bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of the biotin-avidin system to study the specificity of antibodies against respiratory syncytial virus
Authors: Anderson L J , Coombs R A , Tsou C , Hierholzer J C
Published: January 2020

To characterize the specificity of monoclonal antibodies against respiratory syncytial virus, we developed a technique which combined the biotin-avidin system, immunoprecipitation, and transblot electrophoresis. The viral proteins were first biotinylated and then immunoprecipitated with a monoclonal antibody or respiratory syncytial virus immune serum, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and electrophoretically transferred to nitrocellulose paper. The protein bands were located on the paper with avidin-peroxidase plus a precipitable substrate. This procedure gave reproducible results qualitatively similar to those obtained by radioimmunoprecipitation but without the cost, risk of radiation exposure, or disposal problems of radioisotopes. This procedure should have widespread applications.
Demonstration of uronic acid capsular material in the cerebrospinal fluid of a patient with meningitis caused by mucoid Pseudomonas aeruginosa
Authors: Stevens D , Lieberman M , McNitt T , Price J
Published: January 2020

A 39-year-old female with insulin-dependent diabetes mellitus developed Rhizopus infection of the maxillary sinus. Subsequent to successful treatment with amphotericin B and surgical debridement, she developed purulent meningitis due to a mucoid strain of Pseudomonas aeruginosa. Analysis of cerebrospinal fluid documented the presence of a uronic acid polymer at a concentration of 40 micrograms/ml. In spite of parenteral and intrathecal antibiotic therapy, the patient died. This case illustrates that mucoid strains of P. aeruginosa may result in fatal infection and that alginate capsule is produced in vivo in humans.
Bim, a new serovar of Leptospira interrogans isolated from a dog in Barbados
Authors: Jones C J , Sulzer K R , Everard C O , Vaughn A W , Innis V A
Published: January 2020

A new serovar of Leptospira interrogans was isolated from a dog in Barbados. The proposed name of the new serovar is bim, and the designated type strain is 1051. The serogroup of the new serovar is the Autumnalis serogroup. The new serovar was subsequently isolated from six patients with leptospirosis in Barbados.
Cost-effective method of triple-site culturing for Neisseria gonorrhoeae
Authors: Gartner T , Edwards L D
Published: January 2020

The efficacy of the use of a single modified Thayer-Martin triplate versus that of three separate modified Thayer-Martin plates for the recovery of Neisseria gonorrhoeae from pharyngeal, anal, and genital sites were studied. A total of 98 males and 74 females who showed intracellular gram-negative diplococci on genital Gram stain were studied. Of 172 patients, 164 had gonorrhea at one or more sites. In the first group of 45 males and 21 females, a single swab from each site was used to inoculate randomly both a standard plate (100 mm in diameter) and one-third of a triplate. In the second group of 53 males and 53 females, two swabs were used for collection from each site. One of each pair of swabs was randomly inoculated onto a standard plate and a triplate. There was no significant difference between the results obtained by inoculation with single swabs and those obtained by inoculation with two swabs. There were no significant differences between the positivity rates obtained with the triplate and those obtained with three standard plates used at each site. Cost (39% that of three separate plates) and acceptance by clinic and laboratory personnel make the triplate method an economical, accurate, and effective triple-site screening system.
Immunoenzymatic absorption test for serodiagnosis of paracoccidioidomycosis
Authors: Mendes-Giannini M J , Camargo M E , Lacaz C S , Ferreira A W
Published: January 2020

