bluuebunny/crossref_metadata_2025 huggingface dataset:
Detection of an immunoglobulin M response in the elderly for early diagnosis of respiratory syncytial virus infection
Authors: Vikerfors T , Grandien M , Johansson M , Pettersson C A
Published: January 2020

The indirect fluorescent-antibody technique was compared with indirect and mu-capture enzyme-linked immunosorbent assays for the detection of respiratory syncytial virus (RSV) immunoglobulin M (IgM) in the elderly. Sera from 47 patients (mean age, 70 years) with acute lower respiratory tract infections caused by RSV were investigated. Specific IgM was detected in 81% (38 of 47) of the patients. The fluorescent-antibody technique, which gave 70% positive results, proved to be the most sensitive of the three methods. An IgM response was seldom seen in sera from the elderly within the first week of disease, but was present in 85% of sera (33 of 39) collected between days 11 and 30 of disease. In some patients it persisted for more than 6 weeks. Detection of IgM was found to be a useful tool for the diagnosis of RSV infections in elderly patients.
Evaluation of MicroScan MIC panels for detection of oxacillin-resistant staphylococci
Authors: Woods G L , Yam P
Published: January 2020

Clinical isolates of staphylococci (420 Staphylococcus aureus isolates and 248 coagulase-negative staphylococci) were tested by both MicroScan MIC panels (MicroScan, West Sacramento, Calif.) and an oxacillin agar screen (Mueller-Hinton agar [Difco Laboratories, Detroit, Mich.] containing 6 micrograms of oxacillin per ml and 4% NaCl) to evaluate the ability of MicroScan to detect oxacillin-resistant strains. MicroScan panels and oxacillin agar screen plates were incubated at 35 degrees C for 24 h and at 30 degrees C for an additional 24 h. Endpoints were recorded at 24 and 48 h. By MicroScan, 23 (5.5%) and 30 (7%) S. aureus isolates and 161 (65%) and 162 (65%) coagulase-negative staphylococci were oxacillin resistant at 24 and 48 h, respectively. At both 24 and 48 h, 23 (5.5%) S. aureus isolates and 162 (65%) coagulase-negative staphylococci were resistant by the oxacillin agar screen. Five strains for which the oxacillin MIC was 2 or 4 micrograms/ml and eight strains resistant to oxacillin only at 48 h were further evaluated by broth macrodilution testing for oxacillin with and without clavulanic acid, by oxacillin and amoxicillin-clavulanic acid disk diffusion, and by oxacillin agar screen comparing Mueller-Hinton agars purchased from Difco and BBL Microbiology Systems, Cockeysville, Md. By this additional testing, all 10 S. aureus isolates and 1 of 3 coagulase-negative staphylococci examined produced increased amounts of beta-lactamase. One coagulase-negative staphylococcus appeared to be truly intermediately oxacillin susceptible. There was no significant difference in the rate of detection of oxacillin resistance between MicroScan and the agar screen. MicroScan panels should be incubated for 24 h only, because prolonged incubation caused strains producing excessive amounts of beta-lactamase to appear to be falsely oxacillin resistant.
Variation of electrophoretic karyotypes among clinical isolates of Candida albicans
Authors: Merz W G , Connelly C , Hieter P
Published: January 2020

Orthogonal-field-alternation gel electrophoresis was used to compare clinical isolates of Candida albicans by resolving chromosome-sized DNA molecules into an electrophoretic karyotype. Seven to nine bands were observed among isolates recovered from 17 patients. In addition, 14 distinct electrophoretic patterns were noted among the isolates from these patients. In a given individual, isolates were likely to have identical electrophoretic patterns. Therefore, the electrophoretic karyotype patterns demonstrated by orthogonal-field-alternation gel electrophoresis can be used to designate a strain for epidemiologic studies.
Monoclonal antibodies to Pasteurella haemolytica serotype 1 lipopolysaccharide: demonstration of antigenic similarities among several serotypes
Authors: Durham J A , Antone S M , Cunningham M W , Confer A W
Published: January 2020

Murine monoclonal antibodies (MAbs) were produced which were specific for Pasteurella haemolytica serotype 1 lipopolysaccharide (LPS). The MAbs also reacted with LPS present in a partially purified antigen derived from a saline extract of the organism. The epitope to which the MAbs were directed was a carbohydrate which was sensitive to oxidation with periodate, had a molecular weight between 14,000 and 25,000 as determined by immunoblotting, and was present in a crude O-antigen preparation of P. haemolytica LPS. The MAbs did not react with purified capsular polysaccharide from P. haemolytica serotype 1. In an enzyme-linked immunosorbent assay, reaction of the MAbs with LPS obtained from 14 gram-negative bacteria failed to detect any cross-reactivity with P. haemolytica LPS. However, the MAbs detected antigenic similarities among P. haemolytica serotypes 1, 5, 6, 7, 8, and 12 and, to a lesser extent, 4 and 14. These studies indicate that the LPS-O-antigens from several P. haemolytica serotypes have similar epitopes and may be partially responsible for shared antigenicity among serotypes.
The urinary bladder, a consistent source of Borrelia burgdorferi in experimentally infected white-footed mice (Peromyscus leucopus)
Authors: Schwan T G , Burgdorfer W , Schrumpf M E , Karstens R H
Published: January 2020

