bluuebunny/crossref_metadata_2025 huggingface dataset:
Isolation of a bovine rotavirus with a "super-short" RNA electrophoretic pattern from a calf with diarrhea
Authors: Paul P S , Lyoo Y S , Woode G N , Zheng S L , Greenberg H B , Matsui S , Schwartz K J , Hill H T
Published: January 2020

A rotavirus with a "super-short" RNA electropherotype was isolated from a calf with diarrhea and was designated VMRI strain. Segments 10 and 11 of this rotavirus migrated more slowly than did those of bovine rotavirus strains NCDV, B641, and B223. The electrophoretic pattern of the VMRI strain was similar to that reported for rotaviruses with super-short RNA electropherotypes from humans and rabbits. Northern (RNA) blot hybridization indicated that gene 11 of the VMRI strain was altered and migrated between gene segments 9 and 10. The subgroup of the VMRI strain was shown to be subgroup I. The VMRI strain of bovine rotavirus was neutralized by antisera containing polyclonal antibodies to rotavirus serotype 6 (bovine rotavirus serotype I) strains NCDV and B641 and by ascitic fluid containing monoclonal antibodies directed to VP7 of serotype 6 rotavirus. The VMRI strain was not neutralized by either polyclonal or monoclonal antibodies to strain B223 (bovine rotavirus serotype II). Collective data on the neutralization of the VMRI strain with monoclonal antibodies and polyclonal antibodies suggest that this virus is a member of the NCDV group (serotype 6) of rotaviruses (bovine rotavirus serotype I).
Identification of enterotoxigenic Escherichia coli isolates with enzyme-labeled synthetic oligonucleotide probes
Authors: Medon P P , Lanser J A , Monckton P R , Li P , Symons R H
Published: January 2020

Commercially available kits containing alkaline phosphatase-labeled oligonucleotide probes for Escherichia coli heat-stable enterotoxins (STI-H, STI-P, and STII) and the heat-labile enterotoxin were compared with bioassays and radiolabeled recombinant DNA probes to identify enterotoxigenic E. coli from 100 clinical isolates. There was very good agreement between the three methods.
The reason for loss of agglutinability of Pseudomonas aeruginosa cells heated to 60 degrees C
Authors: Liu P V
Published: January 2020

The loss of agglutinability of live Pseudomonas aeruginosa cells by mild heating to 60 to 80 degrees C is due to denaturation of surface slime, which can no longer combine with antibodies but is still attached to the cell surface and thus prevents access of antibodies to the cell wall. Prolonged boiling or autoclaving would not only denature the slime but also detach it from cell surface and thus make the cells accessible to antibodies directed to cell wall antigens. Heated cells are, however, no longer agglutinable by antibodies directed to slime antigens. After prolonged boiling or autoclaving, a large amount of polysaccharides appeared in the supernatant, and concomitantly, total cell volume as measured by turbidity of the cell suspension was significantly reduced.
DNA hybridization assay for congenital cytomegalovirus infection
Authors: Buffone G J , Demmler G J , Schimbor C M , Yow M D
Published: January 2020

An in vitro DNA hybridization assay was used to test 281 newborns for congenital infection with cytomegalovirus. The assay utilized an abbreviated method for DNA preparation and a dot blot assay that provided good sensitivity (100%) and specificity (98.9%) when compared with standard tissue culture, yet substantially reduced the total time of analysis. This assay would be a useful adjunct to tissue culture to diagnose newborns with congenital infection with cytomegalovirus.
Antithrombin III in the diagnosis of infection in febrile neutropenic patients
Authors: Kibbler C C , Prentice H G , Noone P
Published: January 2020

Antithrombin III levels in plasma were measured in 29 febrile neutropenic patients (31 episodes). They were lowest on day 2 in 62.5% of febrile patients and then increased or were normal in 81.3%. Low levels were present on day 2 in only 28.6% of patients with gram-negative infections, as compared with 33% of patients with doubtful infections.
Repetitive DNA sequences as probes for Mycobacterium tuberculosis
Authors: Eisenach K D , Crawford J T , Bates J H
Published: January 2020

Three cloned segments of Mycobacterium tuberculosis DNA which are promising as clinical probes were identified. An MboI digest of DNA from a clinical isolate of M. tuberculosis was cloned into bacteriophage M13. To identify recombinants specific for the M. tuberculosis complex, plaque lifts were hybridized with M. bovis and M. kansasii DNA. Recombinants which selectively hybridized with M. bovis DNA were characterized by probing slot blots and restriction digests of DNA from various mycobacteria. Three recombinants that did not hybridize to a significant extent with DNA from nontuberculous mycobacteria were identified. These three probes are of special interest because they are each repeated multiple (10 to 16) times in the M. tuberculosis chromosome. These probes were also shown to be useful for fingerprinting strains for epidemiological studies.
Laboratory detection of high-level aminoglycoside-aminocyclitol resistance in Enterococcus spp
Authors: Spiegel C A
Published: January 2020

