occur in the gut during the course of diarrheal illness.
Enzyme-linked immunosorbent assay for immunoglobulin G antibody to Pasteurella multocida in rabbits
Authors: Klaassen J M , Bernard B L , DiGiacomo R F
Published: January 2020

Three antigen preparations of Pasteurella multocida, lipopolysaccharide antigen, boiled-cell extract antigen, and boiled whole-bacterium antigen, were used in an enzyme-linked immunosorbent assay (ELISA) to detect rabbit immunoglobulin G antibody to P. multocida. The sensitivity of each antigen preparation was compared by using sera from P. multocida-infected and uninfected rabbits and sera from two rabbits immunized with different serotypes of P. multocida. In the ELISA, all three antigen preparations detected high titers of antibodies in infected rabbits and markedly lower levels in uninfected rabbits. When whole-bacterium or boiled-cell extract antigens were used, the ELISA detected antibodies in sera from both immunized rabbits, but with lipopolysaccharide antigen, only antibody to the homologous serotype was detected. Sera absorbed with P. multocida and Bordetella bronchiseptica, another respiratory pathogen of rabbits, revealed that antibodies detected in the ELISA did not cross-react. Since the lipopolysaccharide antigen was more difficult to prepare and may be type specific, and since the whole-bacterium antigen was the least sensitive, the boiled-cell extract was chosen as the best antigen preparation to use in the ELISA.
Direct appraisal of latex agglutination testing, a convenient alternative to enzyme immunoassay for the detection of rotavirus in childhood gastroenteritis, by comparison of two enzyme immunoassays and two latex tests
Authors: Sambourg M , Goudeau A , Courant C , Pinon G , Denis F
Published: January 2020

During February and March 1984, 207 fecal samples from infants and children with gastroenteritis were tested for rotavirus with four techniques: two enzyme immunoassays (Rotazyme; Abbott Laboratories, North Chicago, Ill., and Enzygnost-Rotavirus; Calbiochem-Behring, La Jolla, Calif.) and two latex agglutination tests (Rotalex; Orion Research, Inc., Cambridge, Mass., and Slidex Rota-Kit; Biomérieux). All stool samples were also tested for yeasts and bacterial pathogens. Electron microscopy was used to investigate discrepant results. We found 47% positive samples with Enzygnost-Rotavirus, 38% with Rotazyme, 37% with Slidex Rota-Kit, and 34% with Rotalex. No specimen was found positive by Rotazyme only or Slidex Rota-Kit only. On the contrary, 12 samples which were positive with Enzygnost-Rotavirus only and 3 which were positive with Rotalex only were not confirmed as positive by electron microscopy. Both enzyme immunoassays gave 6% equivocal results; Slidex Rota-Kit gave significantly fewer equivocal results than did Rotalex: 2.9% versus 9.7% (P less than 0.01). The sensitivity and specificity of latex tests compared favorably with that of enzyme immunoassays. Latex agglutination tests can be performed by unskilled personnel and are rapid and relatively cheap. They appear to be very suitable for routine laboratory work and may prove useful for large-scale screening in developing countries.
Evaluation of the p-nitro-alpha-acetylamino-beta-hydroxypropiophenone differential test for identification of Mycobacterium tuberculosis complex
Authors: Morgan M A , Doerr K A , Hempel H O , Goodman N L , Roberts G D
Published: January 2020

The p-nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) differential test for the identification of Mycobacterium tuberculosis recovered from clinical specimens was evaluated by two laboratories and found to be a rapid and accurate procedure with a specificity exceeding 99%.
A new biotype of Legionella dumoffii
Authors: Edelstein P H , Pryor E P
Published: January 2020

A new biotype of L. dumoffii was isolated from lung and sputum samples of an immunosuppressed patient with pneumonia. This strain differs from other described strains of L. dumoffii in that it fails to produce browning of tyrosine-containing buffered yeast extract medium.
Mycobacterium tuberculosis bacteremia detected by the Isolator lysis-centrifugation blood culture system
Authors: Kiehn T E , Gold J W , Brannon P , Timberger R J , Armstrong D
Published: January 2020

Mycobacterium tuberculosis was detected by the Isolator lysis-centrifugation blood culture system from the blood of a patient with tuberculosis of the breast. The organism also grew on conventional laboratory media inoculated with pleural fluid from the patient.
Isolation of an organism resembling Clostridium barati which produces type F botulinal toxin from an infant with botulism
Authors: Hall J D , McCroskey L M , Pincomb B J , Hatheway C L
Published: January 2020

