bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative antigenic studies of species of Basidiobolus and other medically important fungi
Authors: Yangco B G , Nettlow A , Okafor J I , Park J , Te Strake D
Published: January 2020

An immunodiffusion technique was used to evaluate the antigenic relationship of various pathogenic and saprobic Basidiobolus spp., Conidiobolus spp., isolates of the order Mucorales, and several other medically important nonzygomycetous fungi. The antiserum to Basidiobolus haptosporus shared two lines of identity, designated inner (N) and outer (Y), when tested against exoantigens of known strains of B. haptosporus and Basidiobolus ranarum as well as exoantigens of a human Nigerian isolate and several wild isolates tentatively identified in B. ranarum. Both bands were heat stable at 56 degrees C for 30 min. Exoantigens of strains of Basidiobolus meristosporus, Basidiobolus microsporus, Conidiobolus incongruus, Conidiobolus coronatus, and other wild isolates of Basidiobolus spp. tested formed only the N immunoprecipitin band. Exoantigens of 10 isolates from other taxa did not produce any cross-reactive precipitin line. B. meristosporus antiserum that was tested against exoantigens of Basidiobolus spp. and of Conidiobolus spp. developed only an N band without a Y band. These data suggest that B. haptosporus and B. ranarum are antigenically similar to each other and distinct from B. meristosporus. Basidiobolus spp. and Conidiobolus spp. share a common N immunoprecipitin band, which implies a taxonomic relationship between these two genera. The absence of lines of identity between Basidiobolus spp. and other fungi tested suggests that, antigenically, Basidiobolus is a distinct genus.
Seroepidemiology of Klebsiella bacteremic isolates and implications for vaccine development
Authors: Cryz S J , Mortimer P M , Mansfield V , Germanier R
Published: January 2020

The frequencies of capsular serotypes among 703 Klebsiella strains isolated from the blood of hospitalized patients were determined. More than 90% of the isolates were typeable, with 69 of the 77 known serotypes being identified. Serotypes 2, 21, and 55, representing 8.9, 7.8, and 4.8% of all the isolates, respectively, were observed at a frequency significantly higher (P less than 0.05) than that for other capsular serotypes. Approximately 43% of the serotypes appeared at a frequency of less than 0.5%. Differences were found when the seroepidemiology of North American and European isolates was compared. The current findings indicate that a capsular polysaccharide-based vaccine against Klebsiella organisms is feasible and should be multivalent, eliciting antibodies directed against the 25 serotypes which make up approximately 70% of all the bacteremic isolates.
In vitro and in vivo study of stone formation by Corynebacterium group D2 (Corynebacterium urealyticum)
Authors: Soriano F , Ponte C , Santamaría M , Castilla C , Fernández Roblas R
Published: January 2020

Corynebacterium group D2 inoculated into normal human urine formed struvite crystals and an increase in pH and ammonium concentration after 24 h of incubation. Zinc disks dipped into a broth culture of this microorganism and inserted into the bladders of rats produced stones with a mean weight of 12.5 mg (ranging from 1 to 57.7 mg) after 12 days. Analysis of the infrared spectrum determined the stones to be composed of struvite. From these results its seems that stone formation by Corynebacterium group D2 may be possible both in vitro and in vivo, which may confirm a previous report involving these bacteria in human clinical encrusted cystitis.
Performance characteristics of a commercially prepared biphasic blood culture bottle
Authors: Weckbach L S , Staneck J L
Published: January 2020

A biphasic blood culture bottle (BiPB: GIBCO Laboratories, North Andover, Mass.) with an architectural design that physically separates the agar slant from the broth was compared with a conventional vented monophasic bottle (MPB-A) for use in the routine culture of blood. Both bottles contained tryptic soy broth. Tryptic soy agar was used for the BiPB slant. A third unvented bottle (MPB-N) with Columbia broth was included as part of the blood culture set. Of 3,537 sets collected, 444 were positive; 57 of these 444 sets were positive by virtue of an exclusively positive anaerobic bottle. Both BiPB and MPB-A were positive in 235 of the remaining 387 positive sets. A total of 521 isolates was recovered during the study. Of these isolates, 252 were recovered in both the BiPB and the MPB-A from the same set; 105 isolates grew in the BIPB but not in MPB-A, 95 isolates grew only in the MPB-A but not in BiPB, and 69 grew exclusively in the MPB-N. The BiPB allowed more rapid recovery of Candida spp., J-K diphtheroids, Pseudomonas spp. Making BiPB subcultures was easy enough to permit both early and daily subculture, which provided isolated colonies sooner than could be done by using the MPB-A. Isolated colonies and, therefore, identification and susceptibility results were available at least 1 day earlier for the BiPB isolates in approximately 50% of instances when both the BiPB and the MPB-A were positive. Staphylococcus epidermidis and streptococci were recovered more frequently in the BiPB, while gram-positive anaerobes were detected at a significantly (P less than 0.025) more frequent rate in the MPB-A than in the BiPB. Either bottle, however, should be used in conjunction with an anaerobic bottle for optimal recovery of anaerobic bacteria.
Comparative evaluation of four systems for determining susceptibility of gram-positive organisms
Authors: Henry D , Kunzer L , Ngui-Yen J , Smith J
Published: January 2020