Methods for detection of high-level resistance to aminoglycoside-aminocyclitol antibiotics were evaluated using 104 blood isolates of enterococci (97 Enterococcus faecalis and 7 Enterococcus faecium). Kanamycin was used to predict resistance to amikacin. Discrepancies between methods were resolved by time-kill studies. Four methods (MicroScan, macrotube, microtiter, and disk diffusion) for detecting resistance to gentamicin and streptomycin were compared, using 51 consecutive strains. There were 13 gentamicin-resistant strains, all of which were detected by macrotube, microtiter, and disk diffusion. MicroScan detected 2 (15%) of the 13. Of the 18 streptomycin-resistant strains, 17 (93%) were detected by disk diffusion, 16 (89%) by microtiter, 9 (50%) by macrotube, and 6 (33%) by MicroScan. An additional 53 consecutive strains were examined only by disk diffusion and microtiter for resistance to gentamicin, streptomycin, and kanamycin. The entire population of 104 strains contained 35 gentamicin-, 22 streptomycin-, and 54 kanamycin-resistant enterococcal isolates. All 35 gentamicin-resistant strains were detected by both methods. Of the 22 streptomycin-resistant strains, 1 was detected only by microtiter, 2 only by disk diffusion, and 19 by both methods. Of the 54 kanamycin-resistant strains, 1 was detected only by microtiter, 2 only by disk diffusion, and 51 by both methods. One additional strain which was resistant only by disk diffusion was susceptible to amikacin plus penicillin by time-kill studies. Disk diffusion is a suitable method for detection of high-level aminoglycoside-aminocyclitol resistance in E. faecalis and is well suited for sporadic testing. Additional data are necessary to determine the suitability of these tests for E. faecium.
Effect of high-speed rolling on herpes simplex virus detection and replication
Authors: Mavromoustakis C T , Witiak D T , Hughes J H
Published: January 2020

We examined the effect of high-speed rolling on herpes simplex virus replication. Inoculated cultures were rolled at 2, 96, or 383 rpm, while stationary cultures served as controls. At 24 h, inoculated cultures rolled at 96 rpm had a 6.8-fold increase in foci when compared with stationary cultures (P less than 0.01) and a 2.8-fold increase over cultures rolled at 2 rpm (P less than 0.05). Cultures rolled at 2 rpm had a 2.4-fold increase in foci over stationary cultures (P less than 0.05). Viral yield results correlated with focus results. Significantly more virus was present in cultures rolled at 96 rpm (7.3-fold) than in stationary cultures. Cultures rolled at 2 rpm produced 2.9-fold more virus than stationary cultures (P less than 0.05). Of 37 cultures rolled at 96 rpm, 33 (89%) were cytopathic effect positive at 96 h, while 18 of 40 (45%) were positive at 2 rpm and only 2 of 37 (5%) were positive for stationary cultures (P less than 0.01). Cultures rolled at 96 rpm produced maximum viral yields 2 days sooner than stationary cultures. Rolling of inoculated cultures should be used in the clinical laboratory to aid in the rapid detection of herpes simplex virus.
Human immune response to botulinum pentavalent (ABCDE) toxoid determined by a neutralization test and by an enzyme-linked immunosorbent assay
Authors: Siegel L S
Published: January 2020

To determine the immune status of persons receiving botulinum pentavalent (ABCDE) toxoid and to evaluate the effectiveness of the vaccine, we surveyed immunized individuals for neutralizing antibodies to type A and to type B botulinum toxins. After the primary series of three immunizations administered at 0, 2, and 12 weeks, 21 of 23 persons tested (91%) had a titer for type A that was greater than or equal to 0.08 international units (IU)/ml, and 18 (78%) had a titer for type B of greater than or equal to 0.02 IU/ml. (One international unit is defined as the amount of antibody neutralizing 10,000 mouse 50% lethal doses of type A or B botulinum toxin). Just before the first annual booster, 10 of 21 (48%) and 14 of 21 (67%) people lacked a detectable titer for type A and for type B, respectively. After the first booster, all individuals tested had a demonstrable titer to both types A and B. Of 77 persons who had previously received from one to eight boosts of the toxoid, 74 (96%) had an A titer of greater than or equal to 0.25 IU/ml and would not require an additional booster, according to the recommendations of the Centers for disease Control. However, only 44 of 77 (57%) had a B titer of greater than or equal to 0.25 IU/ml. In each group by booster number, even the group having had eight boosts, at least one person would require reimmunization on the basis of B titer. There was a wide range of antibody levels among individuals at the same point in the immunization scheme. Results from an enzyme linked immunosorbent assay, with purified type A or type B neurotoxin as the capture antigen, were compared with neutralization test results on 186 serum samples for type A and 168 samples for type B. Statistically, the correlation coefficients for results from the two assays were high (r = 0.69, P < 0.0001, for type A and r = 0.77, P < 0.0001, for type B). However, due to the wide dispersion of values obtained, using enzyme-linked immunosorbent assay results to predict neutralizing antibody levels is unwarranted.
Relationship between in vitro susceptibility test results for chloramphenicol and production of chloramphenicol acetyltransferase by Haemophilus influenzae, Streptococcus pneumoniae, and Aerococcus species
Authors: Matthews H W , Baker C N , Thornsberry C
Published: January 2020

Haemophilus influenzae, Streptococcus pneumoniae, and Aerococcus species were tested for susceptibility to chloramphenicol by standard broth microdilution and disk-diffusion methods. MICs and zone diameter breakpoints were correlated with production of chloramphenicol acetyltransferase (CAT). A comparison of MICs and zone diameters indicated that the interpretative criteria for H. influenzae and S. pneumoniae should be an MIC of less than or equal to 4 micrograms/ml or a zone diameter greater than or equal to 25 mm for susceptible strains and an MIC of greater than or equal to 8 micrograms/ml or a zone diameter of less than or equal to 20 mm for resistant strains; for Aerococcus species, interpretative criteria should be an MIC of less than or equal to 8 micrograms/ml or a zone diameter of greater than or equal to 20 mm for susceptible strains and an MIC of greater than or equal to 32 micrograms/ml or a zone diameter of less than or equal to 12 mm for resistant strains. All but four strains of H. influenzae and one strain of S. pneumoniae that were resistant to chloramphenicol by these criteria produced CAT. For Aerococcus species, however, chloramphenicol-resistant strains were negative for CAT as determined by a commercially available disk test. When comparing susceptibility results with CAT production, thiamphenicol was a better indicator of the presence of the enzyme than chloramphenicol and may be useful in assaying resistance to chloramphenicol.
Comparison of Dacron-tipped applicator and cytobrush for detection of chlamydial infections
Authors: Weiland T L , Noller K L , Smith T F , Ory S J
Published: January 2020