White-footed mice, Peromyscus leucopus, were experimentally infected in the laboratory with Borrelia burgdorferi, the causative agent of Lyme disease. After mice were infected by intraperitoneal or subcutaneous inoculation or by tick bite, attempts were made to culture spirochetes from the urinary bladder, spleen, kidney, blood, and urine. Spirochetes were most frequently isolated from the bladder (94%), followed by the kidney (75%), spleen (61%), and blood (13%). No spirochetes were isolated from the urine. Tissue sectioning and immunofluorescence staining of the urinary bladder demonstrated spirochetes within the bladder wall. The results demonstrate that cultivation of the urinary bladder is very effective at isolating B. burgdorferi from experimentally infected white-footed mice and that culturing this organ may be productive when surveying wild rodents for infection with this spirochete.
Prevalence and persistence of Neisseria cinerea and other Neisseria spp. in adults
Authors: Knapp J S , Hook E W
Published: January 2020

Neisseria cinerea is a commensal Neisseria sp. which was first described in 1906 but was subsequently misclassified as a subtype of Branhamella catarrhalis. N. cinerea resembles Neisseria gonorrhoeae in both cultural and biochemical characteristics and, thus, may also have been misidentified as N. gonorrhoeae. Of 202 patients whose oropharynges were colonized by Neisseria spp., N. cinerea was isolated in 57 (28.2%) patients, including 25 (30.1%) of 83 women, 22 (23.9%) of 92 heterosexual men, and 10 (37.0%) of 27 homosexual men in Seattle, Wash., in 1983 to 1984. N. cinerea was isolated from the urethra of only one (1.1%) patient. The oropharynges of many individuals were colonized persistently by strains of N. cinerea and other Neisseria spp.
Isolation and restriction endonuclease cleavage of Anaplasma marginale DNA in situ in agarose
Authors: Krueger C M , Buening G M
Published: January 2020

Bacterial restriction endonucleases were used to produce DNA cleavage patterns that could be useful as tools to study the relatedness among Anaplasma marginale isolates. Bovine erythrocytes infected with A. marginale were lysed, washed, and embedded in agarose. The embedded erythrocytes and bacterial pathogens were partially digested by sequential infiltration of the agarose with acetone, lysozyme, sodium dodecyl sulfate, and proteinase K. The unfragmented genomic DNA was left supported and protected in a porous matrix. The DNA was digested in situ in agarose under the following conditions: (i) brief treatment with phenol, (ii) brief washing with distilled water, and (iii) adjustment of restriction enzyme digestion mixture to compensate for the volume of the agarose. The cleaved DNA was electrophoresed horizontally to produce a DNA cleavage pattern. Of 19 restriction enzymes screened, 12 produced distinct DNA bands from the genomes of each of the five A. marginale isolates examined. The DNA cleavage pattern produced from each isolate with a given restriction enzyme was reproducible. However, the DNA cleavage patterns produced from different isolates with a given restriction enzyme were not necessarily identical. This procedure could be modified for general bacterial DNA isolation, in situ agarose digestion, and manipulations.
Characterization of enteroinvasive Escherichia coli strains isolated from children with diarrhea in Chile
Authors: Faundez G , Figueroa G , Troncoso M , Cabello F C
Published: January 2020

Analysis of stool samples from 912 cases of diarrhea among Chilean infants and 1,112 controls resulted in the isolation of 17 enteroinvasive Escherichia coli (EIEC) strains from diarrhea cases (1.9%) and 3 EIEC from the asymptomatic controls (0.3%). Biochemical analysis of the 20 isolates showed variability among them. However, the majority were lysine decarboxylase negative and nonmotile and utilized sodium acetate. The strains belonged to the O groups 28ac, 124, 143, or 144 or were untypable with the antisera used. Most of them had conjugative plasmids which mediated multiple antibiotic resistance. There was a strong correlation in this group of strains between a positive Sereny test, the presence of a plasmid of 120 megadaltons, and hybridization with the invasiveness probe, an HindIII fragment derived from the plasmid pPS15A. The isolates had a wide range of plasmid profiles. Bioassays and colony and Southern hybridization tests with iron uptake DNA probes indicated that 80% of the EIEC strains produced aerobactin and expressed its receptor, the genes for which are known to be chromosomally located.
Simple microbiologic detection of Campylobacter pylori
Authors: Parsonnet J , Welch K , Compton C , Strauss R , Wang T , Kelsey P , Ferraro M J
Published: January 2020

A study of 84 gastric biopsies taken from 42 adult patients revealed simple techniques for Gram stain and culture for Campylobacter pylori. In an initial study of 18 biopsies, Gram stains prepared from ground, diluted tissue were all negative for curved, gram-negative rods, whereas 13 of these specimens were positive for C. pylori by culture. The Gram stains for the remaining 66 biopsies were prepared by a rinse-imprint technique. In this group, there were 30 Gram stains positive for organisms resembling C. pylori and 32 positive cultures. By Gram-staining two sites, fundus and antrum, the sensitivity of the Gram stain for identifying a positive specimen increased from 91 to 100%. Gram stain may be the preferred technique for rapid diagnosis. When cultured, C. pylori was recovered most often on modified Thayer-Martin medium incubated microaerophilically at 35 degrees C. The presence of antibiotics in modified Thayer-Martin medium limited upper respiratory flora overgrowth, which was often present on nonselective media.
Composition and density of microflora in the subungual space of the hand
Authors: McGinley K J , Larson E L , Leyden J J
Published: January 2020