A study was undertaken to compare four commercial systems for testing the antimicrobial susceptibility patterns of gram-positive cocci. The reference method was an agar dilution method. The systems evaluated were the MS-2 system (Abbott Diagnostics Div., Mississauga, Ontario), the AutoMicrobic system (AMS) (Vitek Systems, Inc., Hazelwood, Mo.) with the gram-positive susceptibility (GPS) card, the Sceptor system (BBL Microbiology Systems, distributed by Becton Dickenson, Canada Inc., Mississauga, Ontario), and the Micro-Media system (Beckman Instruments, Inc., Anaheim, Calif.). There was a greater than 98% essential accord (EA) between all test results and the reference method results when testing 134 isolates of Staphylococcus aureus. In testing 79 isolates of coagulase-negative staphylococci the EA was greater than 97% with all systems except the MS-2. In the MS-2 system only, 30% of tests were interrupted by the instrument because of insufficient growth in the control chamber. Excluding the Sceptor system, the EA was greater than 96% on testing 70 isolates of enterococcus. In testing 15 isolates of group B Streptococcus there was 91% EA with the AMS and Sceptor systems and only 71 and 88% EA with the MS-2 and Micro-Media systems, respectively. The new AMS GPS MIC card was tested against 29 methicillin-resistant S. aureus, 10 coagulase-negative staphylococci, and 9 enterococci, and it gave more accurate results than the earlier GPS breakpoint card. The Micro-Media and MS-2 systems did not reliably detect marginally methicillin-resistant S. aureus. The MS-2 was the least expensive system to operate on a cost per test basis ($3.59 Can.), whereas the Sceptor was the most expensive system ($5.29 Can.). The AMS ws the least labor intensive (0.9 min per test), and the Sceptor system was the most time consuming (2.9 min per test).
An outbreak of group B meningococcal disease: tracing the causative strain of Neisseria meningitidis by DNA fingerprinting
Authors: Kristiansen B E , Sørensen B , Bjorvatn B , Falk E S , Fosse E , Bryn K , Frøholm L O , Gaustad P , Bøvre K
Published: January 2020

Following an outbreak of meningococcal disease in three schoolchildren in a small community in northern Norway, DNA fingerprinting, serotyping with monoclonal antibodies, serogrouping, and sulfonamide sensitivity testing were applied for characterization and tracing of the causative agent. The three case isolates were genomically indistinguishable, sulfonamide-resistant, serogroup B, serotype 15 meningococci. Throat specimens were collected from 552 healthy contacts, including all children below age 17 and their parents. Among the 36 carrier isolates (carrier rate, 6.5%) 13 showed DNA fingerprints identical, or almost identical, to the index pattern. All of these 13 isolates were sulfonamide resistant, 12 were of serotype 15, and 8 were of polysaccharide serogroup B (5 were nongroupable). These closely related isolates were almost exclusively recovered from schoolchildren of 2 of 15 small villages, one of which included the homes of two of the patients. The remaining 23 carrier isolates were nonresistant, non-type 15 meningococci of widely differing DNA restriction patterns. Our results confirm that DNA fingerprinting has potential as an efficient tool in practical meningococcal epidemiology.
Differences in antibody responses with rapid agglutination tests for the detection of rubella antibodies
Authors: Chernesky M A , DeLong D J , Mahony J B , Castriciano S
Published: January 2020

Rapid agglutination tests (Rubaquick, Rubascan, and Rubacell) were used to screen sera (374 from immune and 124 from susceptible patients) for rubella immunity. Compared with enzyme immunoassay (Rubazyme) the erythrocyte agglutination assays (Rubaquick and Rubacell) were greater than 98% sensitive and 92 to 96% specific. The latex test (Rubascan) was sensitive (97.8%) and specific (96.8%) on undiluted serum but only 68.7% sensitive on serum diluted 1:10. Although the three rapid assays detected a substantial number of positive sera within 3 months of rubella immunization, a large number of variable responses were seen after infection with rubella. Analysis of discordant results suggests that these tests may be effectively used for immunity screening. The different individual assay results observed on low-titered sera or blood collected shortly after infection or immunization may not be comparable, because each assay has a different antigenic component on the agglutinin.
Separation of cryptosporidium oocysts from fecal debris by density gradient centrifugation and glass bead columns
Authors: Heyman M B , Shigekuni L K , Ammann A J
Published: January 2020

A method was developed to obtain purified cryptosporidium oocysts from fecal samples. Oocysts were initially collected by centrifugation through a sucrose density gradient and further purified by passage through glass bead columns. The purified oocysts were antigenically active and sufficiently pure for immunological studies.
False-positive results with hepatitis B virus DNA dot-hybridization in hepatitis B surface antigen-negative specimens
Authors: Diegutis P S , Keirnan E , Burnett L , Nightingale B N , Cossart Y E
Published: January 2020

Three serum samples derived from healthy hepatitis B surface antigen-negative subjects were found to be reactive for hepatitis B virus (HBV) DNA sequences when assayed by DNA dot-hybridization. All three results were shown to be due to the presence of sequences which reacted with residual bacterial plasmid vector sequences in the DNA probe, and no evidence of HBV markers was demonstrated in the sera. This is the first report of a false-positive result with the HBV DNA dot-hybridization assay.
Analysis by coagglutination of the distribution of a 24,000-dalton surface protein in Yersinia isolates
Authors: Toyos J , Díaz R , Urra E , Moriyón I
Published: January 2020

The presence of a 24,000-dalton surface protein in 215 isolates of Yersinia enterocolitica and related species was examined. By coagglutination with the specific antiserum, a 100% correlation with the pathogenic biogroups was found. Thus, this method is useful for the rapid screening of potential pathogenic Y. enterocolitica isolates.
Staphylococcal peritonitis in patients on continuous peritoneal dialysis
Authors: West T E , Walshe J J , Krol C P , Amsterdam D
Published: January 2020

