bluuebunny/crossref_metadata_2025 huggingface dataset:
Diagnosis of pulmonary tuberculosis by selected-ion monitoring: improved analysis of tuberculostearate in sputum using negative-ion mass spectrometry
Authors: Larsson L , Odham G , Westerdahl G , Olsson B
Published: January 2020

Coded sputum specimens from patients with tuberculous (8 specimens) and nontuberculous (15 specimens) pneumonia were analyzed for tuberculostearic acid by selected-ion monitoring of the pentafluorobenzyl ester derivative by using negative-ion mass spectrometry. The analytical results agreed with those from culturing of the specimens, except in three cases in which the chromatograms were difficult to interpret owing to insufficient amounts of sputa available. Negative-ion mass spectrometry provided superior sensitivity and selectivity for detection of tuberculostearic acid compared with the previously used gas chromatographic-mass spectrometric methods. The technique reported here should have considerable potential for the rapid diagnosis of pulmonary tuberculosis by direct analysis of sputum.
Rapid diagnosis of human Crimean-Congo hemorrhagic fever and detection of the virus in naturally infected ticks
Authors: Saluzzo J F , Le Guenno B
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) was developed to detect Crimean-Congo hemorrhagic fever (CCHF) virus-specific immunoglobulin M (IgM) in human serum samples. For this test, a heat-inactivated antigen was prepared from the brains of suckling mice infected with CCHF virus. The IgM-capture ELISA proved more sensitive than indirect fluorescence tests for IgM to this virus. A human serum containing high-titer IgM to CCHF virus was used for an antigen-capture ELISA to detect this virus in heat-inactivated suspensions of virus-infected ticks. The antigen-capture ELISA appeared to be as sensitive as virus isolation in suckling mice. The studies described suggest that the IgM-capture ELISA and the antigen-detection ELISA should provide a rapid and sensitive diagnosis of human CCHF virus infection and should be useful in ecologic studies of this virus.
Sodium chloride as aid in identification of Phaeoannellomyces werneckii and other medically important dematiaceous fungi
Authors: Kane J , Summerbell R C
Published: January 2020

Seventeen taxa of dematiaceous fungi isolated from humans were tested to determine their responses to various concentrations of sodium chloride in vitro. Five groups of species were recognized on the basis of differing tolerances. Phaeoannellomyces werneckii was distinguished by its tolerance of greater than or equal to 15% NaCl; most dematiaceous pathogens were suppressed at less than or equal to 7% NaCl.
Economical laboratory support system for influenza virus surveillance
Authors: Reichelderfer P S , Kappus K D , Kendal A P
Published: January 2020

When increases in influenzalike illnesses were first detected during the 1984 to 1985 and 1985 to 1986 influenza seasons, throat swab specimens were mailed by physicians across the United States to one hospital laboratory, and each specimen was inoculated into a single tube of cell culture. Of 165 specimens, 52 (32%) were positive for influenza A or B virus within 3 days of receipt when tested by fluorescence microscopy, permitting positive results to be received by the physicians within 1 week on the average. Approximate direct cost was $16 per specimen.
Isolation of group A swine rotaviruses displaying atypical electropherotypes
Authors: Bellinzoni R C , Mattion N M , Burrone O , Gonzalez A , La Torre J L , Scodeller E A
Published: January 2020

Swine rotaviruses displaying distinctive electropherotypes were isolated from the feces of diarrheic piglets in two swine herds in the province of Buenos Aires, Argentina. In one case all samples isolated showed abnormal electropherotypes. All samples were classified as group A reactive when assayed by an enzyme-linked immunosorbent assay. Three samples from this herd were adapted to grow in tissue culture. The electrophoretic pattern of the genomic RNA as well as the group A reactivity of one of these viruses was retained after cloning in MA-104 cells. In the other pig unit were found samples displaying both classical and abnormal electropherotypes. These viruses were also positive in the enzyme-linked immunosorbent assay; however, since they could not be adapted to grow in tissue culture, this classification must be considered tentative. The abnormal electropherotype exhibited by these pig viruses strongly resembles those of human origin called super short.
Development of immunity in natural Plasmodium falciparum malaria: antibodies to the falciparum sporozoite vaccine 1 antigen (R32tet32)
Authors: Webster H K , Boudreau E F , Pang L W , Permpanich B , Sookto P , Wirtz R A
Published: January 2020

Antibodies that reacted with a candidate sporozoite vaccine antigen (R32tet32) were found in 20 of 21 patients treated for acute infection with Plasmodium falciparum and monitored longitudinally over 67 days. R32tet32 contains 32 tandem copies of a tetrapeptide sequence that constitutes the immunodominant epitope of the circumsporozoite surface protein. The magnitude of the antibody response varied considerably among individuals and appeared to be independent of the number of previous clinical infections. Recrudescence of infection or infection with Plasmodium vivax had no demonstrable effect on antibody levels, although reinfection with P. falciparum produced a rapid rise in antibody titer. Antibody levels were observed to decline rapidly after treatment of clinical infection with mefloquine. The apparent antibody half-life was 27 days, which is comparable to the half-life of circulating immunoglobulin G in humans. The data suggest that antisporozoite antibody production ceased on about day 4 after treatment of acute infection. A similar pattern of response was observed for antibodies against the erythrocytic forms of the parasite. The cessation of antibody synthesis was interpreted as being due to immunosuppression induced by the presence of intraerythrocytic parasites and may explain in part why protective immunity is poorly developed in natural malaria infections.
Development and evaluation of a capture enzyme-linked immunosorbent assay for determination of rubella immunoglobulin M using monoclonal antibodies
Authors: Gerna I , Zannino M , Revello M G , Petruzzelli E , Dovis M
Published: January 2020