There were significant quantitative differences in the composition and density of microflora in different areas of the hands of 26 adult volunteers. The subungual spaces had an average log10 CFU of 5.39, compared with a range from 2.55 to 3.53 for other hand sites. In quantitative cultures from five subungual spaces in 26 subjects, coagulase-negative staphylococci were the dominant organisms, with Staphylococcus epidermidis, S. haemolyticus and S. hominis being the most frequently isolated species. Other bacteria recovered from subungual spaces included gram-negative bacilli in 42.3% of subjects, with Pseudomonas species composing 31.3% of this group, and coryneforms in 42.3% of subjects, with multiply resistant JK group coryneforms making up 12.5%. Yeasts were isolated from 69.0% of subjects sampled, with 51.3% of the yeasts identified as Candida parapsilosis. The subungual coagulase-negative staphylococci were susceptible to most antibiotics, with resistance to penicillin, ampicillin, and erythromycin detected in 23 to 38% of isolates.
Avidin-biotin latex agglutination assay for detection of antibodies to viral antigens
Authors: Shekarchi I C , Fuccillo D A , Sever J L , Madden D M
Published: January 2020

A new method for attaching antigens to latex by an avidin-biotin technique is described. The procedure permits control of the amount of antigen attached to the latex and eliminates the need for highly purified antigens and destructive bridging chemicals. The avidin-biotin latex agglutination assay is a simple, rapid test well suited to detection of viral antibody. The sensitivity and specificity, respectively, of the avidin-biotin latex agglutination assay compared with other assay results for antibody to viruses were as follows: cytomegalovirus, 98 and 100% (indirect hemagglutination assay); measles virus, 96 and 100% (enzyme-linked immunosorbent assay); and herpes simplex virus, 78 and 100% (indirect hemagglutination assay).
Comparative evaluation of Oxoid Signal and BACTEC radiometric blood culture systems for the detection of bacteremia and fungemia
Authors: Weinstein M P , Mirrett S , Reller L B
Published: January 2020

The Oxoid Signal (Oxoid U.S.A., Inc., Columbia, Md.) blood culture system is a newly described, innovative method for visually detecting growth of microorganisms (D. Sawney, S. Hinder, D. Swaine, and E.Y. Bridson, J. Clin. Pathol. 39:1259-1263, 1986). We did 5,999 paired comparisons of equal volumes (10 ml) of blood in the Oxoid Signal and BACTEC (Johnson Laboratories, Towson, Md.) radiometric blood culture systems at two university hospitals that use identical methods of obtaining and processing specimens. Overall, more microorganisms were detected in the BACTEC system (P less than 0.001), in particular, streptococci (P less than 0.01), fungi (P less than 0.001), and nonfermentative gram-negative rods, especially Acinetobacter species (P less than 0.001). Trends favoring the BACTEC system for detection of Pseudomonas aeruginosa, Haemophilus species, and Neisseria species were noted. There were no differences in the yield of staphylococci, members of the family Enterobacteriaceae, and anaerobic bacteria. When both systems detected sepsis, the BACTEC did so earlier (P less than 0.001). This advantage was most notable at 24 h (70% of BACTEC positives detected versus 48% of Oxoid positives). The proportion of positives detected after 48 h, however, was similar (BACTEC, 84%; Oxoid, 78%). Revisions in the Oxoid Signal system itself or in the processing of Oxoid bottles appear to be necessary to improve its performance in detecting certain microorganism groups, especially fungi.
Development of an enzyme-linked immunosorbent assay for caprine arthritis-encephalitis virus
Authors: Archambault D , East N , Perk K , Dahlberg J E
Published: January 2020

Because relatively few caprine arthritis-encephalitis virus (CAEV)-infected animals exhibit clinical signs of illness, efforts to control and eradicate this virus will depend heavily on a sensitive diagnostic test that can be easily carried out. The currently utilized tests are of limited usefulness because of relatively low sensitivity or because of incomplete cross-reactivity of goat sera with heterologous test antigens. An enzyme-linked immunosorbent assay (ELISA) with purified CAEV antigen and biotin-avidin amplification steps was therefore developed and compared with a radioimmunoassay (RIA) against CAEV p28. Of over 500 sera tested, there was 99% concordance between the two tests. On the other hand, 23 of 24 sera obtained from animals with clinical signs of disease that were negative by agar gel immunodiffusion test (with ovine progressive pneumonia virus antigen) were positive by ELISA and RIA. These results suggest that an ELISA with CAEV antigen is superior to the agar gel immunodiffusion test and is easier and faster than an RIA, and therefore may be the method of choice for diagnosing CAEV infection.
Serum and cerebrospinal fluid immunoglobulins M, A, and G in Japanese encephalitis
Authors: Han X Y , Ren Q W , Xu Z Y , Tsai T F
Published: January 2020

A comparison was made of virus-specific immunoglobulin M (IgM), IgA, and IgG detected by capture or indirect enzyme immunoassay in serum and cerebrospinal fluid of patients with Japanese encephalitis. The IgM capture enzyme immunoassay was more sensitive than assays for other isotypes of viral antibody; IgM was detected in 75% of specimens collected less than or equal to 4 days after the onset of illness. Specific IgA was detected in both serum and cerebrospinal fluid; however, IgA levels were significantly lower than IgM levels.
Rapid detection of cytomegalovirus by fluorescent monoclonal antibody staining and in situ DNA hybridization in a dram vial cell culture system
Authors: Sorbello A F , Elmendorf S L , McSharry J J , Venezia R A , Echols R M
Published: January 2020