A sensitive enzyme-linked immunosorbent assay is described for paracoccidioidomycosis serodiagnosis, with antigen represented by filtrates from Paracoccidioides brasiliensis cultures. Cross-reactivities were, however, observed with sera from patients with other mycoses such as histoplasmosis, lobomycosis, cryptococcosis, candidiasis, and sporotrichosis. These cross-reactions did not occur when we used as antigen Yarzabal E2 component prepared by affinity fractionation of the culture filtrates. Specific results could also be obtained with the whole filtrate antigen when sera were absorbed with Histoplasma capsulatum yeast and mycelial components. Besides a very high sensitivity (100% of the 69 cases of paracoccidioidomycosis tested), this assay showed no false-positive results for the 206 non-paracoccidioidomycosis sera studied, including those from patients with other mycotic diseases.
Two coagulase-variant forms of Staphylococcus aureus isolated from blood cultures
Authors: Fung J C , Kaplan M H , Hsieh H C , Stephens A , Tyburski M B , Tenenbaum M J
Published: January 2020

Two coagulase-variant forms of Staphylococcus aureus were isolated from blood cultures of a patient with infective endocarditis. The coagulase-positive isolate was hemolytic, whereas the coagulase-negative isolate was nonhemolytic. All other properties examined were identical in both strains. Since coagulase-negative S. aureus strains have been isolated from clinical specimens, laboratories should consider using a combination of other biological properties along with coagulase production for the identification of S. aureus.
Classification of Inoue-Melnick virus into three antigenic types
Authors: Inoue Y K , Nishibe Y , Melnick J L
Published: January 2020

Antigenic analysis of Inoue-Melnick virus, formerly called SMON virus, showed that isolates from patients with subacute myelo-optico-neuropathy (SMON) in Japan, thus far tested, belonged to a single antigenic type (type 1). However, Inoue-Melnick virus isolates obtained so far in the United States chiefly from patients with multiple sclerosis could be classified into three types: type 1, type 2, and an intermediate type. At the present state of knowledge, the virus has not been proven to be the causative agent of disease, hence its provisional designation as IMV.
Comparison of immediate and delayed inoculation of HEp-2 cells for isolation of respiratory syncytial virus
Authors: Bromberg K , Daidone B , Clarke L , Sierra M F
Published: January 2020

Immediate inoculation of HEp-2 cells is generally advocated for the isolation of respiratory syncytial virus. However, delayed laboratory inoculation of properly transported specimens obtained by aspiration of nasopharyngeal mucus provided an isolation rate similar to that obtained with immediate inoculation.
Vibrio fluvialis and Vibrio furnissii isolated from a stool sample of one patient
Authors: Hickman-Brenner F W , Brenner D J , Steigerwalt A G , Schreiber M , Holmberg S D , Baldy L M , Lewis C S , Pickens N M , Farmer J J
Published: January 2020

Vibrio fluvialis and Vibrio furnissii have been associated with diarrhea but have rarely been isolated in the United States. We received strains of V. fluvialis and V. furnissii that were isolated from the stool of a 1-month-old baby. A description of these two strains and the case history of the patient are given in this report.
Immunoblot analysis of Toxoplasma gondii antigens by human immunoglobulins G, M, and A antibodies at different stages of infection
Authors: Partanen P , Turunen H J , Paasivuo R T , Leinikki P O
Published: January 2020

The Toxoplasma gondii antigenic components eliciting the immunoglobulin G (IgG), IgM, and IgA antibody responses were studied by using follow-up sera from a laboratory worker who developed an acute glandular toxoplasmosis after an accidental infection with the protozoa. IgG toxoplasma antibodies reacted with multiple components over a wide molecular weight range from 6,000 to 150,000. In contrast, IgM toxoplasma antibodies reacted predominantly with polypeptides of 6, 25, and 35 kilodaltons, which might be useful in new diagnostic procedures. The general pattern of antigenic components in the IgA toxoplasma antibody response closely resembled that in the IgM response, even though some characteristic features were constantly observed. The possibility that the restricted IgM and IgA antibody responses relate to the pathogenetic events in human toxoplasmosis is considered.
Primary isolation and serial passage of hepatitis A virus strains in primate cell cultures
Authors: Binn L N , Lemon S M , Marchwicki R H , Redfield R R , Gates N L , Bancroft W H
Published: January 2020