All reported cases of infant botulism except one have been caused by proteolytic strains (group I) of Clostridium botulinum, toxin types A or B. We describe the cultural and biochemical characteristics of the causative organism of this singular case of infant botulism, caused by type F botulinal toxin. Although this organism produces type F botulinal toxin, it is quite different from proteolytic (group I) C. botulinum, being more closely related to Clostridium barati.
Rapid detection of cytomegalovirus in cell culture by indirect immunoperoxidase staining with monoclonal antibody to an early nuclear antigen
Authors: Swenson P D , Kaplan M H
Published: January 2020

A method for the rapid detection of cytomegalovirus (CMV) in MRC-5 cells 48 h after inoculation with clinical specimens was developed. A commercially available monoclonal antibody to a CMV early nuclear antigen was used in an indirect immunoperoxidase (IPA) staining procedure performed directly on acetone-fixed cell monolayers in standard tubes (16 by 125 mm). Of 190 clinical specimens tested, 30 specimens produced CMV cytopathic effect in tissue culture (TC-CPE) within 14 days after inoculation and, of these 30, 28 were positive for CMV after 48 h by the IPA staining procedure (sensitivity, 93%). Of the remaining 160 clinical specimens negative by TC-CPE within 14 days, 7 were positive by the IPA stain (specificity, 96%). However, three of these seven specimens were positive by TC-CPE upon subculture after the initial 14-day incubation period, and one specimen was overgrown by herpes simplex virus type 2 before CMV cytopathic effect could develop. The mean time to appearance of cytopathic effect for the 30 specimens positive by TC-CPE within 14 days was 6.7 days. These findings indicate that this IPA staining is a useful method for the rapid detection of CMV in cell monolayers inoculated with clinical specimens.
Detection of a Trichosporon beigelii antigen cross-reactive with Cryptococcus neoformans capsular polysaccharide in serum from a patient with disseminated Trichosporon infection
Authors: McManus E J , Jones J M
Published: January 2020

Latex beads coated with anti-Cryptococcus neoformans antibody were agglutinated by serum from a bone marrow transplant recipient having a disseminated infection caused by Trichosporon beigelii. The cryptococcal latex agglutination titer in the serum of the patient rose to 1:2,560 by the time of his death. Necropsy confirmed the disseminated Trichosporon infection and absence of C. neoformans. Cell wall extracts of the isolate of the patient and two additional strains of T. beigelii agglutinated anti-Cryptococcus-coated latex beads. The antigen in the serum of the patient and in the extracts responsible for the agglutination was not destroyed by proteolytic enzymes or heat. A single antigen reactive with rabbit anti-Trichosporon serum could be identified in the serum of the patient and the cell wall extracts by rocket immunoelectrophoresis and crossed immunoelectrophoresis. Rocket immunoelectrophoresis and indirect fluorescent-antibody staining demonstrated that anti-Trichosporon antibody recognized the capsular polysaccharide of C. neoformans.
Identification of mycobacteria by specific precipitation of catalase with absorbed sera
Authors: Wayne L G , Diaz G A
Published: January 2020

Cross-absorbed antisera have been prepared against catalase from reference strains of Mycobacterium asiaticum, Mycobacterium gordonae, Mycobacterium scrofulaceum, Mycobacterium simiae, and Mycobacterium szulgai. A total of 61 strains of mycobacteria were grown in small volumes of liquid medium and disrupted in sealed tubes in a cup horn sonicator, and the extracts were tested by a seroprecipitation technique against each of the reference antibody preparations. All 35 strains that belonged to one of the reference species reacted with the corresponding antibody, and none of the 61 extracts gave a significant cross-reaction with the absorbed antibody to a species to which it did not belong. This method appears to provide a rapid and accurate means of identifying mycobacterial cultures in the diagnostic laboratory.
Detection of adenovirus in nasopharyngeal specimens by radioactive and nonradioactive DNA probes
Authors: Hyypiä T
Published: January 2020

The presence of adenovirus DNA in clinical specimens was analyzed by nucleic acid hybridization assays by both radioactive and enzymatic detection systems. The sensitivity of the hybridization tests was in the range of 10 to 100 pg of homologous adenovirus DNA. Minimal background was noticed with unrelated viral and nonviral DNA. Twenty-four nasopharyngeal mucus aspirate specimens, collected from children with acute respiratory infection, were assayed in the hybridization tests and also by an enzyme immunoassay for adenovirus hexon antigen which was used as a reference test. Sixteen specimens positive by the enzyme immunoassay also were positive in the two nucleic acid hybridization tests, and the remaining eight specimens were negative in all of the tests. The results indicate that nucleid acid hybridization tests with both radioactive and nonradioactive probes can be used for diagnosis of microbial infections.
Epidemiology of rotavirus electropherotypes determined by a simplified diagnostic technique with RNA analysis
Authors: Dolan K T , Twist E M , Horton-Slight P , Forrer C , Bell L M , Plotkin S A , Clark H F
Published: January 2020