By using dram vial cell culture methods, three commercially available tests for cytomegalovirus (CMV) detection were compared: direct fluorescent monoclonal antibody staining for CMV-specific early and late antigens (direct FA), indirect fluorescent monoclonal antibody staining for a CMV-specific early antigen (indirect FA), and in situ DNA hybridization with a biotinylated CMV-specific DNA probe kit (DNA probe). Of those tests, only the indirect FA provided consistent, reliable virus detection within the initial 24 h postinfection for serial 10-fold dilutions of CMV AD169 (laboratory strain) and for three selected urine samples. However, when used prospectively, the indirect FA failed to detect virus within the initial 10 days postinfection in 15 of 78 consecutive specimens that were eventually positive by cell culture. Although the indirect FA was more sensitive than the direct FA or DNA probe, its utility appeared limited to specimens with high CMV concentrations. On the basis of these data, we recommend that indirect FA be reserved as an adjunct to standard cell culture for selected samples in diagnostic hospital laboratories.
Immunologic and genetic comparison of Streptococcus equi isolates from the United States and Europe
Authors: Galán J E , Timoney J F
Published: January 2020

A series of isolates of Streptococcus equi from the United States and Europe were compared by the bactericidal test, immunoblotting, DNA restrictions, and Southern hybridization analysis. All isolates tested were sensitive to the same bactericidal serum. In addition, immunoblotting revealed no differences in M proteins prepared by acid or mutanolysin extraction. Immunoblotting of acid extracts of the isolates with mucosal nasopharyngeal mucus from a convalescent horse revealed the presence of the 41,000- and 46,000-Mr polypeptide fragments of the M protein of S. equi known to be important in stimulating mucosal nasopharyngeal immune responses. DNA restriction analysis of total cell DNA digests, as well as Southern hybridizations using an S. equi M protein gene probe, did not detect any differences among these isolates. Our results, therefore, confirm the antigenic homogeneity of the M proteins of S. equi isolates and suggest that variation in this antigen is not a reason for the failure of commercial vaccines in the field. Interestingly, the protoplast M proteins of all isolates showed remarkable size homogeneity, in contrast to the size variation reported in M proteins of group A streptococci.
Suggestion to supplement Shigella flexneri classification scheme with the subserovar Shigella flexneri 4c: phenotypic characteristics of strains
Authors: Pryamukhina N S , Khomenko N A
Published: January 2020

A new serovar of Shigella flexneri has been isolated with increasing frequency in the USSR. It has the antigenic formula IV:7,8. We suggest that it be incorporated into the classification scheme as S. flexneri subserovar 4c. The 212 strains of this proposed subserovar examined to date display biochemical and serological properties typical of the species, are sensitive to the genus-specific bacteriophage, and cause keratoconjunctivitis in the Sereny test.
Evaluation of an immunofluorescent-antibody test for rapid identification of Pseudomonas aeruginosa in blood cultures
Authors: Counts G W , Schwartz R W , Ulness B K , Hamilton D J , Rosok M J , Cunningham M D , Tam M R , Darveau R P
Published: January 2020

An immunofluorescent-antibody test was developed for rapid detection of Pseudomonas aeruginosa in blood cultures. The test uses a murine monoclonal antibody specific for all strains of P. aeruginosa. In initial tests, bright uniform immunofluorescence signals were seen when each of the 17 international serotypes, as well as 14 additional isolates of P. aeruginosa, were examined. No immunofluorescent staining was observed when 37 other gram-negative and 15 gram-positive species were studied. In a clinical study, the assay was applied to broth smears of 86 gram-negative bacilli isolated from 74 bacteremic patients and 28 additional clinical isolates of Pseudomonas sp. and other oxidase-positive gram-negative bacilli recovered from various body sites. Smears were made directly from blood cultures which were positive for gram-negative bacilli by Gram staining. Eleven (15%) of 74 patients with gram-negative bacteremia had a positive test for P. aeruginosa. Including the results of these 11 isolates recovered in a prospective study and an additional 10 isolates from a retrospective study, we obtained a sensitivity and specificity of 100% (21 positive specimens and 103 negative specimens, respectively). These preliminary results suggest that this is a useful reagent for rapid presumptive identification of P. aeruginosa in blood cultures. With the immunofluorescent-antibody test, P. aeruginosa could be identified within 1 h of Gram stain evidence of gram-negative bacteremia.
Occurrence of respiratory syncytial virus subgroup A and B strains in Japan, 1980 to 1987
Authors: Tsutsumi H , Onuma M , Suga K , Honjo T , Chiba Y , Chiba S , Ogra P L
Published: January 2020

The subgroup characteristics of 71 strains of respiratory syncytial virus (RSV) isolated in Sapporo, Japan, during 5 epidemic years from 1980 to 1987 were determined by the use of 17 monoclonal antibodies (MAbs) raised against the RSV Long strain, which is now recognized as the prototype subgroup A strain. Nine of these MAbs immunoprecipitated the fusion protein (F), five immunoprecipitated the large glycoprotein (G), two immunoprecipitated the nucleoprotein (NP), and one immunoprecipitated the phosphoprotein (P). Based on the pattern of reaction of these MAbs to RSV isolates in an indirect immunofluorescence assay, we were able to distinguish two different subgroups. Subgroup A strains reacted to all 17 MAbs. Subgroup B strains reacted to none of the anti-G MAbs, eight of the nine anti-F MAbs, and all anti-NP and anti-P MAbs. Subgroup A included 38 (53.5%) isolates from every epidemic year. Subgroup B included 32 (45.1%) strains isolated in the last 4 epidemic years. One virus strain with an intermediate character of reactivity was isolated in 1983. From the first epidemic year, six subgroup A strains and no B strains were isolated. During the next three annual epidemics, subgroup B strains were predominantly isolated, i.e., 8 of 13, 10 of 13, and 13 of 17 strains. However, in the last epidemic year only one strain of 22 isolates belonged to subgroup B, and the remainder belonged to subgroup A. This variability of dominance in the occurrence of different RSV subgroup strains may suggest a possible role of the subgroup-specific immune response in RSV epidemics.
Evaluation of seven immunoassays for detection of rotavirus in pediatric stool samples
Authors: Thomas E E , Puterman M L , Kawano E , Curran M
Published: January 2020