During 1984, 35 patients undergoing continuous peritoneal dialysis experienced 77 cases of peritonitis with 55 cases (71.4%) related to staphylococci. Coagulase-negative staphylococci were isolated in 41 cases, while Staphylococcus aureus was found in 14. A coexisting tunnel infection was more often associated with S. aureus (7/14) than with coagulase-negative staphylococci (2/41) (P less than 0.01). Likewise, eradication of the infection necessitated catheter removal more frequently with S. aureus (5/14) than with coagulase-negative staphylococci (2/41) (P less than 0.01). Of the 41 coagulase-negative staphylococci, 35 were characterized as to species, adherence, and production of two exopolysaccharides. Staphylococcus epidermidis was the most frequent coagulase-negative species (29/35). Peritonitis cases caused by coagulase-negative staphylococci that lacked adherence and exopolysaccharides were more frequently associated with complications (4/6) than were those organisms with either or both properties of adherence or exopolysaccharide production (5/29). There were no appreciable differences in antibiotic susceptibilities. Staphylococcal peritonitis remains a significant cause of morbidity in continuous peritoneal dialysis patients. The incidence of complications was not directly linked to staphylococcal properties of adherence or exopolysaccharide production.
Comparison of Chalquest and Hayflick media, with and without ammonium reineckate, for isolating Mycoplasma pulmonis from rats
Authors: Lai W C , Pakes S P , Stefanu C , Lu Y S
Published: January 2020

Chalquest and Hayflick media with and without ammonium reineckate were compared for isolation of Mycoplasma pulmonis from the nasopharyngeal ducts, tracheobronchial trees, and middle ears of 66 naturally infected rats. The results show that 92% (366 of 396) of the samples were positive for M. pulmonis in Chalquest medium with and without ammonium reineckate and 66% (260 of 396) were positive in Hayflick medium with and without ammonium reineckate (P less than 0.001). An enhancing effect of ammonium reineckate on M. pulmonis isolation was observed only in Hayflick medium; the isolation rate was 76% (151 of 198) in Hayflick medium with ammonium reineckate as compared with 55% (109 of 198) in Hayflick medium without ammonium reineckate (P less than 0.03). No significant differences in isolation rates were observed between Chalquest medium with and without ammonium reineckate. The mean growth time of M. pulmonis on Chalquest medium was 3.4 days as compared with 5.1 days in Hayflick medium, indicating that M. pulmonis can be detected earlier on Chalquest medium than on Hayflick medium. These data indicate that Chalquest medium is superior to Hayflick medium for M. pulmonis isolation from rats.
Opportunistic infection of the spleen caused by Aureobasidium pullulans
Authors: Salkin I F , Martinez J A , Kemna M E
Published: January 2020

The mold Aureobasidium pullulans was isolated on several nutrient media from a splenic abscess in a patient with disseminated lymphoma. Examination of stained smears and paraffin sections revealed fungal structures characteristic of this organism. This is the first reported association of A. pullulans with an opportunistic visceral infection.
Development and application of monoclonal antibodies for specific detection of human enteric adenoviruses
Authors: Singh-Naz N , Naz R K
Published: January 2020

Monoclonal antibodies were prepared against enteric adenovirus by fusing P3-NS1/-Ag4-1 mouse myeloma cells with lymphocytes from BALB/c mice immunized with enteric adenovirus 40 (Ad40) G2297. Of the several putative clones secreting antibodies to adenovirus, five were found to react specifically to the enteric adenovirus. The specificity of two of these monoclones which recognize a single antigen of a molecular size of 17 kilodaltons was evaluated against 78 clinical isolates. One monoclone (5D8/2C2) reacted with both Ad40 and Ad41, and the other monoclone (2H6/C11) recognized Ad40 only in an enzyme-linked immunosorbent assay (ELISA). These ELISA results correlated well with those of the specific neutralization test or DNA restriction endonuclease analysis or both. The use of this rapid ELISA with these monoclones will find applications in the diagnosis of enteric adenovirus and should facilitate the epidemiologic studies of enteric adenovirus gastroenteritis.
Evaluation of live avian-human reassortant influenza A H3N2 and H1N1 virus vaccines in seronegative adult volunteers
Authors: Snyder M H , Clements M L , Betts R F , Dolin R , Buckler-White A J , Tierney E L , Murphy B R
Published: January 2020

An avian-human reassortant influenza A virus deriving its genes coding for the hemagglutinin and neuraminidase from the human influenza A/Washington/897/80 (H3N2) virus and its six "internal" genes from the avian influenza A/Mallard/NY/6750/78 (H2N2) virus (i.e., a six-gene reassortant) was previously shown to be safe, infectious, nontransmissible, and immunogenic as a live virus vaccine in adult humans. Two additional six-gene avian-human reassortant influenza viruses derived from the mating of wild-type human influenza A/California/10/78 (H1N1) and A/Korea/1/82 (H3N2) viruses with the avian influenza A/Mallard/NY/78 virus were evaluated in seronegative (hemagglutination inhibition titer, less than or equal to 1:8) adult volunteers for safety, infectivity, and immunogenicity to determine whether human influenza A viruses can be reproducibly attenuated by the transfer of the six internal genes of the avian influenza A/Mallard/NY/78 virus. The 50% human infectious dose was 10(4.9) 50% tissue culture infectious doses for the H1N1 reassortant virus and 10(5.4) 50% tissue culture infectious doses for the H3N2 reassortant virus. Both reassortants were satisfactorily attenuated with only 5% (H1N1) and 2% (H3N2) of infected vaccines receiving less than 400 50% human infectious doses developing illness. Consistent with this level of attenuation, the magnitude of viral shedding after inoculation was reduced 100-fold (H1N1) to 10,000-fold (H3N2) compared with that produced by wild-type virus. The duration of virus shedding by vaccines was one-third that of controls receiving wild-type virus. At 40 to 100 50% human infectious doses, virus-specific immune responses were seen in 77 to 93% of volunteers. When vaccinees who has received 10(7.5) 50% tissue culture infectious doses of the H3N2 vaccine were experimentally challenged with a homologous wild-type human virus only 2 of 19 (11%) vaccinees became ill compared with 7 of 14 (50%) unvaccinated seronegative controls ( P < 0.025; protective efficacy, 79%). Thus, three different virulent human influenza A viruses have been satisfactorily attenuated by the acquisition of the six internal genes of the avian influenza A/Mallard/NY/78 virus. The observation that this donor virus can reproducibly attenuate human influenza A viruses indicates that avian-human influenza A reassortants should be further studied as potential live influenza A virus vaccines.
Slime production by bovine milk Staphylococcus aureus and identification of coagulase-negative staphylococcal isolates
Authors: Rather P N , Davis A P , Wilkinson B J
Published: January 2020

