bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of a new group of Chlamydia psittaci strains called TWAR
Authors: Kuo C C , Chen H H , Wang S P , Grayston J T
Published: January 2020

A new group of Chlamydia psittaci strains has been identified. They are called TWAR after the laboratory designation of the first two isolates. Twelve strains were isolated from pharyngeal swabs of different persons with acute respiratory disease in Seattle, Wash., during 1983 to 1986. One strain was obtained from the eye of a child during the trachoma vaccine study in Taiwan in 1965. Nine strains were characterized in this study. TWAR organisms formed intracytoplasmic inclusions in HeLa cells which were morphologically typical of C. psittaci and iodine stain negative (contained no glycogen). Immunological analysis with various chlamydia-specific monoclonal antibodies revealed that TWAR strains belong to the genus Chlamydia, are distinct from C. trachomatis, and are serologically unique among C. psittaci. All TWAR strains so far isolated appear identical serologically. TWAR organisms grew poorly in egg and cell cultures and demonstrated low virulence to mice by intracerebral, intranasal, and intravenous inoculation. Available data suggest that the TWAR strain is a primary human pathogen.
Demonstration of immunoglobulin M class antibodies to Toxoplasma gondii antigenic component p35000 by enzyme-linked antigen immunosorbent assay
Authors: Lindenschmidt E G
Published: January 2020

On the basis that 89% of 48 acute-phase toxoplasmosis patients showed immunoglobulin M (IgM) class antibodies to the 35,000-molecular-weight antigenic component (p35000) of Toxoplasma gondii, as demonstrated by IgM immunoblotting, the antigen was purified by sucrose gradient centrifugation and enzyme labeled for use in an enzyme-linked antigen immunosorbent assay (ELA) for the demonstration of IgM class antibodies to the p35000 component. The ELA showed a specificity of 96% with 139 serum specimens at a serum dilution of only 1:5. The test serologically detected 73 symptomatic acute-phase toxoplasmosis patients; 64 were positive in the 19S IgM indirect immunofluorescent-antibody test, and 9 were negative, although they showed IgM antibodies to p35000, as demonstrated by IgM immunoblotting. Also, the ELA turned out to be independent of IgM rheumatoid factors in six acute-phase toxoplasmosis serum specimens.
Toxoplasma antigens recognized by naturally occurring human antibodies
Authors: Potasman I , Araujo F G , Remington J S
Published: January 2020

Sera of most adults have high agglutination test titers to Toxoplasma gondii whether or not the adults have other serological evidence of the infection. This finding has been attributed to the presence of naturally occurring antibodies to T. gondii. Consistent with this observation, we have recently noted that protein blots (PB) of sera of individuals not previously infected with T. gondii had immunoglobulin G (IgG) and IgM antibodies to antigens of the parasite. To further define the antigens recognized by these naturally occurring antibodies, we studied PB of sera of 44 adults and 9 children who had no serological evidence of the infection. Multiple antigens of T. gondii with molecular weights of 15,000 to greater than 205,000 were recognized by IgG and IgM natural antibodies of each of the sera. Although a relatively consistent pattern was noted on the IgM PB of the sera of the adults in the molecular weight range of 48,000 to 85,000, greater heterogeneity was noted on the IgG PB. The most common bands noted on the latter were of approximately 30,000 and 92,000 molecular weight. All of the PB obtained with the serial sera collected at yearly intervals from the children revealed bands; in some cases, new bands had appeared with time, and in others the pattern was constant. In children older than 8 years, the patterns of the PB were similar to those noted in PB of sera of the adults.
Isolation of two strains of Kingella kingae associated with septic arthritis
Authors: Raymond J , Bergeret M , Bargy F , Missenard G
Published: January 2020

Two new cases of infection, a presternal abscess and a spondylodiscitis caused by the recently classified bacterium Kingella kingae, are reported. The main bacteriological characteristics and the susceptibility of the two isolates to antimicrobial agents are described. The pathology of K. kingae, particularly among children, is reviewed.
Effect of holding temperature on isolation of Neisseria gonorrhoeae
Authors: Evans K D , Peterson E M , Curry J I , Greenwood J R , de la Maza L M
Published: January 2020

The effect of holding temperature on the recovery of Neisseria gonorrhoeae was studied. From 300 specimens tested, Thayer-Martin medium plates inoculated and incubated in the presence of CO2 at 35, 22, and 4 degrees C for 24 h before incubation at 35 degrees C yielded 100, 96, and 95% of all isolates ultimately recovered from 82 positive specimens. Although there was a decrease in the quantity of organisms recovered, initial incubation of specimens under refrigeration or at room temperature yielded greater than or equal to 95% of the positive specimens.
Comparative evaluation of New York City and modified Thayer-Martin media for isolation of Neisseria gonorrhoeae
Authors: Greenwood J R , Voss J , Smith R F , Wallace H , Peter C , Nachtigall M , Maier T , Wilber J , Butsumyo A
Published: January 2020

Commercially manufactured New York City (NYC) medium and modified Thayer-Martin (MTM) medium were compared for their ability to isolate Neisseria gonorrhoeae from clinical specimens. Twenty-seven public health laboratories throughout California evaluated 4,802 specimens collected from patients attending either sexually transmitted disease or family planning clinics. Total of 726 and 737 N. gonorrhoeae isolates were recovered from NYC and MTM medium, respectively. Although less contamination was noted on NYC medium, MTM medium was equivalent to commercially prepared NYC medium for the isolation of N. gonorrhoeae from clinical specimens.
Time-resolved fluoroimmunoassay with monoclonal antibodies for rapid diagnosis of influenza infections
Authors: Walls H H , Johansson K H , Harmon M W , Halonen P E , Kendal A P
Published: January 2020