The incidence and RNA electropherotypes of rotavirus in stools or rectal swabs of children with diarrhea were studied for three rotavirus seasons (1981 through 1984) in Philadelphia, Pa. We used a simplified RNA analysis method involving polyacrylamide gel electrophoresis followed by silver staining. Phosphate-buffered saline suspensions of the stools and swab eluates were examined directly by polyacrylamide gel electrophoresis-silver staining analysis and enzyme-linked immunoadsorbent assay (Rotazyme; Abbott Laboratories); electron microscopy was performed on solid stool specimens. The RNA analysis results were compared with electron microscopy and enzyme-linked immunosorbent assay results and exhibited a sensitivity and specificity greater than or equal to that of electron microscopy or the enzyme-linked immunosorbent assay. Ten different electropherotypes were detected among the 68 rotavirus RNA-positive specimens examined over the 3-year study. The predominant electropherotype was different in each season. Our results indicate that the polyacrylamide gel electrophoresis-silver nitrate strain RNA analysis of simple unextracted stool suspensions is a uniquely useful diagnostic technique; it rapidly provides both a definitive positive result and immediate determination of the RNA electropherotype, which is of value for epidemiological study.
Investigation by Enzyme-Linked Immunosorbent Assay of
Salmonella
H Antigen-Antibody Reactions
Authors: Bettelheim Karl A. , Maskill William J.
Published: January 2020

An indirect enzyme-linked immunosorbent assay has been used for the detection of
Salmonella
group H antibodies.
Identification of 22 Legionella species and 33 serogroups with the slide agglutination test
Authors: Thacker W L , Plikaytis B B , Wilkinson H W
Published: January 2020

We used the slide agglutination test to determine the serologic relationships of 22 Legionella spp. representing 33 serogroups. Antisera prepared against 14 of the Legionella spp. contained cross-reactive antibodies (1+ or greater) at their working dilutions. Numerous cross-reactions were observed for the blue-white fluorescing Legionella spp. With only three exceptions in the latter group, cross-reactive antibodies were removed by absorption, thereby producing serogroup-specific antisera. For screening tests or for identification only to the genus level, nine polyvalent antiserum pools were prepared. Routine use of slide agglutination test reagents should expand the number of Legionella spp. that can be identified in the clinical laboratory and, at the same time, provide a simpler, less costly test procedure.
Use of rapid screening tests in processing urine specimens by conventional culture and the AutoMicrobic system
Authors: Pfaller M A , Koontz F P
Published: January 2020

Two rapid urine screening tests, the Chemstrip LN (BioDynamics, Indianapolis, Ind.) and the Bac-T-Screen urine screening device (Marion Laboratories, Inc., Kansas City, Mo.), were evaluated as techniques to predict bacteriuria as quantitated by either conventional culture or the AutoMicrobic system (Vitek Systems, Inc., Hazelwood, Mo.). A total of 666 urine specimens were analyzed by both screening tests as well as the AutoMicrobic system and quantitative culture. The sensitivities of both Chemstrip LN and Bac-T-Screen for the detection of low levels of bacteriuria (greater than or equal to 10(3) CFU/ml) were comparable (73.3 and 74.4%, respectively) and were too low to recommend their use as a primary urine screen. Their excellent predictive value of a negative result at the 10(5) CFU/ml level (96 and 97.5%, respectively) makes them potentially useful in predicting urine specimens with less than 10(5) CFU/ml. The use of either of these tests in combination with the AutoMicrobic system markedly decreased the time required to classify urine specimens. Their low cost relative to the AutoMicrobic system urine card makes the use of either test cost effective as a screen for the AutoMicrobic system.
Corynebacterium group D2 as a cause of alkaline-encrusted cystitis: report of four cases and characterization of the organisms
Authors: Soriano F , Ponte C , Santamaria M , Aguado J M , Wilhelmi I , Vela R , Delatte L C
Published: January 2020

In four patients with alkaline-encrusted cystitis, Corynebacterium group D2 was isolated from consecutive urine cultures and stones. Encrusted cystitis occurred in bladders harboring inflammatory or tumorous lesions in patients with chronic or recurrent urinary tract infections appearing after surgery or instrumentation. The urease activity of Corynebacterium group D2 and the neutralization of this enzyme by acetohydroxamic acid are shown. Clinical improvement, disappearance of struvite crystals, and decrease of the urine pH were obtained when these bacteria were eliminated from urine samples. Corynebacterium group D2 strains were highly resistant to many antimicrobial agents but were highly susceptible to norfloxacin and vancomycin when tested at two pHs (7.4 and 8.5).
Comparison of a simple latex agglutination test with hemolysis-in-gel, hemagglutination inhibition, and radioimmunoassay for detection of rubella virus antibodies
Authors: Väänänen P , Häivä V M , Koskela P , Meurman O
Published: January 2020