Staphylococcus aureus isolates from bovine milk were assessed for capsule or slime production. When pure S. aureus cultures in milk were inoculated directly into serum-soft agar constituted with a modified staphylococcus 110 medium, 100% of the isolates grew with diffuse colony morphology. Diffuse colony morphology was rapidly lost on subculture and was more rapidly lost in brain heart infusion-serum-soft agar. No evidence was seen for encapsulation in India ink preparations or by the clumping factor test. It was concluded that freshly isolated S. aureus strains produce slime, not true capsules. During examination of the 84 milk samples that grew staphylococci in addition to S. aureus (27.4%), a significant number of coagulase-negative staphylococcal species were encountered and identified by conventional tests as S. simulans (41.7%), S. xylosus (11.9%), S. epidermidis (3.6%), S. saprophyticus (3.6%), S. hyicus (2.9%), S. cohnii (1.2%), S. haemolyticus (1.2%), and S. warneri (1.2%). Five isolates (6.0%) were not identified. Attempts were also made to identify the isolates by the API Staph-Ident system, which gave an overall accuracy of 45.2%. The susceptibilities of the isolates to a variety of antibiotics were determined, and they appeared to be less resistant than human clinical isolates.
Sorbitol-MacConkey medium for detection of Escherichia coli O157:H7 associated with hemorrhagic colitis
Authors: March S B , Ratnam S
Published: January 2020

Escherichia coli serotype O157:H7 is a recently recognized human pathogen associated with hemorrhagic colitis. Unlike most E. coli strains, E. coli O157:H7 does not ferment sorbitol. Therefore, the efficacy of MacConkey agar containing sorbitol (SMAC medium) instead of lactose as a differential medium for the detection of E. coli O157:H7 in stool cultures was determined in comparison with MacConkey agar. The relative frequency of non-sorbitol-fermenting (NSF) organisms other than E. coli O157:H7 in feces was low at 10 to 20% (95% confidence limits), and NSF organisms also occurred mostly in small numbers. In a field trial involving over 1,000 diarrheal stools, E. coli O157:H7 was isolated from 18 stools, all of which were from patients with bloody diarrhea. In every instance, the growth of E. coli O157:H7 on SMAC medium was heavy and occurred in almost pure culture as colorless NSF colonies in contrast to fecal flora, which are mostly sorbitol fermenting and hence appear pink on this medium, whereas on MacConkey agar cultures, the growth of E. coli O157:H7 was indistinguishable from fecal flora. SMAC medium permitted ready recognition of E. coli O157:H7 in stool cultures. Detection of E. coli O157:H7 on SMAC medium had a sensitivity of 100%, a specificity of 85%, and an accuracy of 86%. SMAC medium stool culture is a simple, inexpensive, rapid, and reliable means of detecting E. coli O157:H7, and we recommend routine use of SMAC medium especially for culturing bloody stools.
Evaluation of the Minitek gram-positive set for identification of staphylococci isolated from the bovine mammary gland
Authors: Watts J L , Owens W E , Nickerson S C
Published: January 2020

The Minitek Gram-Positive Set was evaluated as a means of identifying staphylococci isolated from bovine mammary glands. Initial accuracy was 79.2%. Misidentification of isolates due to data base deficiencies resulted with the animal-associated species Staphylococcus intermedius and S. hyicus. Minor modification to account for data base deficiencies permitted recognition of 87.7% of the isolates. Incorporation of additional veterinary isolates into the data base would improve the accuracy of the Minitek system and enhance acceptance by veterinary microbiologists.
Treponemal antibody-absorbent enzyme immunoassay for syphilis
Authors: Chen J , Lin T M , Schubert C M , Halbert S P
Published: January 2020

An enzyme immunoassay for the diagnosis of syphilis (ELISA-SY) was developed with solid-phase extracts of Treponema pallidum, specimen diluent containing Reiter treponeme absorbent, and three 30-min incubations. The ELISA-SY results were determined in comparison with a standardized positive control and reported as a percentage of strong positive control. In tests with 1,005 serum samples from a venereal disease clinic and other sources, 98.2% agreement was found with fluorescent treponemal antibody-absorption (FTA-ABS) results, and 98.3% agreement was found with T. pallidum passive hemagglutination (PHA) findings. Only 1 of 29 sera originally considered to be biologically false-positive by ELISA-SY; the latter specimen was also positive by PHA and FTA-ABS tests performed in our laboratories. Serum samples from clinically diagnosed syphilitics (16 primary-stage isolates, 7 secondary-stage isolates, and 3-latent-stage isolates) were all positive by ELISA-SY, FTA-ABS, and PHA. Serum samples from 51 newborns suspected of having syphilis on the basis of positive cardiolipin flocculation tests showed 98% agreement of ELISA-SY results with FTA-ABS and PHA findings. Sera from all 61 patients with a variety of autoimmune and other diseases known to be associated with biologically false-positive reactions for syphilis were negative by this ELISA-SY. The specificity of the ELISA procedure for T. pallidum antibody was also confirmed immunologically by blocking experiments.
Enrichment for detection of Campylobacter jejuni
Authors: Shimada K , Tsuji H
Published: January 2020