Monoclonal antibodies that are broadly reactive with either influenza A or influenza B viruses were used to develop a 2- to 3-h antigen capture time-resolved fluoroimmunoassay (TR FIA) for detecting influenza viral antigens in both original nasopharyngeal aspirate specimens and in tissue cultures inoculated with nose or throat swab specimens. The lower limit of sensitivity of the assay was about 10 pg of protein as determined with purified influenza A nucleoprotein expressed by recombinant DNA. When the TR FIA was performed with 96 nasopharyngeal aspirate specimens collected during outbreaks of influenza A (H3N2) virus and the results were compared with serodiagnosis results with paired sera, the specificity and sensitivity of TR FIA for the demonstration of influenza A infections were 95 and 85%, respectively. In culture confirmation assays, more than 80% of the swab specimens that grew influenza A or B virus within 7 days could be identified by the TR FIA within 48 h of the inoculation of cells. The results are consistent with those previously reported for respiratory syncytial virus and extend the applicability of monoclonal antibody-based TR FIA for the rapid diagnosis of acute respiratory viral infections.
Dip slide culture of intraoperative peritoneal irrigation fluid for prediction of septic complication in elective colorectal surgery
Authors: Claesson B E , Holmlund D E
Published: January 2020

A controlled prospective study of a simplified technique, the dip slide culture method, for assessment of bacterial concentration in peritoneal irrigation fluid at the end of an elective colorectal operation is presented. The prediction of postoperative surgical infection based on intraoperative culture was compared between this method and a standard streak-plate technique in 190 patients. One gram of metronidazole was given intravenously as prophylaxis on induction of anesthesia and 12 h postoperatively. Intraoperative growth of members of the family Enterobacteriaceae or Staphylococcus aureus was strongly correlated to infection (P less than 0.001). By using this finding as a single criterion for the prediction of sepsis, sensitivity and specificity for the dip slide method were 70.8 and 94.3%, respectively, compared with 79.2 and 94.6%, respectively, for the streak-plate method. Of the 24 infections, 20 (83.3%) were correctly predicted when a combination of the two methods was used. An increasing number of Enterobacteriaceae or S. aureus in the dip slide culture resulted in a steady rise in the rate of infections from 5.7% at 0 CFU to 57% at greater than or equal to 80 CFU compared with 4.1% at 0 CFU/ml and 45.4% at greater than or equal to 800 CFU/ml with the streak-plate method. The differences were statistically significant at the greater than or equal to 5 CFU level with regard to wound infection (P less than 0.001) and deep surgical infection (P less than 0.01). It is concluded that the dip slide is a simple, rapid, and reliable method for the routine assessment of bacterial contamination in colorectal operations.
Age-related rate of seropositivity of antibody to Giardia lamblia in four diverse populations
Authors: Miotti P G , Gilman R H , Santosham M , Ryder R W , Yolken R H
Published: January 2020

We used a solid-phase enzyme immunoassay to determine the age-specific rates of acquisition of antibody to Giardia lamblia in populations living in an inner city area of Baltimore, Md., on an Apache Indian reservation in Arizona, in a rural area of Panama, and in an urban area of Peru (Lima). Antibody to G. lamblia was found in a portion of the adults living in all of the study areas. Similar prevalence rates and quantitative levels of antibody were found in the adults living in Arizona (44%), Panama (48%), and Peru (46%). However, a significantly lower (P less than 0.05) percentage of the adults living in Baltimore (18%) displayed serological evidence of infection. Different patterns of age-associated acquisition of antibody were noted in the study populations. In the United States, children living in Baltimore had low levels of seropositivity throughout childhood, whereas children living on the Arizona Indian reservation showed a progressive acquisition of antibody early in childhood, with adult levels achieved by 8 years of age. In Latin America, children living in Panama attained adult levels of seropositivity between 9 and 20 years of age, whereas children in Peru displayed adult levels of seropositivity in the first 6 months of life. Our findings documented the widespread occurrence of G. lamblia infections in diverse populations. Children living in different areas and under different environmental conditions displayed widely differing rates of acquisition of antibody to G. lamblia, possibly resulting from different levels of sanitation, water contamination, and person-to-person contact. Our studies indicate that quantitative solid-phase immunoassays can be used to study the epidemiology of parasitic infections such as those caused by G. lamblia.
Susceptibility testing of slowly growing mycobacteria by a microdilution MIC method with 7H9 broth
Authors: Wallace R J , Nash D R , Steele L C , Steingrube V
Published: January 2020

Based on previous success with rapidly growing mycobacteria, a microdilution MIC system was devised for slowly growing mycobacterial species using 7H9 broth. Test drugs included isoniazid, rifampin, ethambutol, streptomycin, clofazamine, and sulfamethoxazole. Sixty isolates of four mycobacterial species, including Mycobacterium tuberculosis, from patients who had never received drug therapy were evaluated in the system, as well as 25 drug-resistant isolates and 11 control strains. MICs were read when good macroscopic control growth was evident, a period which varied with each species. Most species exhibited a narrow range of MICs with easily discernible growth endpoints. The aminoglycosides, ethambutol, clofazamine, and sulfamethoxazole were the only drugs with activity against all species at clinically achievable levels in serum. Correlation between susceptibilities by the proportion method in agar with single drug concentrations and the broth method were excellent for M. tuberculosis, M. kansasii, and M. marinum for isoniazid, rifampin, and ethambutol. Isolates of the M. avium complex were much more susceptible in broth than in agar for rifampin, ethambutol, and streptomycin. Given the successful transition of most microbiology laboratories to MIC plates for other bacterial species, this method would allow for testing of multiple drugs at multiple concentrations and has good potential for evaluation of drug combinations and drug-resistant isolates.
Comparison of cerebrospinal fluid C-reactive protein and lactate for diagnosis of meningitis
Authors: Komorowski R A , Farmer S G , Knox K K
Published: January 2020