The performance of seven commercially manufactured rotavirus assays was evaluated with 144 pediatric stool specimens and compared with electron microscopy (EM) findings. The four enzyme-linked immunosorbent assays used were Rotazyme II, Pathfinder, IDL rotavirus immunoassay, and Enzygnost (Behring) rotavirus assay. The three latex tests were Meritec rotavirus detection test, Virogen Rotatest, and Bartels rotavirus latex test. Test outcomes were compared with EM on the basis of sensitivity, specificity, positive-negative predictive value, and the kappa statistic. Relative to EM, Meritec had the highest specificity (97%), followed by Virogen (95%), IDL (91%), Pathfinder (85%), Behring (81%), Bartels (72%), and Rotazyme (71%). The sensitivities were as follows: Rotazyme (92%), Pathfinder (89%), Bartels (86%), Virogen (86%), Behring (82%), Meritec (71%), and IDL (75%). Patient age and sex did not influence test results. Owing to the absence of a true standard, the tests were also compared with each other on the basis of the kappa statistic, the frequency of positive test results, and the frequency of samples in which a test differed from all other tests. Using these measures, the assays could be classified into three groups with progressively decreasing utility: group 1 (Virogen, Meritec, IDL, and EM), group 2 (Pathfinder and Behring), and group 3 (Rotazyme and Bartels). Laboratory criteria were also compared. Latex tests were faster and required less equipment than enzyme-linked immunosorbent assays. The Virogen latex assay showed the best overall performance, which made it our choice for rapid and accurate rotavirus diagnosis. However, in children who have gastrointestinal symptoms with negative rotavirus test results, EM will be useful until such time as immunological tests for other enteric viruses are available.
Distinction of species and strains of mycoplasmas (mollicutes) by genomic DNA fingerprints with an rRNA gene probe
Authors: Yogev D , Halachmi D , Kenny G E , Razin S
Published: January 2020

Genomic fingerprints of Acholeplasma laidlawii, Mycoplasma hominis, and Mycoplasma pneumoniae strains were obtained by Southern blot hybridization of the digested mycoplasmal DNAs with an rRNA gene probe. The hybridization patterns revealed genotypic heterogeneity among A. laidlawii and M. hominis strains and a remarkable degree of homogeneity among M. pneumoniae strains isolated from pneumonia patients during a 10-year period. Genomic fingerprints with the rRNA gene probe can thus serve as indicators of intraspecies genetic homogeneity or heterogeneity and can provide a new, sensitive tool for strain identification with a potential for application in epidemiology.
Relationship of health-related variables to levels of anti-polyribosylribitol phosphate antibodies in adults
Authors: McDonnell W M , Gilsdorf J R , Kent J B , Sheagren J N
Published: January 2020

To identify adults who may be at risk for Haemophilus influenzae type b disease by virtue of low levels of antibody against the H. influenzae b capsule and who may thus benefit from receiving H. influenzae b vaccine, we correlated serum anticapsular antibody levels of 388 adult patients with 26 health-related variables, including 3 personal characteristics, 10 laboratory values, 4 drug use categories, and 8 disease categories. Steroid use was consistently associated with low levels of anticapsular antibody; associations were also found between low antibody levels and both non-Caucasian race and increasing age. However, less than 4% of the antibody variability could be attributed to these factors, and they were not predictive of low antibody levels. Thus, although H. influenzae b infections are seen in adults with predisposing medical conditions, on the basis of the present findings, use of the H. influenzae b vaccine in adults cannot be recommended.
T-cell subpopulations, expression of interleukin-2 receptor, and production of interleukin-2 and gamma interferon in human American cutaneous leishmaniasis
Authors: Castes M , Cabrera M , Trujillo D , Convit J
Published: January 2020

Leukocyte subpopulations, the expression of the interleukin-2 (IL-2) receptor, and the production of IL-2 and gamma interferon (IFN-gamma) were studied in the peripheral blood mononuclear cells of American cutaneous leishmaniasis patients that had been stimulated in vitro with either leishmanial antigen or mitogen (phytohemagglutinin M). The 75 patients examined were classified as having either the localized (LCL; 66 patients), mucocutaneous (MCL; 5 patients), or the rare diffuse (DCL; 4 patients) form of the disease. Patients with DCL, who are characterized by their defective cell-mediated immune response to leishmanial antigen, failed to express the IL-2 receptor and did not produce IFN-gamma when exposed to the antigen but did so when stimulated by phytohemagglutinin M. Both LCL and MCL patients showed strong proliferative responses to leishmanial antigen; these were by far the greatest in MCL patients. Both groups had significantly increased IL-2 receptor expression and IFN-gamma production after exposure to either antigen or mitogen, and these were highest in the MCL patients. Concerning the leukocyte subpopulations evaluated (CD2, CD4, CD8, CD20, MO2), the most significant findings were a decrease of both CD4+ cells and the CD4/CD8 ratio in MCL patients compared with the other groups. Considering IL-2 production, in response to phytohemagglutinin M both MCL and LCL patients showed amounts of IL-2 comparable to those of the controls. Our results help explain the anergy of T cells from DCL patients to leishmanial antigen, which could lead to a defective production of IFN-gamma and possibly contribute to their incapacity to kill the Leishmania parasite. Concerning MCL patients, the significantly increased expression of IL-2 receptor, decreased expression of the CD4 (helper-inducer of suppression) phenotype, and elevated IFV-gamma production might partially explains the state of hypersensitivity and mucosal damage exhibited by these patients.
Postoperative wound infection associated with Vibrio parahaemolyticus in a patient without exposure to seawater
Authors: Ahsan N , Conter R L , Appelbaum P C
Published: January 2020