Endocervical samples were obtained from 435 females for Chlamydia detection by using both a Dacron swab and a cytobrush. Positive results were obtained from 35 swabs and 34 cytobrush specimens. All specimens positive with the cytobrush were detected also with swab samples. The cytobrush and Dacron swab appear to be comparable for the detection of endocervical infections with Chlamydia trachomatis.
Role of the 92.5-kilodalton outer membrane protein of Campylobacter jejuni in serological reactions
Authors: Jin T , Penner J L
Published: January 2020

Outer membrane proteins from 12 strains of Campylobacter jejuni were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with rabbit antisera prepared against Formalin-treated cell suspensions of three of the strains. The 92.5-kilodalton protein was confirmed to be a minor outer membrane protein and exhibited serological heterogeneity. Serological specificities of the proteins, however, were found not to correlate with serotypic specificities of the strains, as demonstrated by slide agglutination tests with whole-cell suspensions. The results suggested that the 92.5-kilodalton proteins were not likely to be participating antigenic determinants in the immunological reactions that give rise to agglutination of live-cell suspensions in the slide test.
Monoclonal antibodies to Mokola virus for identification of rabies and rabies-related viruses
Authors: Bussereau F , Vincent J , Coudrier D , Sureau P
Published: January 2020

Rabies and rabies-related virus strains were studied by using a panel of monoclonal antibodies directed against either nucleocapsid proteins or cell surface antigens of Mokola virus (Mok-3). Each strain was used in parallel to infect cultured cells and mice. Then, the patterns of reactivity of the different monoclonal antibodies were determined by the immunofluorescent-antibody staining procedure. On cells, the monoclonal antibodies differentiated fixed rabies virus strains (serotype 1) from rabies-related virus strains. The seven fixed strains (CVS, PV4, PM, Flury LEP and HEP, ERA, and SAD) reacted identically. The previous serotype groupings (serotype 2, Lagos-bat virus; serotype 3, Mokola virus; serotype 4, Duvenhage virus) established with anti-rabies monoclonal antibodies were confirmed, except for that of Lagos-bat Kindia, which appeared to be related to the African subtype of the Duvenhage serotype (Duv-2). Within the Mokola (Mok-1, -2, -3, and -5 and Umhlanga) and the Lagos-bat (Lag-1 and -2, Zimbabwe, Pinetown, and Dakar) serotypes, each strain appeared to be distinct. The African subtype of the Duvenhage serotype reacted differently from the European subtype. Within the Duvenhage serotype, subtypes Duv-4, -5, and -6 and Denmark reacted identically, while subtypes Duv-1, -2, and -3 and German Democratic Republic appeared to be distinct. The monoclonal antibodies specific for the cell surface antigens were also used in neutralization tests with all the strains. Two of them neutralized the infectivity of Mokola virus.
Enzyme-linked immunosorbent fluorescence assay and high-pressure liquid chromatography for analysis of humoral immune responses to Coxiella burnetti proteins
Authors: Schmeer N , Müller H P , Baumgärtner W , Wieda J , Krauss H
Published: January 2020

A microtiter enzyme-linked immunosorbent fluorescence assay based on alkaline phosphatase conjugate and 4-methylumbelliferyl phosphate as fluorogenic substrate was developed and adapted to quantitatively analyze immunoglobulin G subclass 1 (IgG1) and IgG2 responses of vaccinated and infected cattle to proteins of Coxiella burnetii. The enzyme-linked immunosorbent fluorescence assay surpassed the conventional enzyme-linked immunosorbent assay with a 50-fold-higher sensitivity and a broader range of linear dose-response signals. Antigens of C. burnetii were purified by sodium dodecyl sulfate extraction and molecular-sieve high-pressure liquid chromatography. The purified 14-, 27-, and 30-kilodalton proteins were used as antigens without any further treatment. Vaccination with either chloroform-methanol-extracted cell residues of C. burnetii or the 27-kilodalton major surface protein evoked an early IgG2 response to the 27-kilodalton protein (2 weeks after immunization), whereas IgG2 to lipopolysaccharides of C. burnetii was detected only in the late phase (13 weeks after immunization). These results may have implications for the serodiagnosis of acute and chronic Q fever. IgG1 against these antigens was induced solely by naturally occurring C. burnetii infections, indicating that infected cattle can be distinguished from vaccinated cattle by using the enzyme-linked immunosorbent fluorescence assay and SP27 antigen.
Diagnosis of toxoplasmic encephalitis in patients with acquired immunodeficiency syndrome by using a new serologic method
Authors: Suzuki Y , Israelski D M , Dannemann B R , Stepick-Biek P , Thulliez P , Remington J S
Published: January 2020