Levels of C-reactive protein and lactate were determined on 562 consecutive cerebrospinal fluid (CSF) samples from adult patients with a wide variety of central nervous system diseases to compare the sensitivity and specificity of CSF lactate and C-reactive protein for the rapid diagnosis of bacterial meningitis. Neither test alone, together, or in combination with elevated CSF leukocyte count and protein had a predictive value over 60% for a positive test in this group of patients with diverse central nervous system problems. Neither test is useful as a screening test for bacterial or mycotic meningitis. Also, in patients with partially treated bacterial meningitis, the tests are often negative. CSF lactate may be useful in differentiating aseptic from septic meningitis in selected patients.
Pseudomonas maltophilia exoenzyme activity as correlate in pathogenesis of ecthyma gangrenosum
Authors: Bottone E J , Reitano M , Janda J M , Troy K , Cuttner J
Published: January 2020

Ecthyma gangrenosum has not been described during the course of blood stream invasion with Pseudomonas maltophilia, although it occurs with a 30% frequency in Pseudomonas aeruginosa septicemia. We isolated P. maltophilia from the blood and an ecthyma lesion in a leukemic patient. The organism was an avid protease and elastase producer and hence mimicked the exoenzyme profile of invasive P. aeruginosa. The patient responded to moxalactam to which the isolate was susceptible in vitro. On the basis of this report, P. maltophilia may be included among an emerging number of gram-negative bacillary species capable of producing severe cutaneous manifestations of bacteremia.
Passive hemagglutination test for enteric fever
Authors: Petchclai B , Ausavarungnirun R , Manatsathit S
Published: January 2020

A passive hemagglutination (PHA) test for serodiagnosis of enteric fever was developed by sensitizing glutaraldehyde-preserved erythrocytes with lipopolysaccharide from Salmonella serogroups A, B, C, and D singly or simultaneously. The lipopolysaccharide-sensitized erythrocytes were tested with sera from 200 blood donors, 100 patients whose hemoculture was positive for Salmonella species, and 10 patients septicemic for other members of the family Enterobacteriaceae. The PHA test was positive in 90% of 28 acute-phase serum samples from patients with enteric fever from one hospital and in 93% of 72 acute-phase serum samples from another hospital. It was also positive in 100 and 60% of early- and late-convalescent-phase sera, respectively. The PHA test was negative in all patients septicemic for other members of the Enterobacteriaceae. Absorption of sera from patients with enteric fever with lipopolysaccharide from other members of the Enterobacteriaceae did not reduce PHA titers, indicating the specificity of the PHA test. Simultaneous sensitization with lipopolysaccharide from Salmonella serogroups A, B, C, and D was useful as a screening test in a limited trial with 28 acute-phase sera, 10 early-convalescent-phase sera, and 17 late-convalescent-phase sera. The PHA test is indeed a simple, sensitive, specific, and rapid test supplementing hemoculture in laboratory diagnosis of enteric fever.
Effect of repeated in vitro transfer of Pasteurella haemolytica A1 on encapsulation, leukotoxin production, and virulence
Authors: Gentry M J , Confer A W , Craven R C
Published: January 2020

Pasteurella haemolytica serotype 1 was transferred daily for 128 serial passages on both unsupplemented brain heart infusion agar and the same basal medium supplemented with bovine blood, horse serum, and yeast extract. Repeatedly transferred cultures were shown to retain the ability to produce both capsular material and leukotoxin. Furthermore, intact organisms were found to be as toxic in vitro for bovine leukocytes and as virulent for mice as unpassaged cultures. These results indicate that the precaution of using only freshly isolated cultures in the study of this organism may not be necessary.
Cytotoxic necrotizing factor production by hemolytic strains of Escherichia coli causing extraintestinal infections
Authors: Caprioli A , Falbo V , Ruggeri F M , Baldassarri L , Bisicchia R , Ippolito G , Romoli E , Donelli G
Published: January 2020

Two hundred and nineteen strains of Escherichia coli from extraintestinal infections and feces of healthy subjects were examined for hemolysin (Hly) and cytotoxic necrotizing factor (CNF) production and for mannose-resistant hemagglutination. Of 105 strains from extraintestinal infections, 42 (40.0%) were positive for production of both Hly and CNF, and 21 (20.0%) were positive for Hly alone; on the contrary, only 1 Hly- and CNF-positive strain and 2 Hly-positive strains were found among 114 strains from normal stools. CNF production was not found to occur among the nonhemolytic strains, confirming the close association existing between these toxic factors. Hemolytic strains positive for CNF showed mannose-resistant hemagglutination less frequently than did Hly-positive, CNF-negative strains (25.6 versus 82.6%), suggesting the existence of two distinct classes among hemolytic strains of E. coli.
Cross-reaction between Streptococcus pneumoniae and group C streptococcal latex reagent
Authors: Lee P C , Wetherall B L
Published: January 2020