This report describes a case of wound infection associated with Vibrio parahaemolyticus. The patient had ingested steamed crabs 7 days before admission for surgical treatment of intestinal obstruction due to colon carcinoma. The Vibrio sp. was isolated from postoperative wound drainage as well as from stool. Recovery was uneventful.
Modified Thayer-Martin medium for recovery of Nocardia species from contaminated specimens
Authors: Murray P R , Niles A C , Heeren R L
Published: January 2020

Specimens submitted for isolation of Nocardia species are frequently contaminated with other bacteria. Although decontamination with sodium hydroxide or benzalkonium chloride can remove these bacteria, this treatment is also toxic for Nocardia species. We demonstrate that modified Thayer-Martin medium can be used for selective isolation of Nocardia species.
Comparison of three methods for recovery of Mycobacterium avium complex from blood specimens
Authors: Salfinger M , Stool E W , Piot D , Heifets L
Published: January 2020

Three methods were used for the recovery of mycobacteria from blood specimens obtained from acquired immunodeficiency syndrome patients: (i) inoculation of 7H11 agar plates with a concentrated specimen, (ii) inoculation of 7H12 BACTEC vials with a concentrated specimen, and (iii) inoculation of 7H13 BACTEC vials with a nonconcentrated specimen. In this study, we examined 255 specimens and obtained positive mycobacterial growth in 47 of them. Among these 47 cultures, 40 were found to be positive by all three methods, and the total recovery rates in relation to these culture-positive specimens were 94% for method 1, 89% for method 2, and 96% for method 3. The advantages and disadvantages of these three methods are discussed.
Chromatofocusing in the purification of staphylococcal enterotoxin D
Authors: Lei Z , Reiser R F , Bergdoll M S
Published: January 2020

A chromatofocusing procedure for the purification of staphylococcal enterotoxin D was developed. The purification included the removal of the toxic protein from culture supernatant fluids of Staphylococcus aureus 1151m by batch adsorption with CG-50 resin, chromatofocusing on Polybuffer Exchanger 94, and gel permeation chromatography on Sephacryl S-200. The purity of the staphylococcal enterotoxin D obtained was approximately 98%.
Detection of human rotaviruses which do not react with subgroup I- and II-specific monoclonal antibodies
Authors: Svensson L , Grahnquist L , Pettersson C A , Grandien M , Stintzing G , Greenberg H B
Published: January 2020

Of 126 rotavirus-positive specimens, 7 could not be subgrouped (I or II). These strains showed a distinct reaction with a monoclonal antibody recognizing a common region on VP6, but they did not react with VP6 subgroup-specific monoclonal antibodies although they contained as much viral antigen as the subgrouped strains.
Serratia odorifera biogroup 1 causing an invasive human infection
Authors: Chmel H
Published: January 2020

Serratia odorifera biogroup 1 was isolated from the blood and urine of an alcoholic male with cirrhosis and signs of septic shock. The organism is rarely reported to occur in clinical specimens. This is the first case in which the organism was found to be responsible for invasive human infection.
Synthesis of viral proteins in polymorphonuclear leukocytes infected with influenza A virus
Authors: Cassidy L F , Lyles D S , Abramson J S
Published: January 2020

Various reports have indicated that infection of polymorphonuclear leukocytes (PMNL) with influenza virus causes depression of their metabolic and chemotactic responses, but the effect the PMNL has on the life cycle of influenza virus has not been well defined. The studies reported here were undertaken to determine whether influenza virus could replicate within PMNL. Virus-infected and uninfected PMNL were labeled with [35S]methionine and analyzed by gel electrophoresis and fluorography for detection of newly synthesized proteins. Both host- and virus-specific proteins were produced within PMNL. By using indirect immunofluorescence techniques combined with flow cytometry, the expression of newly synthesized viral antigens was detected in virus-infected PMNL. Plaque assays on supernatant fluid from infected PMNL showed that infectious progeny were not produced, indicating that influenza virus infection of PMNL is abortive.
Nucleic acid hybridization studies of mycobactin-dependent mycobacteria
Authors: Yoshimura H H , Graham D Y
Published: January 2020

Using molecular techniques, specifically, DNA-DNA hybridization in solution and measurement of the thermal stabilities of hybrids, we examined the genetic relationships among mycobactin-dependent mycobacteria and between such organisms and other (mycobactin-independent) mycobacteria. The mycobactin-dependent mycobacteria examined included five strains of Mycobacterium paratuberculosis, two wood pigeon isolates, and one bovine isolate that was biochemically identified as Mycobacterium avium. All mycobactin-dependent mycobacteria were found to belong to the same hybridization group and to be closely related to organisms in M. avium complex serovars 2, 8, and 9, but more distantly related to organisms in serovars 7 and 19. Relatively low levels of hybridization were observed with other mycobacterial species examined. Thus, these results provide genetic evidence to support previous biochemical and cultural evidence that indicated that mycobactin-dependent mycobacteria constitute a closely related group of organisms within the M. avium complex.
Immunological detection of cytomegalovirus early antigen on monolayers inoculated with urine specimens by centrifugation and cultured for 6 days as alternative to conventional virus isolation
Authors: Janssen H P , van Loon A M , Meddens M J , Eickmans-Josten E C , Hoitsma A J , de Witte T J , Quint W G
Published: January 2020