Gifu anaerobe-modified semisolid medium supplemented with vancomycin, polymyxin B, trimethoprim, and amphotericin B can serve as an efficient enrichment for the recovery of Campylobacter jejuni under aerobic conditions. It is superior to the direct plating method in its recovery of very small numbers of C. jejuni. Blood supplementation, agitation, and elevated temperatures are not required for this improved recovery.
Characteristics of Yersinia intermedia-like bacteria isolated from patients with diarrhea in Nigeria
Authors: Agbonlahor D E
Published: January 2020

Five Yersinia strains, biochemically mimicking Yersinia intermedia, were isolated from patients with acute diarrheal diseases seen at the University of Lagos (Nigeria) Teaching Hospital between October 1979 and September 1982. The five isolates utilized citrate as their source of carbon and fermented sucrose, L-rhamnose, and alpha-methyl-D-glucoside, but unlike any other Y. intermedia biogroup, they failed to ferment D-melibiose or D-raffinose. They all belonged to serogroup O:52,53. Four autoagglutinated and were calcium dependent for growth at 37 degrees C. Three caused conjunctivitis in the guinea pig eye model, but none possessed the 42-megadalton virulence plasmid. Four of the isolates caused diarrhea in orally infected mice and rabbits but were nonlethal for these animals. These results suggest that these isolates represent yet another Yersinia species with disease-producing capabilities in humans.
Detection of rotavirus in stool specimens with monoclonal and polyclonal antibody-based assay systems
Authors: Knisley C V , Bednarz-Prashad A J , Pickering L K
Published: January 2020

Accurate diagnosis of rotavirus is important in both clinical and research situations. A total of 100 stool specimens from children with diarrhea were tested for rotavirus by electron microscopy. These specimens were then coded and tested for rotavirus by four procedures: a monoclonal antibody-based enzyme immunoassay (EIA) (Pathfinder; Kallestad Laboratories, Inc., Austin, Tex.), two polyclonal antibody-based EIAs (Rotazyme II; Abbott Laboratories, North Chicago, Ill.; and an EIA performed with reagents from the National Institutes of Health, Bethesda, Md. [NIH reagent EIA]), and a latex agglutination (LA) assay (Rotalex; Medical Technology Corp., Somerset, N.J.). The sensitivity of the monoclonal antibody EIA (95%) was superior to those of the polyclonal antibody EIAs (73% for Rotazyme II and 57% for the NIH reagent EIA) and the LA assay (61%). The specificity of the LA assay (98%) was slightly better than those of the other systems (88 to 96%). The positive and negative predictive values of the monoclonal antibody EIA (93 and 96%, respectively) were better than those of Rotazyme II (82 and 80%, respectively), the LA assay (96 and 76%, respectively), and the NIH reagent EIA (93 and 74%, respectively). The visual readings of the monoclonal antibody EIA correlated better with the spectrophotometric optical density readings than did the visual readings of the polyclonal antibody EIAs; however, the agreement of both with electron microscopy results was poor when 1+ or plus-minus readings were observed. The monoclonal antibody EIA is more sensitive and predictive than other rotavirus detection systems and second only to the LA assay in specificity in detecting rotavirus in stool specimens.
Immune-specific gamma interferon production correlates with lymphocyte blastogenesis
Authors: D'Andrea A D , Plotkin S A , Douglas S D , Polin R A
Published: January 2020

Gamma interferon (IFN-gamma) production, measured by a new commercially available radioimmunoassay, and lymphocyte blastogenesis were investigated in human peripheral lymphocyte cultures from healthy adults stimulated by crude cytomegalovirus (CMV) antigen. Mononuclear cells were obtained from 32 healthy adults (18 CMV seropositive [S+] and 14 CMV seronegative [S-]) by Ficoll-Hypaque gradients and cultured in microtiter plates containing CMV antigen. Lymphocyte blastogenesis ([3H]thymidine uptake) and IFN-gamma were determined on day 6. The mean stimulation index for S+ individuals was significantly greater than that for S- individuals (P less than 0.001). Similarly, the IFN-gamma stimulation index was greater for S+ individuals than for S- individuals (P less than 0.005). A significant increase in the concentration of IFN-gamma (10 NIH units/ml) was observed for S+ individuals at 24 h of antigen stimulation, with peak levels at 4 days. The radioimmunoassay for IFN-gamma production by antigen-stimulated lymphocytes in vitro (IMRX; Centocor Inc., Malvern, Pa.) is a rapid and sensitive measure of cell-mediated immunity.
Nosocomial infection and colonization by Klebsiella trevisanii
Authors: Freney J , Gavini F , Alexandre H , Madier S , Izard D , Leclerc H , Fleurette J
Published: January 2020