The extent of cross-reactivity between Streptococcus pneumoniae and two commercial streptococcal latex grouping reagents was examined. Cross-reactivity with the group C latex reagent was demonstrated in 91% (20 of 22) of blood cultures tested directly and in 97% (73 of 75) of clinical isolates of various serotypes tested after enzymatic extraction. This is important in the clinical laboratory because it may lead to erroneous identification of organisms present in blood cultures.
Effect of carbohydrates on growth of Ureaplasma urealyticum and Mycoplasma hominis
Authors: Robertson J A , Howard L A
Published: January 2020

We examined the effect of 31 carbohydrates on the growth of Ureaplasma urealyticum and Mycoplasma hominis. Arbutin and its breakdown product, hydroquinone, inhibited growth of both species; the other substrates did not alter the extent of growth. Volatile and nonvolatile end products of carbohydrate metabolism were not detected by gas chromatography.
Comparison of Sabouraud dextrose and Pagano-Levin agar media for detection and isolation of yeasts from oral samples
Authors: Samaranayake L P , MacFarlane T W , Williamson M I
Published: January 2020

The sensitivities of Sabouraud dextrose agar and modified Pagano-Levin agar for the primary isolation of yeasts and the recovery of multiple yeast species from single clinical samples were compared by using oral-rinse samples. Although there was a highly significant positive correlation between the numbers of yeasts recovered from both media, modified Pagano-Levin agar was far superior in detecting multiple yeast species in a single sample. Of 150 oral samples containing yeasts, 23 (15.3%) contained more than one yeast species. The most frequent combination of different yeasts was Candida albicans and Torulopsis glabrata.
Quality control limits for microdilution susceptibility tests with norfloxacin
Authors: Barry A L , Gavan T L , Ayers L W , Fuchs P C , Gerlach E H , Thornsberry C
Published: January 2020

A multilaboratory study was designed to define quality control limits for microdilution susceptibility tests with norfloxacin. The following limits were proposed: for Escherichia coli ATCC 25922, 0.03 to 0.125 micrograms/ml; for Pseudomonas aeruginosa ATCC 27853, 1.0 to 4.0 micrograms/ml; for Staphylococcus aureus ATCC 29213, 0.5 to 2.0 micrograms/ml; and for Streptococcus faecalis ATCC 29212, 2.0 to 8.0 micrograms/ml. The latter represents a change in the previously recommended control limits.
Reactivity of monoclonal antibodies to Rickettsia rickettsii with spotted fever and typhus group rickettsiae
Authors: Anacker R L , Mann R E , Gonzales C
Published: January 2020

Analysis of 15 spotted fever group (SFG) and 2 typhus group strains of rickettsiae with a panel of monoclonal antibodies revealed a number of shared and unique epitopes of the 120- and 155-kilodalton surface proteins. All of the SFG strains but neither of the typhus group strains reacted with antibody to the lipopolysaccharidelike antigen of Rickettsia rickettsii; possibly the lipopolysaccharidelike antigen is the common antigen which defines the SFG. North Carolina and Montana strains of R. rickettsii known to differ slightly in virulence for guinea pigs differed in at least one epitope of the 120-kilodalton protein.
Cryptosporidiosis in patients at a large teaching hospital
Authors: Marshall A R , Al-Jumaili I J , Fenwick G A , Bint A J , Record C O
Published: January 2020

In 1984, all stool samples submitted to a microbiology laboratory (n = 2,197) from a predominantly hospital-based population were examined for Cryptosporidium oocysts after Ziehl-Neelsen staining, but only 0.5% were positive. We suggest that in low-incidence areas, individual laboratories examine stool samples for this organism only in immunosuppressed patients and those in whom symptoms persist.
Comparison of paraffin baiting and conventional culture techniques for isolation of Nocardia asteroides from sputum
Authors: Singh M , Sandhu R S , Randhawa H S
Published: January 2020

Of 1,510 sputum samples examined from 1,016 patients with bronchopulmonary disorders, Nocardia asteroides was isolated from 67 samples by paraffin baiting, as compared with only 30 isolations by the conventional technique of culturing on Sabouraud dextrose agar. This higher efficacy of the paraffin bait technique was found to be statistically significant (P less than 0.001).
Cultivation and characterization of human rotaviruses with "super short" RNA patterns
Authors: Albert M J , Unicomb L E , Bishop R F
Published: January 2020

Two group A, subgroup I, rotavirus strains possessing "super short" RNA patterns were adapted to growth in MA-104 cells. Both produced marked cytopathic effect in primary culture. Reciprocal cross-neutralization titers, polypeptide analysis, and the serum neutralizing antibody response of an infected child suggested that super short viruses are serotypically distinct from the four recognized human serotypes.
Mycobacterium haemophilum infection in a patient with acquired immune deficiency syndrome
Authors: Males B M , West T E , Bartholomew W R
Published: January 2020

Mycobacterium haemophilum was isolated from wrist and ankle aspirates as the organism responsible for tenosynovitis in a patient with acquired immune deficiency syndrome. Mycobacterium isolates recovered from synovial fluid were identified as hemin requiring by their failure to grow on subculture unless the medium was supplemented with hemin. M. haemophilum is of low virulence and rarely associated with infections in humans. This is the first documented case of M. haemophilum infection in a patient with acquired immune deficiency syndrome.
Modified indirect immunofluorescence test for serotyping large numbers of Ureaplasma urealyticum clinical isolates
Authors: Naessens A , Lauwers S
Published: January 2020