Detection methods for human cytomegalovirus were evaluated with 431 urine samples from 30 bone marrow and 88 kidney transplant recipients. Low-speed centrifugal inoculation was followed by early antigen (EA) detection by means of indirect immunofluorescence with a monoclonal antibody after 1 (EA-1) and 6 (EA-6) days of cultivation. The results were compared with those of conventional virus isolation (CVI). Of 68 positive samples, 49 (72%) were detected with EA-1, 58 (85%) were detected with EA-6, and 43 (63%) were detected with CVI. The combination of EA-1 and EA-6 showed positive results with 66 samples (97%), which is significantly better than with CVI (P less than 0.001). With the exception of one patient, all CVI-negative but EA-positive samples had either significant rises in immunoglobulin G (IgG) or IgA antibody titer or IgM antibodies present in the sera. These data indicate that the method with EA detection can replace CVI, provided that each sample is inoculated in duplicate. Sample 1 is examined after 1 day, and if it is negative, sample 2 is incubated for a further 5 days, followed by detection of cytomegalovirus.
Kinetics of delta antigen and delta antibody in acute delta hepatitis: evaluation with different enzyme immunoassays
Authors: Dubois F , Goudeau A
Published: January 2020

The kinetics of delta antigen (HDAg) and anti-delta antibody (anti-HD) was analyzed in 22 acute delta hepatitis infections (11 coinfections and 11 superinfections), with an enzyme immunoassay developed by Organon Teknika and with two commercially available assays: Deltassay, a test for HDAg from Noctech and Abbott anti-delta enzyme immunoassay, a test for anti-HD from Abbott Laboratories. In seven cases, HDAg was detected with the Organon assay but not with Deltassay. These discrepancies were not related to the type of hepatitis delta virus infection. All samples from these patients taken beyond week 4 of illness were found anti-HD positive with both the Organon and Abbott anti-HD assays. These data reflect the lack of sensitivity of Deltassay. In no instance were HDAg and anti-HD present simultaneously when tested with the Organon assays. On the contrary, 10 sera among the 28 that were HDAg positive with the Organon assay also were found anti-HD positive with the Abbott test. We suspected that the test procedure recommended by Abbott (a one-step competitive assay) may have yielded false-positive anti-HD results when HDAg present in the sera reacted with peroxidase-labeled anti-HD of the kit. To determine the specificity of the simultaneous presence of HDAg and anti-HD, these 10 sera were retested with the Abbott anti-HD assay, but by a modified two-step procedure that avoided contact between sera and labeled antibody. For six sera a negative result was obtained with the second procedure, suggesting that a false anti-HD reaction occurred with the standard test procedure. For four sera a positive result with both procedures was indicative of the simultaneous presence of HDAg and anti-HD. In conclusion, assay for HDAg was found very convenient for the early diagnosis of acute hepatitis delta virus infections. Seroconversion to anti-HD could be used for a late diagnosis 2 to 5 weeks after the beginning of illness. However, anti-HD results obtained with one-step competitive assays have to be interpreted carefully in HDAg-positive sera.
Characterization of flagella purified from enterohemorrhagic, vero-cytotoxin-producing Escherichia coli serotype O157:H7
Authors: Sherman P , Soni R , Yeger H
Published: January 2020

Escherichia coli of the serotype O157:H7 has recently been isolated in human fecal specimens in association with sporadic cases and outbreaks of hemorrhagic colitis and with the hemolytic uremic syndrome. The aim of this study was to characterize the flagellin protein subunit constituents of flagellar filaments from E. coli O157:H7 strain CL-56. Flagellin isolated from a reference Salmonella enteritidis strain was used for comparison. Flagella were dissociated by incubation of bacteria under acidic conditions, centrifugation, and differential ammonium sulfate precipitation. Reconstituted flagellar filaments were demonstrated by three complementary methods: transmission electron microscopy, antigenic reactivity with H7 antiserum by a dot blot immunoassay, and immunogold localization of antiserum raised to the purified antigen to intact flagella on whole E. coli O157:H7. On sodium dodecyl sulfate-polyacrylamide gels flagellin proteins from E. coli O157:H7 demonstrated an apparent Mr of 66,000. The isoelectric point of E. coli O157:H7 flagellin was 5.42. By immunoblotting, H7 flagellin proteins were shown to be immunogenic. They induced a systemic immune response both in rabbits challenged with whole bacteria and in a human previously infected with E. coli O157:H7.
Immunoblot analysis of humoral immune responses following infection with Bordetella pertussis or immunization with diphtheria-tetanus-pertussis vaccine
Authors: Redd S C , Rumschlag H S , Biellik R J , Sanden G N , Reimer C B , Cohen M L
Published: January 2020

To help develop better diagnostic tests for pertussis, we examined the serologic response to whole-cell proteins of Bordetella pertussis after natural infection or vaccination with diphtheria-tetanus-pertussis vaccine. Serum specimens collected during a pertussis outbreak investigation and from uninfected persons were used in Western blot (immunoblot) analyses to determine the presence of immunoglobulin G (IgG) and IgA antibodies to specific B. pertussis proteins. IgG antibodies to proteins of molecular masses 220 and 210 kilodaltons (kDa) were detected in 14 of 18 serum samples obtained from patients with culture-confirmed pertussis greater than or equal to 40 days after the onset of coughing. IgA antibodies were detected in 15 of the 18 samples. Of 19 serum samples obtained from patients who had not been ill with pertussis, 6 contained IgG antibodies to these proteins and 1 contained IgA antibodies. The two proteins bound antiserum specific for filamentous hemagglutinin and comigrated with purified filamentous hemagglutinin. IgG antibodies to two additional protein bands of molecular masses 84 and 75 kDa were associated with previous vaccination. Antibody to the 84-kDa protein was detected in 15 of 17 vaccinated, never-infected persons, and antibody to the 75-kDa protein was detected in 16 of the 17. None of 11 nonvaccinated, never-infected persons tested had antibodies to either protein. All seven fully vaccinated persons with culture-documented infection had antibodies to both proteins. Antibodies to the 84-kDa protein were detected in 6 of 22 nonvaccinated and infected persons, and antibodies to the 75-kDa protein were detected in 8 of the 22. Use of Western blot analysis in this study allowed us to distinguish antibody responses to infection and immunization.
Rapid method for the differentiation of gram-positive and gram-negative bacteria on membrane filters
Authors: Romero S , Schell R F , Pennell D R
Published: January 2020