The present study was performed to develop a serological method for diagnosing toxoplasmic encephalitis in patients with acquired immunodeficiency syndrome (AIDS). The trophozoite form of Toxoplasma gondii, fixed with either Formalin or acetone, was used in a modification of an agglutination method previously shown to differentiate between the acute and the chronic (latent) stages of infection with toxoplasma in immunologically normal persons. By using these antigens in separate tests and evaluating the data for statistical significance, 70% of patients with AIDS with biopsy-proven toxoplasmic encephalitis were distinguished from control, ambulatory patients with AIDS with toxoplasma antibodies but without signs or symptoms of central nervous system involvement. In a separate study, the agglutination tests identified from controls 84% of patients with AIDS with two or more brain lesions detected by computed-tomographic or magnetic-resonance-imaging scans and suspected of having toxoplasmic encephalitis. Thus, these agglutination tests should prove valuable for the noninvasive diagnosis of toxoplasmic encephalitis in patients with AIDS.
Comparison between radioimmunoassay and direct and indirect enzyme-linked immunosorbent assays for determination of antibodies against Haemophilus influenzae type b capsular polysaccharide
Authors: Lagergård T , Trollfors B , Claesson B A , Schneerson R , Robbins J B
Published: January 2020

Levels of antibodies against Haemophilus influenzae type b capsular polysaccharide were determined in acute-phase and convalescent-phase serum samples obtained from 21 children with invasive H. influenzae type b infections and from 44 children vaccinated with two H. influenzae type b vaccines. Amounts of immunoglobulin G (IgG), IgM, and IgA antibodies were measured by direct and indirect enzyme-linked immunosorbent assay (ELISA), and the total amount of antibodies was measured by radioimmunoassay (RIA). Results obtained by ELISA were calculated by multiple-point parallel-line comparison and by endpoint analysis. A very good correlation was obtained between direct and indirect ELISA values. In the lower range of antibody concentrations, the correlation between ELISA values obtained by endpoint analysis and those obtained by multiple-point parallel-line comparison was poor, since the latter method of calculation yielded values of up to 1 microgram/ml in sera that were negative according to endpoint analysis. These sera with negative endpoint titers also had undetectable or very low antibody concentrations as measured by RIA. Consistent with this finding, in acute-phase and prevaccination sera with undetectable or low antibody concentrations as measured by RIA, ELISA values calculated by multiple-point parallel-line comparison were much higher. In sera with higher antibody concentrations, however, parallel-line comparisons showed good correlation between RIA and ELISA values. Although no reference method for measuring true antibody concentrations is available, ELISA values as calculated by multiple-point parallel-line comparison appear to overestimate antibody concentrations in sera containing low antibody concentrations, whereas ELISA values obtained by endpoint analysis are less well correlated with RIA values at higher concentrations.
Use of supplemented Stainer-Scholte broth for the isolation of Bordetella pertussis from clinical material
Authors: von Koenig C H , Tacken A , Finger H
Published: January 2020

The use of Stainer-Scholte broth supplemented with (2,6-O-dimethyl)beta-cyclodextrin (heptakis) for the isolation of Bordetella pertussis from clinical specimens was evaluated with 3,632 nasal swabs from children and adults with suspected whooping cough or from their family contacts. The liquid enrichment medium was subcultured on charcoal agar with 10% defibrinated horse blood. Charcoal agar and soft charcoal agar served as the standard procedure to detect B. pertussis. We isolated 772 strains of B. pertussis (21%). Charcoal agar alone detected 87% of all strains (n = 668), soft charcoal agar grew 78% (n = 602), and 637 strains (83%) were isolated when Stainer-Scholte broth with heptakis was used. We detected 590 isolates with all three media. Whereas 65 strains grew only on charcoal agar, 27 strains were detected by soft charcoal agar. Supplemented Stainer-Scholte broth allowed the isolation of an additional 77 strains which did not primarily grow on charcoal media (P less than 0.05). Our data indicate that Stainer-Scholte medium supplemented with heptakis can be effectively used as an enrichment medium for detection of B. pertussis in clinical specimens.
Calculation of antibody affinity in homogeneous and heterogeneous systems
Authors: Chalquest R R
Published: January 2020

Antibody affinity is an important determinant of all antibody-antigen reactions. A new computer program, AFCRV, was developed to calculate binding constants with data from a radioimmunoassay on most microcomputers in the laboratory by using constant-ratio dilution curves. Evaluation of a homogeneous or heterogeneous antibody in the presence of a single antigen can be accomplished.
Phosphatase activity is a constant feature of all isolates of all major species of the family Enterobacteriaceae
Authors: Satta G , Pompei R , Grazi G , Cornaglia G
Published: January 2020

In this study we evaluated phosphatase activity in members of the family Enterobacteriaceae by conventional methods and by a novel method. The novel method is based on the formation of bright-green-strained colonies by phosphatase-positive, but not phosphatase-negative, strains in the presence of a phosphate substrate, such as phenolphthalein monophosphate or 6-benzoylnaphthyl phosphate (6-BNP), and methyl green. A total of 1,055 strains belonging to 65 different species of Enterobacteriaceae were tested for green staining of the colonies in the presence of methyl green and either phenolphthalein monophosphate or 6-BNP and for phosphatase activity by three different conventional methods. With the sole exception of one Leminorella richardii type strain, all isolates of all of the species formed green-stained colonies in the presence of the substrate 6-BNP. All strains were phosphatase positive by all of the conventional methods.
Campylobacter hyointestinalis: an opportunistic enteropathogen?
Authors: Minet J , Grosbois B , Megraud F
Published: January 2020

A new case of Campylobacter hyointestinalis-associated diarrhea in a human is reported. The medical history of the patient was significant for immunodeficiency because of an evolutive chronic myeloid leukemia. The diarrhea rapidly stopped after administration of oral erythromycin. No other enteropathogenic agent was found by routine examination of stools. Although neither serology nor autopsy was available, this observation appears to be suggestive of the possible enteropathogenicity of C. hyointestinalis for patients at risk.
Identifying Mycobacterium tuberculosis cultures by gas-liquid chromatography and a computer-aided pattern recognition model
Authors: Maliwan N , Reid R W , Pliska S R , Bird T J , Zvetina J R
Published: January 2020