During an 18-month period we identified two cases of septicemia and 24 examples of colonization of humans by Klebsiella trevisanii. Organisms were identified using the API 20EC and API 147 assimilation galleries. Of 147 clinical isolates initially identified as K. oxytoca, 18% were found to be K. trevisanii. Tracheal aspirate was the most common source of the organism. An extensive environmental sampling survey in the rooms of 12 colonized patients revealed a possible reservoir of the organism only once (a face cloth).
T-cell detection with monoclonal antibody T101 kits
Authors: Pollack S M , Cimino E F , Robbins D S , Hoffman P M
Published: January 2020

A solid-phase immunoadsorption procedure (Quantigen T&B cell kit; Bio-Rad Laboratories, Richmond, Calif.) employing monoclonal antibody T101 detected mean percentages of peripheral blood T cells comparable to those obtained by rosetting with sheep erythrocytes, while lower values were obtained with an indirect immunofluorescence procedure (Cytotag T&B cell kit; Hybritech, Inc., San Diego, Calif.) employing the same antibody. Therefore, T101 binding appears to be more easily detected by solid-phase immunoadsorption than by immunofluorescence microscopy.
Pseudomonas cepacia isolated from crystal violet solution in a hospital laboratory
Authors: Walsh D M , Eberiel D T
Published: January 2020

Pseudomonas cepacia was isolated from crystal violet solution in a hospital laboratory. A similar strain was isolated from the cerebrospinal fluid of a patient. It was difficult to determine whether these strains were identical, but they possessed similar biochemical patterns.
Variations in bacitracin susceptibility observed in Staphylococcus and Micrococcus species
Authors: Baker J S , Hackett M F , Simard D J
Published: January 2020

Bacitracin susceptibility was evaluated as a laboratory method to differentiate staphylococci from micrococci. A total of 317 staphylococcal isolates and 108 micrococcal isolates were each tested for susceptibility to bacitracin by a disk-diffusion method using disks of three different potencies (0.04, 2.0, and 10.0 U) and a broth dilution method to obtain MICs. When a growth inhibition zone diameter breakpoint of greater than 10 mm was used to establish susceptibility with a 0.04-U disk, all micrococci were bacitracin susceptible and 94.6% of the staphylococci were resistant. Testing with disks of higher potency did not improve the specificity of the disk-diffusion method.
Maxillary sinus infection caused by the homobasidiomycetous fungus Schizophyllum commune
Authors: Kern M E , Uecker F A
Published: January 2020

We investigated a case of maxillary sinus infection caused by the medusoid mutant of the basidiomycetous fungus Schizophyllum commune. Morphologies of both typical and medusoid forms of S. commune were determined. Prior reports of pathogenic homobasidiomycetes are discussed.
Serum and nasal-wash immunoglobulin G and A antibody response of infants and children to respiratory syncytial virus F and G glycoproteins following primary infection
Authors: Murphy B R , Graham B S , Prince G A , Walsh E E , Chanock R M , Karzon D T , Wright P F
Published: January 2020

An enzyme-linked immunosorbent assay (ELISA) with immunoaffinity-purified fusion (F) or attachment (G) glycoprotein was used to measure the serum and secretory immune responses of 18 infants and children, 4 to 21 months of age, who underwent primary infection with respiratory syncytial virus (RSV). Most of the 10 older individuals (9 to 21 months of age) developed moderate levels of serum and nasal-wash immunoglobin A (IgA) and IgG F and G antibodies. These individuals developed a moderate level of serum or nasal-wash antibodies that neutralized virus infectivity. One of the eight younger individuals (4 to 8 months of age) failed to develop an F antibody response, while three failed to develop a G antibody response. The most notable difference in the responses of the two age groups involved the titer in convalescent sera of G, F, and neutralizing antibodies which were 8- to 10-fold lower in younger individuals. Most of the younger infants failed to develop a rise in serum or nasal-wash neutralizing antibody. It is possible that the presence of maternally derived antibody in the younger infants suppressed the immune response to RSV infection, and that this accounted, in part, for the low level of postinfection antibody titer in this group. This low level and the irregular response of the infants less than 8 months of age may contribute to the severity of their initial infection and may also be responsible, in part, for their failure to develop effective resistance to subsequent reinfection by RSV.
Appearance of specific antibody-bearing cells in human bronchial mucosa after local immunization with bacterial vaccine
Authors: Latil F , Vervloet D , Casanova P , Garbe L , Fuentes P , Wierzbicki N , Charpin J
Published: January 2020

The immune response to local in vivo inhalation of a lysed bacteria vaccine was assessed in surgical specimens of main-stem bronchi from patients who had undergone pneumectomy for cancer. The patient population included 22 subjects; 11 of these received the aerosol vaccine twice a day for 10 days prior to surgery, while the remaining 11 patients were used as controls and were not immunized. The submucous glands of immunized subjects showed significantly more cells than did those of the controls, i.e., 62 +/- 8 versus 37 +/- 7, respectively (P less than 0.05). The following five antigens were chosen for study by fluorescence assay: Streptococcus pneumoniae types II and III, Haemophilus influenzae, Streptococcus sp. strain D19, and Klebsiella pneumoniae. An immunization-dependent correlation was found between immunoglobulin A, immunoglobulin A-bearing cells, and specific antibody-bearing cells on the one hand and three of the five antigens (S. pneumoniae types II and III and Streptococcus sp. strain D19) on the other hand. This is the first time that a relationship has been established between bacterial immunization of the lower respiratory tract and local immunoglobulin production in humans.
Detection of Pseudomonas mesophilica as a source of nosocomial infections in a bone marrow transplant unit
Authors: Gilchrist M J , Kraft J A , Hammond J G , Connelly B L , Myers M G
Published: January 2020