A technical modification of the indirect immunofluorescence test for serotyping Ureaplasma urealyticum clinical isolates is described. The use of a tissue culture plate filled with ureaplasma agar made the serotyping easier to perform and proved to be very handy, especially for performance of large series.
Accuracy of single radial hemolysis test for rubella immunity when internal reference standards are used to estimate antibody levels
Authors: Nommensen F E
Published: January 2020

The accuracy and reproducibility of antibody levels obtained by single radial hemolysis with six internal reference sera were evaluated. The test was performed in a clinical laboratory for routine assessment of immunity to rubella infection over a period of 1 year. A linear relationship exists between the antibody titer (expressed in log dilution) and zone diameters. In 43 of 44 test runs the correlation coefficient of the standard curve was over 0.990. Prediction limits of 95% around the curve showed that on replication of the test, zone diameters could be found within less than half a doubling-dilution step. The antibody level can thus be determined more accurately by the single radial hemolysis test than in the conventional hemagglutination inhibition test. This is particularly important in assessing immunity when antibody titers are low, since the hemagglutination inhibition test is less reliable then. The use of standard sera calibrated international units would render results of different laboratories comparable and allow standardization at threshold values.
Identification with monoclonal antibodies of hemolysin produced by clinical isolates of Escherichia coli
Authors: Hugo F , Arvand M , Reichwein J , Mackman N , Holland I B , Bhakdi S
Published: January 2020

Murine monoclonal antibodies were generated against the 107,000-dalton hemolysin encoded by the hemolytic determinant from Escherichia coli LE 2001, and colony blotting was used to assay for production of the hemolysin by 35 hemolytic strains of E. coli and other hemolytic members of the family Enterobacteriaceae of clinical origin. All hemolytic E. coli strains gave positive reactions with two monoclonal antibodies. In contrast, none of the hemolytic, non-E. coli isolates yielded positive colony blots. In addition, Western blotting showed that the hemolysins produced by all clinical E. coli isolates had a similar molecular weight of about 107,000. Discrete antigenic variation may occur in the molecule, since a third monoclonal antibody did not react with the hemolysin from a number of wild-type E. coli strains. Western blot analysis was used to assess the presence of immunoglobulin G (IgG), IgA, and IgM antibodies to E. coli hemolysin in human sera. All 20 of the tested sera from healthy adults contained antibodies to the toxin, with various constellations among the antibody classes. In contrast, sera from five of eight infants aged 8 to 36 months contained no antihemolysin antibodies. We conclude that the 107,000-dalton hemolysin of E. coli is a widespread immunogen that is produced by most or all hemolytic E. coli strains in the human host.
Premarket evaluation of IDS RapID SS/u system for identification of urine isolates
Authors: Halstead D C , Hoffert M R , Colasante G G
Published: January 2020

A total of 170 fresh clinical urine isolates were tested with a premarket configuration of the RapID SS/u system (Innovative Diagnostic Systems, Inc., Atlanta, Ga.), a qualitative micromethod for the identification of selected organisms commonly isolated from urine specimens. Results were compared with those obtained with conventional methods of identifying gram-positive isolates and with the AutoMicrobic system (Vitek Systems, Inc., Hazelwood, Mo.), utilizing Gram-Negative Identification cards for the identification of gram-negative rods. Organisms representing 12 taxa were included in the study. Of the 170 isolates, 163 (95.9%) were correctly identified. A total of 144 strains (84.7%) were correctly identified without additional testing, whereas 19 isolates (11.2%) required further testing. Seven isolates (4.1%) were incorrectly identified. The SS/u system required minimal hands-on time inoculate and interpret reactions. Discrepancies most often occurred with regard to misinterpretation of Escherichia coli and Enterobacter sp. as Citrobacter sp. The IDS RapID SS/u system may indeed prove valuable for the rapid manual identification of urine isolates.
Aspergillus fumigatus-specific antibodies in allergic bronchopulmonary aspergillosis and aspergilloma: evidence for a polyclonal antibody response
Authors: Brummund W , Resnick A , Fink J N , Kurup V P
Published: January 2020

Patients with the Aspergillus-induced diseases allergic bronchopulmonary aspergillosis (ABPA), aspergilloma (fungus ball), and Aspergillus skin test-positive asthma were differentiated immunologically by radioimmunoassay based on their total immunoglobulin E (IgE) and Aspergillus fumigatus-specific IgE levels. In this study, a new, highly sensitive biotin-avidin-linked immunosorbent assay was used to evaluate A. fumigatus-specific antibodies of all immunoglobulin classes. Studied populations included 13 patients with ABPA, 12 with aspergilloma, 9 with Aspergillus skin test-positive asthma, and 9 normal individuals without asthma. A. fumigatus-specific antibodies of all classes were elevated in patients with ABPA, variably elevated in those with aspergilloma, and lowest in the other two groups. This assay demonstrated significantly higher specific IgE antibody levels in the ABPA group over those of the other groups, even with 1:1,000 dilutions of the sera. This study demonstrated that ABPA is a disease characterized by a polyclonal antibody response to Aspergillus antigen and not just a response to IgE and IgG antibody classes. The measurement of other antibody classes, particularly IgD and IgA, could enhance the immunodiagnosis of ABPA. The biotin-avidin-linked immunosorbent assay was found to be a highly sensitive assay that can be a clinically useful alternative to radioimmunoassay in the measurement of A. fumigatus-specific antibodies.
Variations in Western blot banding patterns of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus
Authors: Burke D S , Redfield R R , Putman P , Alexander S S
Published: January 2020