A procedure that uses gas-liquid chromatography and a pattern recognition computer model was developed for distinguishing cultures of Mycobacterium tuberculosis from cultures of other mycobacteria, common bacteria, and fungi. In this procedure, a sample of a culture preparation is methanolyzed and trimethylsilylated sequentially and injected into a gas chromatograph equipped with a flame ionization detector. A pattern recognition procedure computes an error score by comparing the gas-liquid chromatography peak responses of a culture to those of a standard M. tuberculosis culture. Ten M. tuberculosis cultures were used in the development of the pattern recognition model. Computed error scores of 5 or less were established for identifying an M. tuberculosis culture. The method was evaluated with two sets of test samples, non-M. tuberculosis and M. tuberculosis cultures. Sample identification was correct for all 14 M. tuberculosis cultures (M. tuberculosis or non-M. tuberculosis), 45 fungal cultures, 94 bacterial cultures, and all but 1 of 18 cultures of mycobacteria other than tuberculosis (MOTT). The false prediction represented an isolate of M. fortuitum. For M. tuberculosis, fungal, bacterial, and MOTT cultures, the ranges of error scores were 1 to 5, 16 to 33, 13 to 34, and 4 to 26, respectively. Therefore, we have demonstrated that this diagnostic model can distinguish M. tuberculosis from non-M. tuberculosis cultures with a high degree of accuracy.
Comparative study of inactivation of herpes simplex virus types 1 and 2 by commonly used antiseptic agents
Authors: Croughan W S , Behbehani A M
Published: January 2020

A comparative study of the different reactions of herpes simplex virus types 1 and 2 to Lysol, Listerine, bleach, rubbing alcohol, Alcide disinfectant (Alcide Corp., Westport, Conn.), and various pHs, temperatures, and UV light exposures was performed. Both types of stock virus (titers of approximately 10(6) and 10(5.5) for types 1 and 2, respectively) were inactivated by 0.5% Lysol in 5 min; by Listerine (1:1 mixtures) in 5 min; by 2,000 ppm (2,000 microliters/liter) of bleach in 10 min; by rubbing alcohol (1:1 mixtures) at zero time; by Alcide disinfectant (0.2 ml of virus plus 2.0 ml of Alcide) at zero time; by pHs 3, 5, and 11 in 10 min; and by a temperature of 56 degrees C in 30 min. A germicidal lamp (model G30TB; General Electric Co., Schenectady, N.Y.) (30 W) at a distance of 48 cm failed to completely inactivate the two types in 15 min. Type 1 showed slightly more resistance to Listerine and bleach and significantly more resistance to heat; moreover, pH 9 did not affect the infectivity of either type after 10 min.
Genetic variants of influenza A/Taiwan/1/86 cocirculating in Canada during the winter of 1986 to 1987
Authors: Brown E G
Published: January 2020

The first isolate of influenza virus in Canada during the winter of 1986 to 1987 was a genetic variant of A/Taiwan/1/86. This genetic variant type was the predominant strain obtained from several of the western provinces. The variant strains were antigenically indistinguishable from A/Taiwan/1/86 but were remarkably distinct by T1 oligonucleotide mapping. T1 mapping of individual genome segments indicated that the variants evolved from an A/Taiwan/1/86-like virus through the accumulation of point mutation or deletion or insertion events and probably do not contain foreign genes. The relative distribution of genetic variation was approximately equal among the individual genes, with the possible exception of segments 1 or 2 that were analyzed in combination and thus could not be individually associated with the observed variation.
Serotypes of Ureaplasma urealyticum isolated from normal pregnant women and patients with pregnancy complications
Authors: Naessens A , Foulon W , Breynaert J , Lauwers S
Published: January 2020

To compare the distribution of Ureaplasma urealyticum serotypes 1 to 10 in different patient populations, the serotypes of 240 U. urealyticum strains from 207 patients were determined by the indirect immunofluorescence test by using U. urealyticum antisera 1 to 10. Strains were obtained from the following four patient groups: group 1, 24 couples in which the women had a history of recurrent spontaneous abortion; group 2, 25 patients who had their first spontaneous abortion; group 3, 14 pregnant patients with pregnancy complications (premature delivery, intrauterine death); and group 4, 138 patients with uneventful pregnancies. The serotypes most often found in these 207 patients were as follows: serotype 3, 52.2%; serotype 6, 30.3%; serotype 10, 11.4%; serotype 1, 9.5%; serotype 4, 6.5%; serotype 8, 6.5%. Serotypes 2, 5, 7, and 9 were found in less than 1% of the patients. More than one serotype was found in 16.9% of the patients. The overall distribution of the 10 serotypes in the different groups was similar, except for that of serotype 4. Serotype 4 was isolated from 20.8% of the patients in group 1 and from 5.1% of the patients in group 4 (P less than 0.01). Results of this study indicate that U. urealyticum serotype 4 can be isolated more frequently from patients with a history of recurrent miscarriages than from normal pregnant women.
Comparable sensitivities for detection of human immunodeficiency virus by sensitive reverse transcriptase and antigen capture enzyme-linked immunosorbent assays
Authors: Lee M H , Sano K , Morales F E , Imagawa D T
Published: January 2020

The sensitive overnight reverse transcriptase and the antigen capture enzyme-linked immunosorbent assays were tested for their ability to detect human immunodeficiency virus. The two assays were able to quantitate as few as 75 to 130 virus particles. Thus, both tests offer similar sensitivities for determining the presence of the virus.
Characterization of cross-reactive proteins detected by Culturette Brand Rapid Latex Test for Clostridium difficile
Authors: Lyerly D M , Ball D W , Toth J , Wilkins T D
Published: January 2020