A capture enzyme-linked immunosorbent assay (ELISA) for detection of virus-specific immunoglobulin M (IgM) antibody was developed which used a panel of labeled monoclonal antibodies to rubella virus hemagglutinin. The rapidity of the test system was increased by using, after 1-h incubation of the test serum, a second 1-h incubation of the serum with a mixture of viral antigen and labeled monoclonal antibody. The new assay was tested for specificity on 371 human sera from people without any recent contact with rubella virus; of these, 66 were sera selected from people with rheumatoid factor or IgM antibody to human cytomegalovirus, Epstein-Barr virus, or other viruses. In parallel, the new assay was performed on 191 sera from patients having recent contact with rubella virus. Results were compared with those obtained by an indirect ELISA method on IgM serum fractions, using purified rubella virus as a solid phase. Of the 371 sera tested for specificity, 5 (1.3%) gave false-positive results with indirect ELISA (1 rheumatoid factor, 2 heterophil antibody, and 2 human cytomegalovirus sera positive for IgM), and none were false-positive with the capture assay. Two sera from a patient with primary cytomegalovirus infection, which were positive for rubella IgM antibody with both methods and were initially interpreted as false-positive, were finally considered to be true-positive, since they were reactive only in the presence of IgM antibody and viral antigen. Of the 191 sera from 92 patients (84 patients with acute rubella, four newborns from mothers with rubella during pregnancy, and four vaccinees), 136 (71.2%) were found to be positive for IgM by direct ELISA, and 128 (67.0%) were positive by capture ELISA; 12 sera drawn during the first 2 days of disease, or at least 40 days after onset (or after vaccination), were detected only by indirect ELISA, and 4 sera were detected only by capture ELISA. Thus, specificity and sensitivity, respectively, were 100 and 91.4% for capture ELISA and 98.6 and 97.1% for indirect ELISA. However, when the number of patients was considered, 86 were detected as IgM positive by indirect ELISA, and 87 were detected positive by capture ELISA. The overall agreement between the two assays was 96.2%. Capture ELISA using monoclonal antibody appears preferable over indirect ELISA on IgM serum fractions because of its higher specificity and shorter time for test performance; furthermore, there is no need for serum fractionation or virus purification for the capture ELISA.
Multiplicity of serogroups and adhesins in enteropathogenic and enterotoxigenic Escherichia coli isolated from acute diarrhea in Senegal
Authors: Darfeuille-Michaud A , Forestier C , Masseboeuf R , Rich C , M'Boup S , Joly B , Denis F
Published: January 2020

Escherichia coli strains were isolated from 228 children with diarrhea in Senegal from 1982 to 1984. Among these E. coli involved in cases of diarrhea, we found that 20.3% were enteropathogenic E. coli. Only 3.9% of the strains adhered to the brush borders of human intestinal enterocytes, and they belonged to different serotypes. All these adhesion-positive strains possessed genes encoding for the heat-stable enterotoxin, but their adhesive factors were different regarding serology with anti-colonization factor sera, hemagglutination patterns, electron microscopy structures, or major surface protein subunits.
Differentiation of Bordetella avium and related species by cellular fatty acid analysis
Authors: Moore C J , Mawhinney H , Blackall P J
Published: January 2020

The fatty acids of 18 strains of Bordetella avium, 3 strains of Alcaligenes faecalis, 5 strains of Bordetella bronchiseptica, and 12 strains of a B. avium-like organism were examined by gas chromatography-mass spectrometry. The presence of a significant amount of the acid 2-OH C14:0 characterized B. avium and the B. avium-like organism. B. avium and the B. avium-like organism differed in their relative concentrations of C16:1 and 3-OH C14:0 acids. B. bronchiseptica and A. faecalis were distinguishable by comparison of the relative concentrations of C18:0 and C18:1 acids.
New selective agar medium for isolation of virulent Yersinia enterocolitica
Authors: Fukushima H
Published: January 2020

A selective agar medium for isolation of virulent Yersinia enterocolitica (VYE agar) was developed for the rapid and accurate isolation of virulent Y. enterocolitica from environmental samples highly contaminated with environmental Yersinia organisms, as well as for isolation from clinical specimens. VYE agar provided a quantitative recovery of 51 different strains of virulent Y. enterocolitica at 32 degrees C after incubation for 24 h. The cefsulodin, irgasan, josamycin, and oleandomycin content of the medium resulted in a high selectivity, and the mannitol and esculin content provided some differentiation. The greatest advantage of VYE agar is that virulent Y. enterocolitica, which forms red colonies, is easily differentiated from most environmental Yersinia organisms and other gram-negative bacteria, which form dark colonies with a dark peripheral zone as a result of esculin hydrolysis. Use of VYE agar led to a high recovery of Y. enterocolitica biotype 3B serotype O:3 strains from experimentally inoculated meat samples, compared with use of CIN agar. Biotype 2 serotypes O:5,27 and O:9 and biotype 1 esculin-negative serotypes O:4,32, O:8, O:13a,13b, O:18, O:20, and O:21 (American types) were readily differentiated from other environmental organisms able to grow on VYE agar. Epidemiological studies on Y. enterocolitica should be greatly facilitated by the use of this selective agar medium.
Staphylococcal adherence to polyvinyl chloride and heparin-bonded polyurethane catheters is species dependent and enhanced by fibronectin
Authors: Russell P B , Kline J , Yoder M C , Polin R A
Published: January 2020