Serum samples from 27 patients infected with human T-cell lymphotropic virus type III (14 with acquired immune deficiency syndrome [AIDS] and 13 with AIDS-related complex) were examined for antibodies to viral proteins by the Western blot method and with four different commercial solid-phase enzyme-linked immunosorbent assays (ELISAs). Virus-specific bands on blots at molecular masses of 64, 55, 53, 41, 31, 24, and 17 kilodaltons were observed. Rank correlation matrices were calculated to relate the intensity of viral bands, stage of illness, and ELISA kit optical densities (ODs). Groups of bands tended to covary in intensity: p17, p24, and p55 (gag gene products); p53 and p64 (pol gene products); and p31 (pol/endonuclease gene product) and p41 (env gene product). Blots of sera from AIDS-related complex patients usually showed strong activity against all viral proteins, while those of sera from AIDS patients characteristically showed strong reactivity only at the pol/endonuclease and env bands. For one ELISA kit (Abbott Laboratories, North Chicago, Ill.), ODs correlated well with the env and pol band intensity scores, while ELISA ODs with other kits (from Litton Industries, Sunnyvale, Calif.; Electro-Nucleonics, Inc., Fairfield, N.J.; and E.I. du Pont de Nemours & Co., Inc., Wilmington, Del.) correlated closely with gag band intensity scores. We conclude that human T-cell lymphotropic virus type III Western blot patterns are determined by (i) viral protein processing pathways and (ii) the stage of illness of the patient and may reflect (iii) the ELISA method used for serum screening.
Detection of hepatitis A virus by extraction of viral RNA and molecular hybridization
Authors: Ticehurst J R , Feinstone S M , Chestnut T , Tassopoulos N C , Popper H , Purcell R H
Published: January 2020

Hepatitis A virus (HAV) RNA was extracted from cell culture, serum, liver, and feces and then detected by molecular hybridization with cloned HAV cDNA. Hybridization was approximately 10-fold more sensitive than immune electron microscopy or radioimmunoassay was and less sensitive than was assays of HAV infectivity in primates or in cell culture. As little as 10(3) 50% infective doses of HAV, or approximately 0.1 pg of viral RNA, was detected by this method. Analysis of fecal specimens from an experimentally infected marmoset and an epidemic of hepatitis A showed that HAV excretion could often be detected later in the illness by hybridization than by radioimmunoassay. This technique should be widely applicable for detection and analysis of HAV RNA.
Diagnosis of chronic Pseudomonas aeruginosa infection in cystic fibrosis by enzyme-linked immunosorbent assay
Authors: Pedersen S S , Espersen F , Høiby N
Published: January 2020

An easily applicable test for diagnosis of chronic Pseudomonas aeruginosa infection in cystic fibrosis by enzyme-linked immunosorbent assay (ELISA) for determination of serum immunoglobulin G to P. aeruginosa was developed. Soluble antigens obtained by ultrasonication of P. aeruginosa, serotypes O:1 to O:17, were used as antigens immobilized to polystyrene microtiter plates. The intraplate, plate-to-plate, and day-to-day variations were 14, 19, and 20%, respectively. Plates coated with the antigens could be stored for at least 64 days at +4 and +22 degrees C without any significant change in activity. Normal values were determined in sera from 164 controls (100 children and 64 adults). The sensitivity and specificity of the ELISA was determined by using serum samples from 243 cystic fibrosis patients and were compared to results with crossed immunoelectrophoresis (CIE). The ELISA could diagnose chronic P. aeruginosa infection with a diagnostic sensitivity of 93% and specificity of 92%. The sensitivity and specificity for the diagnosis of the early stages of chronic P. aeruginosa infection by a single sample were 90 and 100%, respectively, and by using an increased antibody response in paired samples, the sensitivity was 93% and specificity was 87%. There was a statistically significant correlation between antibody levels obtained by ELISA and those obtained by CIE. The sensitivity and specificity of the ELISA were equal to those of CIE, and because of its simplicity, the ELISA is recommended as a routine test in patients with cystic fibrosis.
Cross-reactions between pneumococci and other streptococci due to C polysaccharide and F antigen
Authors: Sørensen U B , Henrichsen J
Published: January 2020

By serological methods, all 83 known types of Streptococcus pneumoniae could be shown to possess C polysaccharide and F antigen. Cross-reactions due to these two antigens between pneumococci and a broad range of most other commonly encountered streptococci were examined. The presence of an antigen closely similar or identical to pneumococcal C polysaccharide was demonstrated in some strains of Streptococcus mitior. Therefore, we conclude that pneumococci cannot be identified serologically from mixed samples without culture. Streptococcal group C antiserum was found to cross-react with pneumococcal F antigen.
Molecular epidemiology of Legionella species by restriction endonuclease and alloenzyme analysis
Authors: Tompkins L S , Troup N J , Woods T , Bibb W , McKinney R M
Published: January 2020