Clostridium sporogenes, Peptostreptococcus anaerobius, and Bacteroides asaccharolyticus have been reported to react in the Culturette Brand Rapid Latex Test (Marion Scientific, Div. Marion Laboratories, Inc., Kansas City, Mo.) for Clostridium difficile. From the results of this study we showed that C. sporogenes and P. anaerobius produce a protein which is very similar biochemically and immunologically to the protein of C. difficile that is detected by the test. Thus, the positive latex reactions observed with C. sporogenes and P. anaerobius are due to a cross-reactive protein. We did not detect this cross-reactive protein in filtrates from B. asaccharolyticus, indicating that this bacterium reacts with the latex reagent by some other mechanism. We cloned the C. difficile gene that codes for the cross-reactive protein and showed that the protein produced by the recombinant organism is nontoxic and distinct from toxin A, thus confirming our earlier findings.
Evaluation of a rapid tube assay for presumptive identification of Escherichia coli from veterinary specimens
Authors: Iritani B , Inzana T J
Published: January 2020

Three hundred sixty-six isolates of gram-negative, oxidase-negative bacteria from veterinary specimens were tested by a tube test for identification as Escherichia coli by production within 60 min of indole, beta-galactosidase, and beta-glucuronidase. The test correctly identified 255 of 269 isolates of E. coli (95% sensitivity) and correctly indicated that 97 of 97 isolates were not E. coli (100% specificity). We conclude that production of indole, beta-galactosidase, and beta-glucuronidase as measured by a rapid tube test is useful for identification of E. coli from veterinary specimens.
Comparison of Track XI fluorometric immunoassay with Bio-EnzaBead enzyme-linked immunosorbent assay for detection of serum antibody to mouse hepatitis virus
Authors: La Regina M C , Lonigro J , Woods L , Hall W C , Doyle R E
Published: January 2020

The Track XI system (Microbiological Associates, Bethesda, Md.) was compared with the Bio-EnzaBead assay (Organon Teknika, Durham, N.C.) for the detection of antibody to mouse hepatitis virus (MHV). Strain A/J mice were inoculated intranasally with MHV type 3. Sera were collected at 1, 2, 4, and 9 weeks postinoculation and tested. Individual serum samples were retested twice by each method. The results suggested that the Track XI system was more sensitive and reliable than the Bio-EnzaBead assay in detecting antibody to MHV type 3 in individual serum samples from A/J mice.
Yersinia pseudotuberculosis infection contracted through water contaminated by a wild animal
Authors: Fukushima H , Gomyoda M , Shiozawa K , Kaneko S , Tsubokura M
Published: January 2020

We performed epidemiological studies on Yersinia pseudotuberculosis in one valley where a 3-year-old boy had been infected with Y. pseudotuberculosis serotype 4b in December 1982. Y. pseudotuberculosis serotype 4b was isolated from a water sample derived from a mountain stream from which the boy had drunk and from 1 of 41 rats trapped in the upper part of this stream in December 1986. The restriction endonuclease patterns of the plasmids in these isolates showed the rat and patient isolates to be identical but distinct from the water isolate. These data suggest the potential for transmission of Y. pseudotuberculosis through water contaminated by nondomesticated animals carrying this species.
Detection of respiratory syncytial virus in clinical specimens by viral culture, direct and indirect immunofluorescence, and enzyme immunoassay
Authors: Hughes J H , Mann D R , Hamparian V V
Published: January 2020

We evaluated prospectively the detection of respiratory syncytial virus (RSV) by culture and by direct antigen detection using an indirect immunofluorescence assay (IFA), a direct monoclonal immunofluorescence assay (DFA), and a monoclonal enzyme immunoassay (EIA). Of 221 specimens, 95 (43%) were culture positive for RSV, 4 (1.8%) contained more than one virus, and 17 (7.6%) contained a virus other than RSV. Overall, HEp-2 and Flow 6000 cells grew significantly more RSV isolates (82 and 72%, respectively) than A549 cells, which grew only 29% of the isolates. The mean time for RSV detection with HEp-2 cells was 2.9 days. This was significantly less than the mean time for RSV detection with either Flow 6000 cells (6.1 days) or A549 cells (6.4 days). Of 221 specimens, 129 were tested simultaneously by culture, IFA, and DFA. Of these 129 specimens, 62 (48%) were positive by culture, 69 (53%) were positive by IFA, and 70 (54%) were positive by DFA. For 92 specimens screened simultaneously by culture, IFA, and EIA, positive results were obtained for 33 (36%) of the specimens by both culture and IFA and for 29 (32%) of the specimens by EIA. Of 126 culture-negative specimens, 21 (17%) were positive for RSV when determined by IFA. Conversely, 14 (15%) of 95 RSV culture-positive specimens were negative by IFA, whereas DFA missed 19% of the culture-positive specimens. Compared with culture, the Kallestad EIA kit had a sensitivity and specificity of 73 and 92% respectively, but missed 9 (27%) of 33 culture-positive specimens. These data demonstrate that isolation by culture continues to be important for viral diagnosis of REV infections and that for valid comparative studies between viral isolations and rapid detection methods, both sensitive host cells and appropriate test conditions are required.
Achromobacter xylosoxidans (Alcaligenes xylosoxidans subsp. xylosoxidans) bacteremia associated with a well-water source: case report and review of the literature
Authors: Spear J B , Fuhrer J , Kirby B D
Published: January 2020