Intravenous hyperalimentation has improved the survival of premature infants. However, long-term placement of intravenous catheters may result in the development of catheter-related sepsis. Fibronectin in plasma contains binding sites for staphylococcal species as well as marked affinity for inert plastics and therefore may provide a substrate for bacterial adherence to indwelling catheters. We determined the adherence of labeled [( 3H]leucine) coagulase-positive (CPS) and coagulase-negative (CNS) staphylococci to untreated and fibronectin-coated polyvinyl chloride (PVC) and heparin-bonded polyurethane (HBP) catheter segments and quantitated the binding of 14C-labeled, purified fibronectin to these catheters. PVC catheter segments bound significantly more CNS than CPS (P less than 0.05), while HBP catheters bound more CPS than CNS (P less than 0.05). Fibronectin significantly increased the adherence of CPS to PVC catheters (P less than 0.05) and CNS to HBP catheters (P less than 0.05). PVC catheters bound more fibronectin (P less than 0.0001) than did HBP catheters. Catheter composition may influence the spectrum of nosocomial pathogens to which infants are susceptible through different bacterial adherences and interactions with adhesive proteins.
Sodium chloride-esculin hydrolysis test for rapid identification of enterococci
Authors: Qadri S M , Flournoy D J , Qadri S G
Published: January 2020

The ability of enterococci to cause severe disease in humans and their relative resistance to chemotherapeutic agents make it desirable to rapidly differentiate these organisms from other streptococci. We developed and evaluated a test that within 2 h distinguishes enterococci from other alpha-, beta-, or nonhemolytic streptococci in a buffered solution containing 0.2% esculin and 5% sodium chloride. All 239 strains of enterococci tested gave a positive reaction within 2 h, whereas 95 of 96 isolates of other streptococci remained negative at 4 h.
Comparison of rapid identification method and conventional substrates for identification of Corynebacterium group JK isolates
Authors: Grasmick A E , Bruckner D A
Published: January 2020

In an effort to identify Corynebacterium group JK isolates rapidly, Rapid Identification Method (RIM series; Austin Biological Laboratories, Inc., Austin, Tex.) substrates were tested in parallel with conventional substrates. RIM reactions agreed with conventional substrate results, respectively, as follows: urea, 38 of 38; nitrate, 35 of 38; glucose, 35 of 38; maltose, 28 of 38; sucrose, 37 of 38; and o-nitrophenyl-beta-D-galactopyranoside, 24 of 26. As a supplement to initial screening tests, the RIM tests offer a rapid method for identifying group JK isolates.
Recurrent Pseudomonas luteola (CDC group Ve-1) peritonitis in a patient undergoing continuous ambulatory peritoneal dialysis
Authors: Connor B J , Kopecky R T , Frymoyer P A , Forbes B A
Published: January 2020

Recurrent Pseudomonas luteola (CDC group Ve-1) peritonitis occurred in a patient undergoing continuous ambulatory peritoneal dialysis. Catheter removal was required for cure despite therapy based on antibiotic susceptibilities. This is the third report in the English literature of severe P. luteola infection and the first report of peritonitis caused by this organism.
Detection of human immunodeficiency virus by reverse transcriptase assay, antigen capture assay, and radioimmunoassay
Authors: Gupta P , Balachandran R , Grovit K , Webster D , Rinaldo C
Published: January 2020

The reverse transcriptase assay, antigen capture assay, and radioimmunoassay were compared for the detection of human immunodeficiency virus (HIV) in culture fluids of virus-infected lymphocytes. The reverse transcriptase assay and the antigen capture assay were compared for 962 samples, and the two tests displayed comparable sensitivities (98.5% agreement) in detecting HIV antigen. In addition, these two tests displayed similar sensitivities when examined for the kinetics of HIV appearance following in vitro infection of normal lymphocytes. We also found the antigen capture assay to be as sensitive as the radioimmunoassay in detecting HIV antigen in culture fluids. Furthermore, all three tests were found to be reasonably concordant when applied simultaneously to the detection of HIV antigen in cultures. The antigen capture assay, however, is relatively fast, can handle a large number of samples, does not require radioactive material, and is less expensive than the other two tests. Therefore, the antigen capture assay appears to be the method of choice for the detection of HIV in routine virus culturing.
Two-year survey of etiologic agents of diarrheal disease at San Lazaro Hospital, Manila, Republic of the Philippines
Authors: Adkins H J , Escamilla J , Santiago L T , Rañoa C , Echeverria P , Cross J H
Published: January 2020