Pseudomonas mesophilica was isolated from fungal blood cultures of two bone marrow transplant recipients who consecutively occupied the same room. The isolation of P. mesophilica was temporally associated with febrile illness in these two granulocytopenic patients at 1 and 3 weeks posttransplant. A third patient, housed separately on the same bone marrow transplant unit, had nasopharyngeal colonization by this organism. Epidemiologic risk factors in common included staff, medications, and oral and perineal irrigations with tap water. Surveillance cultures detected P. mesophilica in none of 24 pharmaceutical preparations and in 10 of 40 tap water samples (100 to 600 CFU/ml) from implicated and control rooms on the same floor. Antimicrobial susceptibility testing of 14 patients and environmental isolates by agar dilution revealed similar profiles; some environmental isolates exhibited higher MICs. Because of restrictive nutritional and temperature requirements, P. mesophilica is undetected by many clinical laboratory protocols and may represent a previously undetected source of febrile illness in neutropenic patients.
Detection of human T-cell lymphotropic virus type III-related antigens and anti-human T-cell lymphotropic virus type III antibodies by anticomplementary immunofluorescence
Authors: Blumberg R S , Sandstrom E G , Paradis T J , Neumeyer D N , Sarngadharan M G , Hartshorn K L , Byington R E , Hirsch M S , Schooley R T
Published: January 2020

Techniques presently available for detection of human T-cell lymphotropic virus type III (HTLV-III) antigens and antibodies are laborious or relatively nonsensitive. We adapted anticomplementary immunofluorescence (ACIF) for these purposes. In HTLV-III-infected cells, specific ACIF was demonstrated by a diffuse speckling pattern that often resulted in a peripheral cellular rim of fluorescence. A 97% concordance was demonstrated between the ACIF assay and other sensitive tests for HTLV-III antibody detection (Western blot and membrane immunofluorescence and fixed-cell immunofluorescence tests). The ACIF assay was both more sensitive and more specific when compared with the enzyme-linked immunosorbent assay. For detection of HTLV-III antigens, the ACIF assay appeared to be as sensitive as the reverse transcriptase assay and more sensitive, with less background reactivity, than the conventional immunofluorescence assay. The ACIF assay often detected low levels of HTLV-III antigens within 3 days of infection in vitro, compared with 5 to 7 days with the indirect immunofluorescence assay, and generally paralleled the reverse transcriptase assay. The ACIF assay is a simple, sensitive, and specific assay for detection of HTLV-III-related antigens and antibodies. It should prove useful in the diagnosis of HTLV-III infection, as well as in studies of pathogenesis.
In vitro evaluation of combined usage of fosfomycin and 5-fluorouracil for selective isolation of Leptospira species
Authors: Oie S , Koshiro A , Konishi H , Yoshii Z
Published: January 2020

The combined usage of fosfomycin (FOM) and 5-fluorouracil (5-FU) as selective agents for the isolation of leptospires from contaminated materials was investigated. Additive or synergistic antibacterial activity was apparent with the combination compared with each agent used separately. Of 54 bacterial strains tested, 52 were inhibited, while all 5 Leptospira strains tested were unaffected by the combined addition of FOM (400 micrograms/ml) and 5-FU (100 micrograms/ml) to Korthof medium. Furthermore, this combination successfully supported the selective growth of Leptospira interrogans serovar copenhageni in experimentally contaminated specimens. This FOM-5-FU combination is surmised to be useful for the selective isolation of leptospires from contaminated clinical, pathological, or environmental materials.
Synthetic oligodeoxyribonucleotide probes to detect Kanagawa phenomenon-positive Vibrio parahaemolyticus
Authors: Nishibuchi M , Hill W E , Zon G , Payne W L , Kaper J B
Published: January 2020

Synthetic oligodeoxyribonucleotide probes were used in the colony hybridization test to examine the association between the Kanagawa phenomenon (KP) and the thermostable direct hemolysin gene (tdh) of Vibrio parahaemolyticus. Representative V. parahaemolyticus strains with a variety of KP reactions and 17 other Vibrio species were examined for homology with four synthetic oligodeoxyribonucleotide probes (19 to 21 bases long) representing different regions of the tdh structural gene. Under stringent conditions, two of the probes were capable of distinguishing KP-positive V. parahaemolyticus from KP-negative or KP weak-positive V. parahaemolyticus which possesses mutated tdh genes. Vibrio hollisae strains hybridized with all four probes under reduced stringency, suggesting that they have tdh-related genes which are homologous but not identical to the tdh gene in all the regions examined. The results suggest that the colony hybridization test with the synthetic oligonucleotide probes is more suitable for the definitive determination of KP-positive strains than the hybridization with the larger gene probe or immunological assays.
In vitro study of bacterial growth in continuous ambulatory peritoneal dialysis fluids
Authors: Sheth N K , Bartell C A , Roth D A
Published: January 2020

We examined the in vitro survival of bacteria in continuous ambulatory peritoneal dialysis effluents of patients with clinical peritonitis and those without peritonitis. Standard strains of coagulase-negative staphylococci (CNS), Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were inoculated into the fluids, and portions were plated for bacterial counts at 0.5, 4, 24, 48, 72, and 96 h. Unused dialysate fluid was also inoculated simultaneously. Our results show that CNS increased minimally up to 48 h in the noninfected continuous ambulatory peritoneal dialysis effluents and decreased by 96 h, whereas survival was only minimal in the infected effluent. S. aureus showed trends similar to those of CNS, but differences in survival in infected and noninfected effluents were less marked. By contrast, E. coli and P. aeruginosa increased by greater than 1,000-fold in all solutions tested. Based on the above findings, it is likely that a proportionate number of culture-negative cases of peritonitis are due to gram-positive cocci, especially CNS, which are not retrievable by standard culture techniques because of poor survival rate.
Comparison of agar dilution, microdilution, and disk elution methods for measuring the synergy of cefotaxime and its metabolite against anaerobes
Authors: Smith J A , Henry D , Ngui-Yen J , Castell A , Coderre S
Published: January 2020