As part of an ongoing investigation into nosocomial Legionella infections at Stanford University Medical Center (SUMC), we applied the technique of restriction endonuclease analysis (REA) to determine strain differences among three species, including Legionella pneumophila, Legionella dumoffii, and Legionella micdadei. A total of 26 human and environmental water isolates from SUMC were selected for REA and compared with control strains that were not epidemiologically linked to SUMC. REA results were compared with results of alloenzyme typing, typing by monoclonal antibodies, and plasmid fingerprinting in all but L. micdadei strains. REA and alloenzyme typing showed that SUMC patient isolates were derived from distinct strains of three species. L. pneumophila strains from SUMC patients were genotypically identical to those isolated from potable water. REA was especially useful in proving that SUMC L. dumoffii patient isolates were derived from a single strain and that patients may have been exposed to a common source(s). REA typing correlated well with alloenzyme typing. These methods complement serologic typing of L. pneumophila and provide discriminating capability between strains of other Legionella species such as L. dumoffii, for which serologic types have not been identified. In addition, REA typing is somewhat easier to perform than alloenzyme typing and can be done in clinical laboratories.
Comparison of a novel trimethoprim-sulfamethoxazole-containing medium (XT80) with kanamycin agar for isolation of antibiotic-resistant organisms from stool and rectal cultures of marrow transplant patients
Authors: Hamilton D J , Ulness B K , Baugher L K , Counts G W
Published: January 2020

A new medium (XT80) containing trimethoprim-sulfamethoxazole (TMP-SMZ) was characterized and compared with kanamycin-containing tryptic soy agar (KA) for the recovery of multiply resistant organisms (MRO) in rectal and stool cultures. Cultures from 151 patients hospitalized for bone marrow transplantation were screened for MRO. A total of 366 MRO were recovered from 702 cultures on 94 patients during a 6-month period. XT80 detected more gram-negative bacilli and Corynebacterium spp. than KA. Detection of Staphylococcus spp. was equivalent for the two media. Multiple-antibiotic resistance, defined as resistance to three or more classes of antibiotics, was confirmed by standard agar disk diffusion susceptibility testing. Growth on XT80 correctly identified heteroresistant strains of methicillin-resistant Staphylococcus spp. XT80 more rapidly detected thymidine-dependent mutants of Staphylococcus spp. and members of the family Enterobacteriaceae. Lipophilic Corynebacterium spp., including Corynebacterium group JK, also were more readily detected with XT80. TMP-SMZ given as prophylaxis against Pneumocystis carinii infection exerts a selective pressure on organisms that colonize immunocompromised patients and appears to select for colonization with MRO. Colonization with MRO preceded infection for 94% of 36 patients who developed bacteremia. XT80 is a useful screening tool; growth on this medium correlates closely with resistance to TMP-SMZ and is as accurate a predictor as KA for the carriage of MRO.
Reliability of high-content disks and modified broth dilution tests for detecting staphylococcal resistance to the penicillinase-resistant penicillins
Authors: Barry A L , Jones R N
Published: January 2020

In vitro susceptibility tests were performed with 271 isolates of Staphylococcus species (204 Staphylococcus aureus), including 110 strains resistant to the penicillinase-resistant penicillins. Disks containing 5 or 10 micrograms of methicillin, 1 or 4 micrograms of oxacillin, and 1 or 4 micrograms of nafcillin were evaluated. After a full 24 h of incubation at 35 degrees C, tests with 1-microgram oxacillin disks provided optimal results. Use of the more potent oxacillin, nafcillin, or methicillin disks only increased the number of false-susceptible test results. For broth microdilution tests, 2% NaCl should be added to cation-supplemented Mueller-Hinton broth, and MICs should be recorded after a full 24 h at 35 degrees C. Microdilution tests with oxacillin in broth with 2% NaCl were more reliable than similar tests with methicillin.
Optimal dilution susceptibility testing conditions, recommendations for MIC interpretation, and quality control guidelines for the ampicillin-sulbactam combination
Authors: Jones R N , Barry A L
Published: January 2020

The ampicillin-sulbactam combination was evaluated in vitro to determine the optimal susceptibility testing conditions among five combination ratios and four fixed concentrations of sulbactam. The organisms tested were markedly resistant to aminopenicillins and most other beta-lactams. The ratio of 2:1 is recommended to assure recognition of the ampicillin-sulbactam spectrum and minimize false-susceptible results among strains known to be resistant to this combination. Proposed MIC breakpoint concentrations were compatible with levels in serum achieved with recommended clinical doses. Cross-resistance analyses comparing ampicillin-sulbactam and amoxicillin-clavulanate showed comparable activity and spectra. However, the major interpretive disagreement was sufficient to require separate testing of these aminopenicillin-inhibitor combinations. The recommended ampicillin-sulbactam MIC susceptibility breakpoints are as follows: (i) less than or equal to 8.0/4.0 micrograms/ml for tests against members of the family Enterobacteriaceae, anaerobes, nonenteric gram-negative bacilli, staphylococci, Haemophilus influenzae, and Branhamella catarrhalis; (ii) the ampicillin MICs alone interpreted by National Committee for Clinical Laboratory Standards criteria should predict ampicillin-sulbactam susceptibility for the enterococci, streptococci, and Listeria monocytogenes. MIC quality control ranges were determined by multiple laboratory broth microdilution trials for the ampicillin-sulbactam 1:1 and 2:1 ratio tests.
Detection of Gardnerella vaginalis in vaginal specimens by direct immunofluorescence
Authors: Hansen W , Vray B , Miller K , Crokaert F , Yourassowsky E
Published: January 2020