The prevalence of bacterial pathogens and rotavirus in 2,908 patients with diarrhea who were admitted to San Lazaro Hospital in Manila in 1983 and 1984 was determined. One or more enteric pathogens were isolated or detected in samples from 1,698 (58.4%) patients. Isolation rates for the various enteropathogens were as follows: rotavirus, 30.6%; Shigella spp., 11.6%; Salmonella spp., 9.2%; enterotoxigenic Escherichia coli (1983 only), 7.8%; Vibrio cholerae biotype eltor, 3.8%; non-O1 V. cholerae, 2.8%; Vibrio parahaemolyticus, 1.7%; other Vibrio spp., 1.1%; Campylobacter jejuni, 3.0%; Aeromonas hydrophila, 1.3%; and Plesiomonas shigelloides 1.1%. Giardia lamblia and Entamoeba histolytica were detected in 0.6 and 0.1%, respectively, of stool samples examined. Determination of the etiologic role of isolates was complicated by one or more of the following factors: isolation of multiple enteric pathogens (302 cases); isolation of Salmonella spp., enterotoxigenic E. coli, and C. jejuni from a similar proportion of asymptomatic control patients and patients with diarrhea; and isolation of a high proportion of certain pathogens (especially Salmonella spp.) only from enrichment broth, suggesting infection with a small number of organisms. Isolation of V. cholerae eltor was seasonal, with the majority of cases occurring in the rainy months. In addition, the number of patients with diarrhea increased with the onset of the monsoon rains and peaked during the months of maximum rainfall. Rotavirus infection occurred in both children and adults throughout the year and was the most frequently identified cause of diarrhea in children under 5 years of age. Shigella spp. were the most common agents of diarrhea in adults.
Isolation of Borrelia spirochetes from patients in Texas
Authors: Rawlings J A , Fournier P V , Teltow G J
Published: January 2020

The Texas Department of Health Laboratory began culturing the Lyme disease spirochete Borrelia burgdorferi in 1985. This organism was subsequently isolated from blood, cerebrospinal fluid, joint fluid, skin, bone, and autopsy tissues from humans. Fluorescent-antibody tests with murine monoclonal antibodies confirmed that seven of these isolates were B. burgdorferi and that two others belonged to the genus Borrelia.
Antigens that are similar in apparent molecular weight to gonococcal outer membrane protein III
Authors: Barrera O , Swanson J
Published: January 2020

Two immunoglobulin G monoclonal antibodies (MAbs) were produced by using purified outer membranes and whole gonococci as immunogens. These MAbs recognized antigens with similar apparent sizes (30 to 31.5 kilodaltons [kDa]) in several pathogenic and nonpathogenic neisseriae. In gonococci, these 30- to 31.5-kDa components, although similar in subunit size to outer membrane protein III (P.III), are distinct due to their differences in electrophoretic migration-modification by 2-mercaptoethanol and their cellular location. The two 30- to 31.5-kDa moieties are denoted by MAb2 and MAb3, respectively, by which they are identified. The differentiating characteristic of these three antigens (P.III, MAb2, MAb3) is their change or lack of change in electrophoretic mobility in the presence versus absence of 2-mercaptoethanol; P.III migrates less rapidly, MAb2 does not change, and MAb3 migrates more rapidly. Both the epitopes that were reactive with MAb2 and MAb3 were resistant to proteolytic treatment of intact gonococci; neither epitope was detected on whole, unfixed gonococci by immunofluorescence. Both MAb2 and MAb3 epitopes were represented uniformly among pathogenic neisseriae (except group Z meningococci) and less regularly among nonpathogenic neisseriae.
Expression of proteins of Mycobacterium tuberculosis in Escherichia coli and potential of recombinant genes and proteins for development of diagnostic reagents
Authors: Cohen M L , Mayer L W , Rumschlag H S , Yakrus M A , Jones W D , Good R C
Published: January 2020

Recombinant plasmids containing DNA from Mycobacterium tuberculosis were transformed into Escherichia coli, and three colonies were selected by their reactivity with polyclonal antisera to M. tuberculosis. The three recombinant vectors contained DNA inserts of different sizes flanking a common 4.7-kilobase (kb) sequence. Each recombinant produced 35- and 53-kilodalton proteins (35K and 53K proteins, respectively) which were absent in the control E. coli. In Western blotting experiments, both proteins bound several antisera to M. tuberculosis but not antisera to other commonly isolated mycobacteria. Rabbits immunized with the recombinant 35K protein produced antisera which bound to both the 35K and 53K protein bands, a single 35K protein band present in a culture filtrate of M. tuberculosis, and single protein bands with differing molecular weights in whole-cell homogenates from other Mycobacterium spp. An additional recombinant vector containing a 2.2-kb subclone of the 4.7-kb sequence was constructed and, when used as a probe, demonstrated homology with various fragments of chromosomal digests of selected mycobacteria. Reactivity of this probe to Mycobacterium bovis and M. bovis BCG was indistinguishable from reactivity to M. tuberculosis. Immunoglobulin G reactivity to the 35K antigen was detected in antisera from 8 of 20 persons with active tuberculosis, 4 of 18 persons with leprosy, and none of 14 healthy controls. In contrast, reactivity to various proteins in M. tuberculosis culture filtrate was present in 18 of 20 patients with tuberculosis, 16 to 18 patients with leprosy, and 5 of 14 controls. The production of M. tuberculosis proteins by E. coli circumvents many difficulties encountered in the growth and manipulation of M. tuberculosis and may facilitate the development of better diagnostic and immunizing reagents.
Collaborative clinical laboratory study of a broth-disk test for determination of bacterial susceptibility to beta-lactams in combination with amdinocillin
Authors: Isenberg H D , Tilton R C , Barry A L , Beskid G , Cleeland R , Fallat V , Murray P R , Pierson C , Squires E
Published: January 2020