Rubella virus antibodies were measured in 300 sera from pregnant women visiting a maternity center by using a new, simple latex test, Rubalex. The results were compared with those obtained by using hemolysis in gel, hemagglutination inhibition, and radioimmunoassay. The sensitivity of the latex test was 100, 98.0, and 99.6% when compared with hemolysis in gel, hemagglutination inhibition, and radioimmunoassay, respectively. Its comparative specificity was 96.2, 95.7, and 90.7%, and the predictive value of a positive result was 99.2, 99.2, and 98%, respectively. When assayed with the British standard anti-rubella serum its sensitivity was 11 IU/ml. The latex test gave a positive result within 2 min, and 87% of the positive samples had already reacted after 1 min. The negative results remained as such for at least 8 min. No prozone effect was observed for sera with hemagglutination inhibition titers from 256 to 2,048. We concluded that the latex test, Rubalex, was readily applicable for measuring rubella immunity with a reaction time of 2 min in undiluted samples.
Comparison of serodiagnosis of group B coxsackievirus infections by an immunoglobulin M capture enzyme immunoassay versus microneutralization
Authors: Chan D , Hammond G W
Published: January 2020

A solid-phase enzyme immunoassay (EIA) with the capture antibody principle was developed and evaluated for detection of immunoglobulin M (IgM) specific for group B coxsackieviruses (CBV) in human serum. Of 19 patients who were culture positive for CBV type 4 infections, the EIA for CBV type 4 IgM was more sensitive (84%; 16/19 positive) than a standard microneutralization test (16%; 3/19 positive) and a microneutralization assay on the IgM fractions obtained by sucrose density gradient fractionation of human serum (68%; 13/19 positive). The attempt to produce a serotype-specific EIA was not successful, as IgM antibodies were detected to heterotypic CBV. The pooled CBV types 2 to 5 IgM EIA demonstrated an overall sensitivity of 88% (21/24 positive) in comparison to virus isolation for CBV types 4 and 5. The EIA was specific for IgM to CBV, with the exception of some cross-reactions with hepatitis A IgM-positive sera (15%; 3/20). In comparison to the microneutralization test, the EIA for pooled CBV IgM was more rapid, less cumbersome, and more sensitive. Commercially available reagents used in this study enable the CBV IgM EIA to be developed in diagnostic virology laboratories.
Detection of synergistic hemolytic activity of Staphylococcus aureus with the Cathra replicator
Authors: Boyce J M
Published: January 2020

A replicator method for screening Staphylococcus aureus isolates for synergistic hemolytic activity is described. The technique should also be useful for assessing the hemolytic activity of staphylococci and streptococci on different lots or brands of blood agar because the inoculum and the distances between spot inocula are standardized.
Colistin-oxolinic acid blood agar: a selective medium for the isolation of Gardnerella vaginalis
Authors: Thompson J S
Published: January 2020

Colistin-oxolinic acid medium is proposed as a selective isolation medium for Gardnerella vaginalis. The medium is effective in inhibiting staphylococci and gram-negative bacteria while allowing growth of G. vaginalis.
Detection of endotoxin in the plasma of patients with gram-negative bacterial sepsis by the Limulus amoebocyte lysate assay
Authors: Pearson F C , Dubczak J , Weary M , Bruszer G , Donohue G
Published: January 2020

A total of 120 Limulus amoebocyte lysate (LAL) determinations were made on plasma obtained from normal, healthy human blood donors. Results demonstrated a mean endotoxin level in blood of 0.02 to 1.57 pg/ml. The amount of Escherichia coli endotoxin added to human plasma samples can be quantitated by both nephelometry and turbidimetry. Endotoxin-spiked samples were shown to be significantly different from unspiked samples. When plasma samples were collected from 45 patients hospitalized at three centers, a strong association was demonstrated between a positive Limulus amoebocyte lysate assay and a septic condition. Sensitivity, specificity, and false-positive and false-negative rates for the Limulus amoebocyte lysate assay as a diagnostic test for gram-negative bacteremia were estimated.
Preparation of urine samples for use in commercial latex agglutination tests for bacterial antigens
Authors: Weinberg G A , Storch G A
Published: January 2020

The use of latex agglutination (LA) tests for bacterial antigen detection in urine specimens is hindered by troublesome reactions such as nonspecific agglutination. Therefore, procedures such as boiling or membrane filtration of urine specimens are often used before LA testing. We discovered that the composition of the membrane filter used in filtration has a marked effect on the performance of an LA test used for detection of Haemophilus influenzae type b antigen. False-positive LA reactivity was common in urine specimens from pediatric patients that were processed by membrane filtration through certain filters; furthermore, such reactivity also occurred in LA tests for antigens other than those of H. influenzae. A protein present in urine at low concentrations appeared to be responsible for these phenomena.