A case of community-acquired Achromobacter xylosoxidans bacteremia in a patient with metastatic breast carcinoma is described. The patient's home drinking water was identified as the source of her bacteremia. The case represents the first in which a community-acquired infection due to this organism has been attributed to a documented water source.
Positive reaction in mouse ligated intestinal loop assay with nonenterotoxigenic and nonhemolytic strains of Staphylococcus aureus
Authors: Sakaguchi T , Sakaguchi S , Nakamura I , Kudo Y , Ichiman Y , Yoshida K
Published: January 2020

Nonenterotoxigenic and nonhemolytic strains of Staphylococcus aureus exhibited a positive reaction in the mouse intestinal loop assay except for a strain negative for the egg yolk reaction. Edema and swelling in all positive loops, increased bacterial cell numbers within intestinal loops, and extremely close adhesion to or inclusion of numerous bacterial cells in HeLa cells after inoculation of the strains were observed. These results suggest a possible invasive ability of the strains.
Characterization of clinically significant isolates of Staphylococcus epidermidis from patients with endocarditis
Authors: Etienne J , Brun Y , el Solh N , Delorme V , Mouren C , Bes M , Fleurette J
Published: January 2020

Biotyping, slime production, bacteriophage typing, serotyping, antibiograms, and plasmid profiles were used to characterize 19 Staphylococcus epidermidis strains isolated from 12 patients with prosthetic valve endocarditis and from 7 patients with native valve endocarditis. With the API Staph battery, 12 different biocodes with, at the most, three differences were obtained. Slime production was found for 10 strains (53%). Agglutinogens investigated by agglutination with two specific sera were found for 12 strains (63.1%). Three strains were phage typable (15.2%). Against a panel of nine antimicrobial agents, 15 different profiles were found. Multiply antibiotic-resistant strains were isolated from patients with prosthetic valve endocarditis when disease onset occurred less than 18 months after heart surgery and from patients with native valve endocarditis who received antibiotics immediately prior to their illness. All of the strains were available for plasmid analysis, and all the DNA profiles were distinct. On gels run in Tris-borate buffer, 73.7% of the strains had large plasmids of more than 30 megadaltons. A small plasmid of 2.8 megadaltons was found in multiply resistant strains and in strains resistant only to tetracyclines. None of the isolates appeared to be the same strain, and the bacteriological differences between the strains were confirmed mainly by the antibiotic susceptibility profile and the plasmid pattern analysis. These bacteriological results were in agreement with the clinical data.
Characterization of Rhodococcus equi-like bacterium isolated from a wound infection in a noncompromised host
Authors: Müller F , Schaal K P , von Graevenitz A , von Moos L , Woolcock J B , Wüst J , Yassin A F
Published: January 2020

A case of superficial wound infection in a noncompromised host due to a Rhodococcus equi-like organism, acquired from soil, is described. The strain grew poorly at 37 degrees C and showed several differences from wild-type R. equi in biochemical tests and in its fatty acid and mycolic acid compositions.
Microbiology of indwelling central intravascular catheters
Authors: Haslett T M , Isenberg H D , Hilton E , Tucci V , Kay B G , Vellozzi E M
Published: January 2020

Indwelling central-line catheters (n = 502) from 362 patients in intensive care units were analyzed prospectively. The skin site, 6-cm-distal and -proximal subcutaneous segments of the catheter, exudates, and blood were cultured. Semiqualitative roll plate cultures of the catheter segments were followed by broth cultures and examined for 72 h. All isolates were identified, and susceptibilities were determined. Line infections, defined clinically, yielded 22 different microbial species; 10 different species were recovered from colonized lines. Of the Staphylococcus epidermidis isolates recovered, 39% occurred singly and 21% occurred in combination with other microorganisms; Enterococcus faecalis, S. hominis, and Pseudomonas aeruginosa were next in frequency of isolation. Line infections also yielded other staphylococci, viridans group streptococci, several members of the family Enterobacteriaceae, acinetobacters, anaerobic bacteria, Candida spp., and Aspergillus fumigatus. While S. epidermidis was also the most frequent isolate among the line colonizers, different species and different frequencies of isolation were found among this group. The study showed that the distal catheter segment broth culture was the best predictor of clinical line infections; in addition, gram-negative bacteria were isolated only from the catheters of patients with overt infections.
Xylohypha emmonsii sp. nov., a new agent of phaeohyphomycosis
Authors: Padhye A A , McGinnis M R , Ajello L , Chandler F W
Published: January 2020

Xylohypha emmonsii Padhye, McGinnis et Ajello is described as a new dematiaceous opportunistic pathogen of humans and lower animals. This hyphomycete differs from X. bantiana (Saccardo) McGinnis, Padhye, Borelli et Ajello by its inability to grow at 40 degrees C and by the development of short chains of asymmetrically bent to ellipsoid blastoconidia. In brain tissue of mice, experimentally infected with X. emmonsii, the host response to the invading fungus was similar to that observed in mice infected with X. bantiana. In tissues X. bantiana occurred predominantly as long, septate hyphae, in contrast to X. emmonsii, which was more polymorphic.
Comparison of serum and mucosal antibody responses following severe acute rotavirus gastroenteritis in young children
Authors: Grimwood K , Lund J C , Coulson B S , Hudson I L , Bishop R F , Barnes G L
Published: January 2020