Methodology for the performance of synergistic antibiotic susceptibility studies has not been standardized. We addressed this problem collaboratively with combinations of amdinocillin and select other beta-lactam antibiotics by using a simple broth-disk test compared with a microdilution approach. Each method used the same drugs singly and in combination. The broth-disk test evaluated each agent and the combinations at concentrations that reflected the breakpoints for each drug; the same ratios of beta-lactam to amdinocillin were used in doubling dilutions with the microdilution method. Initially, each participant studied the same 50 members of the family Enterobacteriaceae; each bacterium was studied on three occasions. Thereafter, 500 representatives of Enterobacteriaceae isolated recently from clinical specimens were studied. Designated strains served as controls. Reproducibility between the two approaches studied in phase 1 of the investigation indicated good agreement between the methods, ranging from 87 to 100%. Agreement between the microdilution and broth-disk tests for the 2,551 clinical isolates ranged from 86 to 95%, with slightly better correlations between combination results than with the single agents. The findings indicate that antibiotic disks used routinely in the clinical laboratory can be used in a simple elution test to determine susceptibility of organisms to beta-lactam antibiotics alone and in combination with amdinocillin.
Effect of acidity and antimicrobial agent-like compounds on viability of Plesiomonas shigelloides
Authors: Janda J M
Published: January 2020

Nineteen Plesiomonas shigelloides strains were evaluated for their stability at acidic and slightly alkaline pHs and for their susceptibility to antimicrobial agent-like compounds produced by enteric flora. Most P. shigelloides isolates were rapidly inactivated under high-acid (pH 4 or less) conditions. Screening of enteric bacteria for elaboration of factors active against P. shigelloides revealed two organisms (Pseudomonas aeruginosa and Streptococcus [Enterococcus] faecium) capable of secreting such inhibitory substances. The results of this study suggest some factors potentially important in regulating gastrointestinal colonization by P. shigelloides from environmental sources.
New bacteriophage typing scheme for subdivision of the frequent capsular serotypes of Klebsiella spp
Authors: Gaston M A , Ayling-Smith B A , Pitt T L
Published: January 2020

A bacteriophage typing scheme for hospital isolates of Klebsiella spp. was developed. The scheme was designed specifically as a secondary typing method to discriminate between strains of serotypes K2, K3, and K21 but proved to be an efficient general typing method for strains of most serotypes. The set of 15 phages gave 87.3% typeability on 236 strains of more than 70 different serotypes. Typeability within the K2, K3, and K21 strains was 93, 89, and 91%, respectively. There was a mean of 3.2 reactions strain-1 for all phage-typeable strains. Of the serologically nontypeable strains, 76.7% were susceptible to one or more phages. The most common pattern accounted for only 7% of the strains. The lytic patterns were reproducible if strains were typed on the same day, but differences were observed if strains were stored for 1 week or more before retyping. A total of 96.5% of the strains were typeable by a combination of capsular serology and phage typing.
Atypical campylobacters associated with gastroenteritis
Authors: Tee W , Anderson B N , Ross B C , Dwyer B
Published: January 2020

Nine strains of Campylobacter species other than Campylobacter jejuni, Campylobacter coli, and Campylobacter laridis were isolated from patients with acute diarrhea. All nine strains showed preferred growth at 37 degrees C under microaerophilic conditions. Conventional microbiological tests and DNA-DNA dot blotting were used to identify these strains. Three of the nine Campylobacter strains hydrolyzed hippurate, reduced nitrate, produced catalase, were resistant to cephalothin, and were shown to be highly related to C. jejuni type strains. Two strains had negative or weak catalase activity and were hippurate negative. Three other strains had characteristics similar to those of Campylobacter cinaedi. The ninth strain, isolated from a homosexual man with antibodies to human immunodeficiency virus (human T-cell lymphotropic virus type III), showed unique features different from those of all the known campylobacters used in this study. This strain grew well at 25 and 37 degrees C and was catalase and nitrate positive, hippurate negative, and resistant to cephalothin.
Kinetics of anti-Campylobacter jejuni monomeric and polymeric immunoglobulin A1 and A2 responses in serum during acute enteritis
Authors: Mascart-Lemone F O , Duchateau J R , Oosterom J , Butzler J P , Delacroix D L
Published: January 2020

The intensity and kinetics of the serum polymeric and monomeric immunoglobulin A1 (IgA1) and IgA2 antibody responses to Campylobacter jejuni were analyzed. A rapid and marked serum IgA antibody response involving both the monomeric and polymeric components of IgA was observed after C. jejuni infections. IgA antibodies reached a peak of activity in serum during week 2 after the first symptoms of enteritis, about 10 days before the peak of IgG activity. Polymeric IgA accounted for most of the anti-C. jejuni activity at the peak of the IgA response (median, 90%; range, 44 to 98%) but rapidly disappeared from serum over a few weeks. In contrast, the serum monomeric IgA antibody response was low and was maintained over a prolonged period of time. Anti-C. jejuni IgA detected in the serum of healthy blood donors was mainly monomeric (median, 83%; range, 17 to 94%). In both the patients and the positive controls, IgA1 was the predominant (greater than 85%) subclass involved, even when the IgA antibody response was mainly polymeric. Our results suggest that polymeric IgA antibody responses are linked to a strong or persisting antigenic stimulation or both. Polymeric IgA antibodies appear to be a potential marker of acute C. jejuni infections, and their determination could provide a useful tool for the serological diagnosis of recent C. jejuni infections.
Serodiagnosis of rabies by dot immunobinding assay
Authors: Heberling R L , Kalter S S , Smith J S , Hildebrand D G
Published: January 2020