The activities of cefotaxime (CTX) and desacetyl cefotaxime (des-CTX) were tested both singly and in combination against 173 anaerobic clinical isolates. The MIC of CTX for 50% of 60 Bacteroides fragilis isolates was 22.4 micrograms/ml in broth, compared with 47.4 micrograms/ml in agar. This reduced efficacy in agar was seen with all species tested and is in apparent conflict with reported clinical efficacy of the drug. Synergy between CTX and des-CTX was observed with 70 to 100% of the isolates, including 60% of all Bacteroides spp. tested. The susceptibility results in a synergy system correlated well with those noted in a broth-disk elution method incorporating 32 micrograms of CTX and 8 micrograms of des-CTX per ml. The correlation was poorer when the broth-disk method contained 16 micrograms of CTX and 8 micrograms of des-CTX per ml.
Cryptosporidium spp., a frequent cause of diarrhea in Liberian children
Authors: Højlyng N , Mølbak K , Jepsen S
Published: January 2020

This report presents results from a sample survey designed to investigate the possible role of Cryptosporidium spp. in childhood diarrhea in a developing country, Liberia, West Africa. During the four months of January to April 1983, a house-to-house study was carried out in two geographically and socially different communities--an urban slum and three rural villages. Stool samples from 374 children, aged 6 to 59 months, were tested for Cryptosporidium spp. Among the children with diarrhea 8.4% were Cryptosporidium spp. positive compared with a prevalence rate of 5.9% in asymptomatic children. Of the children living in a household with a Cryptosporidium spp.-positive index child, 8.6% had a positive stool sample. Of all children attending a clinic because of diarrhea, 14.6% were Cryptosporidium spp. positive. Cryptosporidiosis was more frequent in younger children; 24 of the total of 29 positive cases (83%) were below 2.5 years old. Actual or previous bottle feeding (formula) was a risk factor, particularly in children below 18 months old. Of the bottle-fed children, 28% were Cryptosporidium spp. positive versus 9.1% of children never bottle fed. Crowding is another possible risk factor. The prevalence of cryptosporidiosis was 13.5% in big urban households with more than 10 children, whereas the prevalence in the small urban households was 6.1%. Ethnic and religious differences were particularly evident in the rural area. No Muslim households had cryptosporidiosis, whereas the prevalence in non-Muslim tribes was 9%. The general belief that cryptosporidiosis is primarily a zoonosis is questioned in this study, partly because many carriers and asymptomatic household contacts were found.
Penicillin resistance in the subgingival microbiota associated with adult periodontitis
Authors: Kinder S A , Holt S C , Korman K S
Published: January 2020

In this investigation, the penicillin-resistant and beta-lactamase-producing subgingival microbiota associated with adult periodontitis was identified, and the impact of a recent exposure to penicillin on the recovery of resistant organisms from this microbiota was assessed. Subjects with adult periodontitis were examined clinically and microbiologically. Twenty-one subjects had a documented history of penicillin therapy within the previous 6 months whereas an additional 21 subjects had no history of antibiotic use within 1 year. Subgingival plaque samples were cultured anaerobically on nonselective and penicillin-containing elective media. MICs and beta-lactamase production were determined for the isolates from the elective medium. The penicillin-resistant microbiota consisted primarily of gram-negative organisms, including Bacteroides, Veillonella, Haemophilus, Eikenella, and Capnocytophaga species. The prevalence (P less than 0.05) and proportions (P less than 0.005) of both penicillin-resistant pigmented Bacteroides and Veillonella species were significantly greater in subjects with recent penicillin exposure. Of the penicillin-resistant genera identified, beta-lactamase production was detected in species of pigmented Bacteroides, Capnocytophaga, and Streptococcus. The prevalence of beta-lactamase-producing Bacteroides species was significantly greater in subjects with recent penicillin exposure (P less than 0.05). Of the antibiotics examined, no single agent was uniformly effective against all of the penicillin-resistant strains, but metronidazole and clindamycin were active against all of the penicillin-resistant pigmented Bacteroides strains.
Evaluation of a monoclonal antibody test to detect chlamydia in cervical and urethral specimens
Authors: Forbes B A , Bartholoma N , McMillan J , Roefaro M , Weiner L , Welych L
Published: January 2020

The MicroTrak Chlamydia trachomatis Direct Specimen Test (MT; Syva Co., Palo Alto, Calif.) was compared with cell culture in two patient populations. The sensitivity of the MT for a low-prevalence group was significantly lower (59.6%) than that for a high-prevalence group (84.4%). The results underscored the need to run the MT in parallel with culture initially if the prevalence of chlamydial infections is unknown and questioned the usefulness of the MT as a screening test for chlamydia in low-prevalence populations.
Separation of hemagglutination-inhibiting immunoglobulin M antibody to rubella virus in human serum by high-performance liquid chromatography
Authors: Kobayashi N , Suzuki M , Nakagawa T , Matumoto M
Published: January 2020

High-performance liquid chromatography was successfully used to separate hemagglutination-inhibiting immunoglobulin M (IgM) rubella virus antibody from IgG rubella virus antibody in human serum. The fractionation by high-performance liquid chromatography was as effective as sucrose density gradient centrifugation in separating IgM antibody from IgG antibody.