The development of mucosal immunity is presumed to be the most important marker of rotavirus infection. The practical difficulties of obtaining small-bowel secretions stimulated this study of the antibody response to acute rotavirus infection at other sites. Forty-four infants admitted to the hospital with rotavirus gastroenteritis had serum, saliva, and feces collected at the acute phase (median, 5.5 days), during convalescence (median, 33.5 days), and 4 months later (median, 12.2 weeks). A subgroup of 19 children also had duodenal juice collected in parallel. Rotavirus-specific immunoglobulin G (IgG), IgA, secretory immunoglobulin, and IgM were measured and compared in all samples. The results showed that the estimation of antirotavirus serum IgM, serum IgG, duodenal juice IgA, and duodenal juice IgM by an enzyme immunoassay indicated an immune response to severe primary rotavirus infection in all children. Four months later, the levels of serum IgG and IgA served as the most sensitive markers of the preceding rotavirus infection. The predictive accuracies of immune responses at different sites in relation to a positive IgA immune response in the duodenum were calculated. Fecal IgA predicted duodenal IgA rotavirus antibodies with accuracies of 86% at 1 month and 92% at 4 months. The high sensitivity of serum IgM and IgG in detecting rotavirus infection and the high predictive accuracy of fecal IgA as an indicator of duodenal IgA abrogates the need for duodenal intubation to detect (or monitor) an immune response to rotavirus infection. This finding has important practical implications for epidemiological studies of acute diarrhea in children and in rotavirus vaccine trials.
Role of specific immunoglobulin E to excretory-secretory antigen in diagnosis and prognosis of hookworm infection
Authors: Ganguly N K , Mahajan R C , Sehgal R , Shetty P , Dilawari J B
Published: January 2020

Total immunoglobulin E (IgE) and specific IgE were assayed by a radioimmunosorbent test and a reverse enzyme immunoassay in patients with hookworm infections before and after treatment. A total of 77 subjects (30 patients with hookworm infections and 47 subjects as controls) were studied. Both specific and total IgE levels in serum and jejunal juice were raised in hookworm patients. There was a significant decrease in IgE levels after therapy. Total IgE levels were raised in other nematode infections, but specific IgE levels were low. The reverse enzyme immunoassay for specific IgE was highly specific (96%) and sensitive (100%) and may be used in the serodiagnosis of hookworm infections.
Prevention of pneumococcal otitis media in chinchillas with human bacterial polysaccharide immune globulin
Authors: Shurin P A , Giebink G S , Wegman D L , Ambrosino D , Rholl J , Overman M , Bauer T , Siber G R
Published: January 2020

Clinical and experimental observations suggest that immune globulin may prevent otitis media (OM) in children. We performed experiments in chinchillas to test the hypothesis that human bacterial polysaccharide immune globulin (BPIG) might prevent OM caused by Streptococcus pneumoniae. Animals were given BPIG or saline intraperitoneally on day 0. On day 3 the epitympanic bulla was inoculated with S. pneumoniae type 7F. All 12 saline-treated and none of 12 BPIG-treated animals developed pneumococcal OM by day 7 (P less than 0.0001). Bacteremia developed in 6 of 12 saline- and 0 of 12 BPIG-treated animals (P = 0.007). Death with pneumococcal OM occurred within 28 days in 5 of 12 saline- and 0 of 12 BPIG-injected animals (P = 0.02). A chinchilla-specific immunoassay was used to show that surviving saline-injected animals developed serum anticapsular antibody; BPIG-treated animals had no detectable response. At levels of anticapsular immunoglobulin G similar to those of human adults, BPIG given systemically prevented pneumococcal OM and disseminated infection in chinchillas. BPIG may be of value in preventing human bacterial infection and may also inhibit development of antibody if it affects local infection or colonization. Specific immunoglobulin G antibody may provide an important antibacterial defense of mucosal surfaces of the respiratory tract.
Evaluation of a nonisotopically labeled oligonucleotide probe to detect the heat-stable enterotoxin gene of Escherichia coli by the DNA colony hybridization test
Authors: Nishibuchi M , Arita M , Honda T , Miwatani T
Published: January 2020

A commercially available, alkaline-phosphatase-conjugated oligonucleotide probe for detecting the heat-stable enterotoxin gene of Escherichia coli was compared with cloned gene probes by examining E. coli isolates from traveler's diarrhea by DNA colony hybridization tests. The oligonucleotide probe was useful in specifically identifying the so-called STh gene. No deproteinization of sample was necessary to prepare the colony blots.
New plate medium for screening and presumptive identification of gram-negative urinary tract pathogens
Authors: Thaller M C , Berlutti F , Dainelli B , Pezzi R
Published: January 2020

A new selective, differential plating medium to screen the common gram-negative urinary tract pathogens is described. The medium combines adonitol fermentation, phenylalanine deaminase, and beta-glucuronidase tests and allows the indole and cytochrome oxidase tests to be performed directly from the plates. High-level agreement with individual conventional tests was recorded in comparative studies with 504 cultures of gram-negative rods. There was 100% agreement, except for the Providencia spp. indole spot test (61.6% agreement). Adonitol fermentation by Providencia species could not be determined. Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, and Pseudomonas aeruginosa were identified with a high efficiency (100, 85.7, 83.5, and 100% agreement, respectively) without further testing. There was 96% overall agreement for the 267 infected urine samples tested.
Purification and properties of Bacteroides heparinolyticus heparinase (heparin lyase, EC 4.2.2.7)
Authors: Nakamura T , Shibata Y , Fujimura S
Published: January 2020

Heparinase (heparin lyase, EC 4.2.2.7) was isolated from the cell extract of an oral bacterium, Bacteroides heparinolyticus. It was a basic protein with an isoelectric point of 9.5. Its molecular weight was 63,000. The enzyme was the most active against heparin among the tested mucopolysaccharides. Catalytic properties may be similar to those of heparinase of Flavobacterium heparinum, since the enzymatic degradation products obtained by using the two enzymes were the same on the basis of paper chromatography.