A dot immunobinding assay that uses inactivated antigen for the detection of rabies viral antibodies was compared with the rapid fluorescent focus inhibition test. Results of testing pre- and postvaccination sera from humans (n = 33) and canines (n = 22) were identical for both tests. Endpoint titers of positive sera also were approximately the same by both methods. When a mouse monoclonal antibody was used, the dot immunobinding assay antigen was shown to possess detectable rabies virus glycoprotein and core antigens.
Double-antibody sandwich enzyme-linked immunosorbent assay for rapid detection of toxin-producing Corynebacterium diphtheriae
Authors: Nielsen P B , Koch C , Friis H , Heron I , Prag J , Schmidt J
Published: January 2020

An enzyme-linked immunosorbent assay for determining the toxigenicity of Corynebacterium diphtheriae is presented. The assay uses hyperimmune horse diphtheria antitoxin as a capture antibody and mouse monoclonal diphtheria antitoxin as a detecting antibody. Growth of bacteria and capture of diphtheria toxin by antitoxin are carried out in one step. Toxin produced by as little as 100 toxin-producing corynebacteria is detectable, corresponding to a sensitivity of 10 ng of diphtheria toxin per ml. Demonstration of toxin after incubation of the bacteria for 4.75 h, as well as after 18 h, was in accordance with the modified Elek gel diffusion method and the guinea pig inoculation test. However, heavy inocula incubated overnight produced significantly lower optical density than did diluted inocula; thus, the higher optical density was used as an indicator of toxin production. A decrease in optical density was also seen by shortening the incubation time. For laboratory safety, ethanol was added to the microtiter plate wells before washing out of the bacteria. This resulted in a further decrease in optical density. Using 4.75-h incubation time gave a single false-negative result. No false-positive results were ever seen. Incubation for 18 h is suitable for large-scale screening, and 4.75 h of incubation is suitable for rapid identification of toxin-producing C. diphtheriae.
Characteristics of Yersinia enterocolitica isolated from children with diarrhea in Italy
Authors: Mingrone M G , Fantasia M , Figura N , Guglielmetti P
Published: January 2020

Of 2,500 fecal samples collected from children with diarrhea in the province of Siena, 35 (1.4%) were found to be positive for Yersinia enterocolitica. Of the isolates, 94.2% belonged to biotype 4, serotype 0:3; 2.8% belonged to biotype 2, serotype O:9; and 2.8% belonged to biotype 1, serotype O:6. The in vitro pathogenicity tests showed that all but two isolates were calcium dependent and autoagglutinable and that all but one were also invasive in HEp-2 cell culture. As regards plasmid content, 32 of 33 biotype 4, serotype O:3 strains harbored a plasmid of 48 megadaltons and 1 strain also harbored a small plasmid of 2 megadaltons. The biotype 2, serotype O:9 strain harbored a plasmid of 42 megadaltons; one of the two strans lacking plasmids belonged to biotype 1, serotype O:6, and the other belonged to biotype 4, serotype O:3. Pyrazinamidase activity was positive only for the biotype 1, serotype O:6 strain. Esculin was hydrolyzed only by the biotype 1, serotype O:6 strain.
Plaque staining assay for non- or weakly cytotoxic human immunodeficiency virus
Authors: Matsui T , Nakashima H , Yoshiyama H , Kobayashi N , Yamamoto N
Published: January 2020

Cells infected with human immunodeficiency virus (HIV) were selectively stained with peroxidase-coupled antibodies in a recently developed plaque assay for HIV. The numbers of plaques formed with the human T-cell lymphotropic virus type III strain of HIV were exactly the same in stained (immunologically detectable) and unstained (visible) dishes. However, four times more plaques were visualized in stained dishes than in unstained dishes when the YU-6 and acquired immune deficiency syndrome-associated retrovirus strains of HIV were used. Linear relationship was observed between the number of stained plaques and the virus concentrations in the titration of human T-cell lymphotropic virus type III and YU-6. The assay should be useful for the titration of HIV, especially for non- or weakly cytopathic strains of HIV.
Delivery volumes of the 1- and 3-mm pins of a Cathra replicator
Authors: Rousseau D , Harbec P S
Published: January 2020

This study was undertaken to determine the volumes delivered by the 1- and 3-mm pins of a Cathra replicator and to evaluate reproducibility. The weight of broth transferred by these pins served to assess the delivery volume. Repeated transfers with the 1- and 3-mm pins showed, on average, a delivery of 0.1 and 2 microliters, respectively.
Comparison between indirect immunofluorescence and microneutralization for detection of antibodies to polioviruses
Authors: Pettit C , Minnich L L , Shehab Z M , Ray C G
Published: January 2020

Indirect immunofluorescence and microneutralization methods for the detection of antibodies to poliovirus serotypes 1, 2, and 3 were compared. Of the 41 sera tested for poliovirus type 1 antibody, 40% were in complete agreement, 55% differed by one dilution, and 5% differed by two dilutions. For poliovirus type 2, 37 sera were tested; 56% completely agreed, and 44% differed by only one dilution. For poliovirus type 3, complete agreement occurred in 59% of 33 sera, while 41% differed by one dilution. No false-negative results were obtained. These findings suggest that indirect immunofluorescence for poliovirus types 1, 2, and 3 is as sensitive as the microneutralization method and could represent a less cumbersome alternative.
Kinetics study of immunological response to Clostridium botulinum toxin
Authors: Dezfulian M , Bitar R A , Bartlett J G
Published: January 2020