The preparation of a fluorescein-labeled Gardnerella vaginalis polyclonal antibody is described, and its usefulness is assessed for the detection of this microorganism in vaginal samples obtained from 263 women attending the gynecological department of a general hospital, 66 of whom harbored an intrauterine device. The direct immunofluorescence technique was positive for G. vaginalis in 21% of the specimens, whereas only 12.5% of the total bacteriological cultures were positive. The frequency was higher in patients harboring intrauterine devices since 34.8% exhibited positive immunofluorescence and 30.3% exhibited positive cultures.
Isolation of Staphylococcus aureus L forms from experimentally induced bovine mastitis
Authors: Owens W E
Published: January 2020

Bacterial L forms were isolated from milk samples of dairy cattle infected experimentally with Staphylococcus aureus. Initially, bacterial L forms were induced in vitro from 12 of 44 S. aureus strains isolated from bovine mastitis. Cows were experimentally infected in two experiments with strains shown in vitro to be easily inducible to L form and with S. aureus Newbould 305. Each quarter of the mammary gland was infected with either 10(3) or 10(6) CFU of the test strains. Treatment was initiated with 100,000 U of penicillin G per quarter at the first signs of clinical mastitis. Milk samples were collected daily and cultured on bovine blood agar and PPLO agar (Difco Laboratories, Detroit, Mich.) with 10% horse serum and 5% NaCl. Staphylococcal L forms were isolated from milk samples collected from infected glands in both experiments after antibiotic therapy. Glands with the highest concentrations of leukocytes and bacteria were most likely to yield L forms in milk samples after treatment was initiated. Cows harboring L forms typically yielded parental organisms after cessation of antibiotic therapy. No detectable changes occurred in antibiotic susceptibilities, coagulase production, or biochemical activities in strains induced to L form followed by reversion to the parental form. These results demonstrated that L forms can occur during treatment of bovine mastitis and that L forms may be one explanation for the poor response of staphylococcal bovine mastitis to antibiotic therapy.
Comparison of different procedures, transport media, and enrichment media for isolation of Campylobacter species from healthy laying hens and humans with diarrhea
Authors: Sjögren E , Lindblom G B , Kaijser B
Published: January 2020

A comparison was made with some of the available transport and enrichment media for the isolation of Campylobacter species. The merits of duplicate versus single sampling were also investigated. Fecal samples from 721 healthy laying hens and 328 patients with diarrhea were included in the study. The media compared were modified Stuart medium, Campythio-glycolate medium, semisolid motility test medium, and Campylobacter enrichment broth. The isolation rate of Campylobacter species was significantly increased by taking duplicate instead of single samples. The highest outcome was found when one of the two samples was an enrichment medium, such as semisolid motility test medium.
Comparison of serologic screening tests for brucellosis
Authors: Moyer N P , Evins G M , Pigott N E , Hudson J D , Farshy C E , Feeley J C , Hausler W J
Published: January 2020

The slide agglutination test (SAT), microagglutination test (MAT), and card agglutination test (CAT) were compared with each other, using the tube agglutination test (TAT) as the standard method, by two reference laboratories to determine effectiveness as screening tests for human brucellosis. TAT titers of 1,253 sera tested in both laboratories were compared. In one laboratory, 1,270 sera were tested by the TAT and SAT, while the other laboratory tested 1,261 sera by both methods. Of these sera, 1,155 were tested in one laboratory by the CAT and 187 sera were tested by the MAT. Compared with that of the TAT (greater than or equal to 160 positive), the sensitivities were 97 to 100% (SAT), 90% (CAT), and 88% (MAT). The specificities were 88 to 89% (SAT), 98% (CAT), and 88% (MAT). For populations with a low prevalence of disease, increased specificity offers higher predictive value, so the CAT and MAT are preferable for screening purposes and the choice between tests depends on the number and frequency of tests performed. All sera reactive in the CAT and MAT should be retested with the TAT.
Improved selective culture media for Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus
Authors: Holm A , Rabe P , Kalfas S , Edwardsson S
Published: January 2020

By modifying the previously described media tryptic soy-serum-bacitracin-vancomycin (TSBV) agar and tryptic soy-serum-bacitracin-vancomycin-fluoride (TSBVF) agar, two improved selective culture media were developed for isolation and enumeration of Actinobacillus actinomycetemcomitans (A medium) and Haemophilus aphrophilus (H medium) in oral specimens. Both media were supplemented with fusidic acid and spiramycin, and carbenicillin was also added to A medium. The growth yields of pure cultures of A. actinomycetemcomitans on A medium and of H. aphrophilus on H medium were comparable with those on the reference media. Compared with blood agar, the selective media inhibited these species about 10-fold or less. In addition, A and H media suppressed the growth of pure cultures of Capnocytophaga spp. and Neisseria spp., commonly found as contaminants on TSBV and TSBVF, 10(5) times or more compared with that on blood agar. In samples from diseased periodontal pockets, the recoveries of A. actinomycetemcomitans on A medium and H. aphrophilus on H medium equaled those on TSBV and TSBVF, respectively. In about 50% of the cultures on the reference media, contaminating bacteria were detected at levels higher than 10(4) CFU/ml of sample. The corresponding value for both A and H media was about 2%.
Simultaneous transmission of Borrelia burgdorferi and Babesia microti by individual nymphal Ixodes dammini ticks
Authors: Piesman J , Hicks T C , Sinsky R J , Obiri G
Published: January 2020

Nymphal Ixodes dammini ticks, selected from a group of ticks in which 22 of 31 (71%) contained dual Borrelia burgdorferi and Babesia microti infections, simultaneously transmitted B. burgdorferi and B. microti to 4 of 7 (57%) hamsters exposed to individual ticks.