Microfiltration has become a popular procedure for the concentration and enumeration of bacteria. We developed a rapid and sensitive method for the differentiation of gram-positive and gram-negative bacteria, utilizing a polycarbonate membrane filter, crystal violet, iodine, 95% ethanol, and 6% carbol fuchsin, that can be completed in 60 to 90 s. Gram reactions of 49 species belonging to 30 genera of bacteria were correctly determined by the filter-Gram stain. The sensitivities of the filter-Gram stain and conventional slide-Gram stain were compared by testing dilutions of Escherichia coli, Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae suspensions in the presence and absence of whole human blood. The filter-Gram stain was approximately 100-fold more sensitive than the slide-Gram stain. The filter-Gram stain detected 2 to 100 bacteria, whereas the slide-Gram stain failed to detect less than 1,000 bacteria. The sensitivities of the methods were not significantly altered by the addition of whole human blood to the dilutions of bacteria tested. The filter-Gram stain could be a useful tool for the examination of body fluids with very low numbers of bacteria.
Comparative sensitivities of solid-phase immune electron microscopy and enzyme-linked immunosorbent assay for serotyping of human rotavirus strains with neutralizing monoclonal antibodies
Authors: Gerna G , Sarasini A , Coulson B S , Parea M , Torsellini M , Arbustini E , Battaglia M
Published: January 2020

Suspensions of 24 rotavirus strains, 6 for each known human rotavirus serotype, were serially diluted and titrated by (i) enzyme-linked immunosorbent assay (ELISA) for rotavirus detection, using monoclonal antibodies (MAbs) specific for group-specific sites of the VP6 inner capsid protein; (ii) ELISA for subgrouping, using MAbs reactive with subgroup-specific determinants of rotavirus VP6; (iii) ELISA for serotyping, using MAbs directed to serotype-specific sites of the VP7 outer capsid glycoprotein; and (iv) solid-phase immune electron microscopy (SPIEM) for serotyping, using VP7-specific MAbs. In addition, in each preparation the proportion of double-shelled rotavirus particles were determined by direct electron microscopy. Results showed that SPIEM was 2- to 16-fold more sensitive than ELISA for serotyping of rotavirus. The titers in VP7-specific tests correlated well with the proportion of double-shelled virus particles in each of the samples. Titers obtained by ELISA for serotyping of suspensions containing 20% or fewer complete particles were up to 4,096-fold lower than those obtained by ELISA for detection. ELISA serotyping titers of samples containing 20 to 80% double-shelled rotavirus particles were up to 128-fold lower than ELISA detection titers, whereas preparations with nearly 100% complete particles had ELISA titers that were less different from each other. ELISA subgrouping titers were four- to eightfold lower than corresponding rotavirus detection titers. It was concluded that, although SPIEM appears to be more sensitive than ELISA, the amount of complete virus particles in the specimens is of critical importance for successful serotyping of human rotavirus strains. Samples rich in single-shelled particles but containing low amounts of VP7 outer capsid glycoprotein might even be strongly reactive in assays for rotavirus detection and subgrouping but virtually unreactive in tests for serotyping.
Identification of two subtypes of serotype 4 human rotavirus by using VP7-specific neutralizing monoclonal antibodies
Authors: Gerna G , Sarasini A , di Matteo A , Parea M , Orsolini P , Battaglia M
Published: January 2020

Two distinct subtypes of human rotavirus serotype 4 were identified by using neutralizing monoclonal antibodies directed to the major outer capsid glycoprotein, VP7, of strains ST3 (subtype 4A) and VA70 (subtype 4B). Specimens containing serotype 4 rotavirus, obtained from different countries, were examined for subtyping by using solid-phase immune electron microscopy, enzyme-linked immunosorbent assay, and, for cell culture-adapted strains, neutralization assay. All 59 human rotavirus strains identified as serotype 4 by using animal antisera were classified into either subtype by monoclonal antibodies. This suggests that the antigenic difference between the two subtypes is a consequence of critical variations within the immunodominant serotype 4-specific neutralization site of rotavirus VP7. Subtype 4A (ST3-like) strains were predominant and were detected in stools from patients with gastroenteritis, as well as from healthy infants and young children.
Effect of egg yolk on growth of Mycobacterium tuberculosis in 7H12 liquid medium
Authors: Kononov Y , Ta K D , Heifets L
Published: January 2020

Of 92 drug-resistant Mycobacterium tuberculosis strains isolated from sputum specimens, 86 showed growth in two types of 7H12 broth, one with egg yolk and the other without egg yolk. In addition, two strains grew only in plain 7H12 broth without yolk, and four others were recovered only in the medium supplemented with egg yolk. The radiometrically detected growth was higher in the presence of egg yolk, corresponding to a higher number of CFU per milliliter in these cultures. The improvement of growth in 7H12 broth supplemented with egg yolk was most noticeable in cultures isolated from sputum specimens having a low number of acid-fast bacilli in the smear and producing only a few colonies on solid media.