A double-antibody enzyme-linked immunosorbent assay (ELISA) for detection of humoral antibody to type A botulinal toxin was developed. This assay was used to study the kinetics of antibody response of a volunteer to botulinal toxoid. The circulating type A antitoxin was first detected by the ELISA 2 weeks after the first booster injection of the toxoid. The antibody titer stayed level until the second booster at 12 weeks. The titer then continued to rise throughout the remaining study period. The neutralizing antibody to type A toxin was detected by mouse assay 15 weeks after detection of antitoxin by the ELISA.
Human immunoglobulin G subclass immune response to outer membrane antigens in meningococcal group B vaccine
Authors: Wedege E , Michaelsen T E
Published: January 2020

The immunoglobulin G (IgG) subclass distribution of antibodies against the major outer membrane proteins from serotype 2a Neisseria meningitidis in human vaccinees was studied by immunoblotting. The volunteers received two doses of a noncovalent complex of group B polysaccharide and outer membrane material from the same meningococcal strain. Six weeks after the first vaccination the antibodies mounted against the class 1 and 5 proteins belonged mainly to the IgG1 and IgG3 subclasses. However, the binding of IgG3 to the class 5 proteins decreased markedly in serum samples taken after 25 weeks. Antibody binding to the serotype-specific class 2 protein was dependent on renaturation of the antigen by a dipolar ionic detergent (R. E. Mandrell and W. D. Zollinger, J. Immunol. Methods 67:1-11, 1984). The immune response against this protein showed more individual variation and consisted of IgG1 or IgG3 or both, often combined with IgG4.
Comparison of methods for detection of serum antibody to murine rotavirus
Authors: Ferner W T , Miskuff R L , Yolken R H , Vonderfecht S L
Published: January 2020

Mice are frequently used as animal models for the study of rotaviral infections. Since natural infection is common in laboratory mice, it is important that rotaviral studies, as well as other studies utilizing suckling mice, employ animals of known immune status to murine rotavirus. A variety of homologous and heterologous enzyme immunoassay systems and an immunofluorescence technique were thus compared to determine the immunoassay that is most effective at detecting adult mice seropositive for rotaviral antibody. It was determined that a homologous enzyme immunoassay inhibition technique utilizing murine rotavirus-derived reagents was the most efficient serologic assay evaluated. A serologic response was consistently detected by this assay by 5 days after experimental rotaviral inoculation of adult mice. A homologous antibody-binding enzyme immunoassay, a heterologous inhibition enzyme immunoassay utilizing antigenically related simian rotavirus (SA-11) reagents, and an immunofluorescence technique utilizing Nebraska calf diarrhea virus antigens were found to be less sensitive for detecting serum antibody to murine rotavirus.
Nonproduction of toxic shock syndrome toxin 1 by coagulase-negative staphylococci
Authors: Parsonnet J , Harrison A E , Spencer S E , Reading A , Parsonnet K C , Kass E H
Published: January 2020

We tested 187 strains of coagulase-negative staphylococci (CNS) for the production of toxic shock syndrome toxin 1 (TSST-1). A total of 111 CNS strains were isolated from the tampons of menstruating women and 74 were isolated from unused tampons. Two strains were isolated from the genital tract of a patient with toxic shock syndrome. Strains were cultivated by the membrane-over-agar method to enhance production of TSST-1, and culture supernatants were tested by two exquisitely sensitive enzyme-linked immunosorbent assays. None of the 187 CNS strains produced TSST-1. We conclude that CNS colonizing the genital tracts of menstruating women and unused tampons produce TSST-1 infrequently, if ever, and are unlikely to play a role in toxic shock syndrome.
Subclass restriction of human enterovirus antibodies
Authors: Torfason E G , Reimer C B , Keyserling H L
Published: January 2020

We studied antibodies to enteroviruses in four groups of serum specimens: those from healthy adults, cord blood specimens, serum specimens known to contain immunoglobulin M (IgM) to coxsackie B (CB) viruses by radioimmunoassay, and serum specimens from children with symptomatic enteroviral infections. Enzyme-linked immunosorbent assays (ELISAs) were developed to detect the IgG class- and subclass (IgG1, IgG2, IgG3, and IgG4)-specific responses to CB3. The CB3 virus ELISA was not type specific. There was very poor correlation between CB3 virus neutralizing titer and IgG anti-CB3 virus ELISA results, indicating that antibodies to heterologous picornaviruses cross-react with CB3 virus in the assay. All serum specimens tested except one were IgG positive for CB3 virus. All 32 cord serum specimens were positive for IgG1 and IgG3. No enterovirus-specific IgG2 or IgG4 was detected in any serum specimen tested. Most serum specimens from the IgM-positive group, healthy adults, and children with enterovirus infections were positive for IgG1 and IgG3. Class and subclass antibody titers remained constant over time. IgG antibodies to enteroviruses appear to be restricted to the IgG1 and IgG3 subclasses. This pattern is similar to results obtained by other investigators evaluating IgG subclass antibodies to protein antigens.
Analysis of 506 consecutive positive serologic tests for brucellosis in Saudi Arabia
Authors: Kiel F W , Khan M Y
Published: January 2020

Brucellosis is common in the central region of Saudi Arabia. The major clue to the diagnosis is serologic testing. In 3 years, over 500 positive patients who had a titer of 1:160 or more were identified. Brucella species was the single organism most commonly recovered in all blood cultures, being found in 24% of patients with cultures positive for pathogenic bacteria. The expected greater number of Brucella cultures with high initial recorded titer was confirmed; 31% of the positive high-titer group had positive Brucella cultures. Conclusive correlation was uncommon below the 1:640 titer. The lower titers were less useful as indicators of active disease in this population, in which many people appear to have persistent low antibody levels.
Adherence of Pseudomonas aeruginosa to hydrophilic contact lenses and other substrata
Authors: Miller M J , Ahearn D G
Published: January 2020

Nonmucoid Pseudomonas aeruginosa isolated from corneal ulcers and contact lens cases and solutions were examined for their ability to adhere to polystyrene, glass, and hydrophilic contact lenses of varying water content and polymer composition. Adherence to the various substrates was strain specific. Adherence also was influenced by hydrophobicity and chemical composition of the substratum, as well as pH and electrolyte concentration. An extracellular polymeric adhesive appeared to be involved in the firm attachment of cells to soft contact lenses.
Improvement of serological diagnosis of neonatal cytomegalovirus infection by simultaneously testing for specific immunoglobulins E and M by antibody-capture enzyme-linked immunosorbent assay
Authors: Nielsen S L , Rønholm E , Sørensen I , Jaeger P , Andersen H K
Published: January 2020

This study describes the results of testing 92 serum samples, including 10 umbilical cord serum samples, from 38 cytomegalovirus (CMV)-infected neonates for CMV-specific immunoglobulins E (IgE) and M by antibody-capture enzyme-linked immunosorbent assays with enzyme-labeled CMV antigen. All infants excreted CMV in the urine. It was demonstrated that the CMV IgE test was more sensitive than the CMV IgM test in diagnosing CMV infection in neonates by serology. Thus, the sensitivity for the IgE test was 82%, whereas for the IgM test it was only 66%. Furthermore, the CMV-specific IgE response, expressed as absorbance, was higher than the specific IgM response in 91% of the 76 sera which contained antibodies of either one or both immunoglobulin classes. Forty-six control sera, including 18 umbilical cord sera, from 46 neonates from whom CMV was not isolated were also tested. Most sera were negative. Three infants, however, had CMV antibodies of one or both classes, indicating infection. The level of total serum IgE was controlled in 24 of the sera from the CMV-infected neonates, but in none of the cases was the level elevated. No correlation was found between the reactivity of the antibodies in the two immunoglobulin classes and the level of CMV in urine.
Unique p24 epitope marker to identify multiple human immunodeficiency virus variants in blood from the same individuals
Authors: Bolton V , Pedersen N C , Higgins J , Jennings M , Carlson J
Published: January 2020

The human immunodeficiency virus (HIV) was isolated from the blood of 192 of 410 seropositive individuals. Original isolations were made in peripheral blood mononuclear cell (PBMC) cultures, and only one-fifth of the HIV isolates could be adapted to replicate in continuous T-cell lines. Of the 192 HIV isolates, 42 had the characteristic p24 antigen marker of the acquired immunodeficiency syndrome-associated retrovirus type 2 strain of HIV (HIV ARV-2) and 150 resembled the human T-cell lymphotropic virus type III strain of HIV (HIV HTLV-III). Significantly, primary PBMC cultures from two patients yielded multiple variants. When these variants were exposed to continuous T-cell lines, only one of them continued to replicate. The remaining variants were lost and could not be reisolated following passage back into PBMC cultures. We conclude the following from these studies: PBMC cultures are more efficient at isolating HIV than continuous T-cell lines are; some patients harbor more than one genetic variant of HIV in the blood at the same time; and continuous T-cell lines are likely to yield only a portion of the HIV variants originally present in the blood.
Recovery of Blastomyces dermatitidis from blood of a patient with disseminated blastomycosis
Authors: Musial C E , Wilson W R , Sinkeldam I R , Roberts G D
Published: January 2020

This report describes the first well-documented reported case of the recovery of Blastomyces dermatitidis from the blood of a patient with disseminated blastomycosis. The isolate was recovered by using a lysis-centrifugation method and inhibitory mold agar after 6 days of incubation at 30 degrees C. One of six blood samples collected yielded a positive culture.
Evaluation of passive particle agglutination test for antibody to human immunodeficiency virus
Authors: Yoshida T , Matsui T , Kobayashi S , Yamamoto N
Published: January 2020

A gelatin particle agglutination assay was compared with indirect immunofluorescence by using 663 serum samples from acquired immunodeficiency syndrome and acquired immunodeficiency syndrome-related complex patients and asymptomatic male homosexuals in the United States and from hemophiliacs and healthy adult controls in Japan. The results showed that all 104 samples which were positive by indirect immunofluorescence were also positive by particle agglutination, while 5 additional samples were positive by particle agglutination only. The coincidence rate for antibody-positive and antibody-negative specimens was 99% (658 of 663) between particle agglutination and immunofluorescence. Four of the five samples which were positive by particle agglutination only were found by radioimmunoprecipitation to contain anti-env gene products of human immunodeficiency virus. Antibody titers of samples giving a positive reaction by particular agglutination varied from low (titer, 256) to remarkably high (256 X 10(5)). All specimens having particle agglutination titers of more than 10(5) were positive by immunofluorescence. A high correlation (r = 0.66) was observed between the titers of antibodies determined by particle agglutination and those determined by immunofluorescence. After fractionation of a serum sample from an individual at high risk by using high-performance liquid chromatography, it was shown that immunoglobulin M as well as immunoglobulin G human immunodeficiency virus antibody was detected by particle agglutination. Additional serum samples with a potential risk of giving false-positive results, such as heat-treated specimens, specimens containing antibodies to HLA, specimens containing auto-antibodies, and serum samples from individuals with a history of multiple blood transfusions, were shown to be clearly negative by particle agglutination.