Although several primate cell types have been reported to support replication of hepatitis A virus, optimal conditions for the isolation and production of quantities of virus have not been defined. We therefore examined seven different primate cell types for their ability to support replication of primate-passaged and wild-type virus as reflected by intracytoplasmic accumulation of viral antigen (direct immunofluorescence and radioimmunoassay) and propagation of cell culture-adapted virus. Of the cells tested, low-passage African green monkey kidney (AGMK) cells were most sensitive for initial isolation. Viral replication was documented after inoculation of AGMK cells with seven of nine hepatitis A virus antigen-positive fecal specimens (from seven epidemiologically distinct sources). With six inocula, virus was successfully passed in serial cultures. AGMK-adapted virus was readily propagated in continuous AGMK (BS-C-1) cells. The optimal temperature for the growth of virus in BS-C-1 cells was 35 degrees C. Viral release into supernatant fluids was documented in the absence of any cytopathic effect, and infectivity titers in supernatant fluids 21 days after inoculation (50% tissue culture infective does [TCID50], 10(6.0)/ml) equalled or exceeded those in the cell fraction (TCID50, 10(5.5)/ml). Cells maintained in serum-free media readily supported viral growth, with yields of virus (TCID50, 10(6.5)/ml) equal to or greater than those obtained with cells maintained in 2% fetal bovine serum.
Evaluation of the four-hour rapid 20E system for identification of members of the family Enterobacteriaceae
Authors: Izard D , Husson M O , Vincent P , Leclerc H , Monget D , Boeufgras J M
Published: January 2020

A study was conducted to compare the API Rapid 20E 4-h system (API System S.A., France; commercially available in the U.S.A. under the name DMS Rapid E System; DMS Laboratories, Darts Mill, Flemington, N.J.), the API 20E 18- to 24-h system (Analytab Products, Plainview, N.Y.), and a conventional media system to measure the ability of each to identify members of the family Enterobacteriaceae. Comparison tables rather than simple percentage agreement tables were generated to define the particular strengths and weaknesses of each system and to allow the laboratory to best use the data. The Rapid 20E compared quite favorably with conventional media. It yielded correct identifications with 95.9% of the isolates tested (API 20E, 98% identification rate). In 2.5% of the isolates, the Rapid 20E gave only genus identifications, and in 1.4% the organisms did not correspond to any key in the code book and could not be identified by the manufacturer's computer service. The ease of inoculation and the 4-h capability make the Rapid 20E system an extremely attractive development in the field of bacterial identification.
Trans-isolate medium: a new medium for primary culturing and transport of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae
Authors: Ajello G W , Feeley J C , Hayes P S , Reingold A L , Bolan G , Broome C V , Phillips C J
Published: January 2020

A diphasic medium, Trans-Isolate medium, was developed for the transport of primary cultures of cerebrospinal fluids from patients with bacterial meningitis. It consists of a charcoal-starch agar slant and soybean-casein digest-gelatin broth buffered at pH 7.2 with 0.1 M 3-(N-morpholino)propanesulfonic acid buffer. In the laboratory, this medium supported the growth and survival of stock cultures of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae for at least 3 months. Under field conditions in Africa, cerebrospinal fluids from patients suspected of having bacterial meningitis were inoculated directly onto plates of chocolate agar medium and into bottles of Trans-Isolate medium. An etiological agent was isolated from 52 spinal fluids by direct plating. After shipment to Atlanta, Ga., 2 to 4 weeks later, the same etiological agents were recovered from 38 bottles of Trans-Isolate medium.
Aminoglycoside resistance among blood culture isolates
Authors: Huovinen P , Grönroos P , Herva E , Katila M L , Klossner M L , Renkonen O V , Toivanen P
Published: January 2020

A total of 633 blood culture isolates were collected from 1981 to 1982 from seven major Finnish hospitals, including all university central hospitals. Susceptibility of the strains to gentamicin, tobramycin, amikacin, and netilmicin was determined by the Sensititre microtiter procedure. Resistance against any of these agents occurred in 1.3 to 6.5% of all strains studied. In the Turku University Central Hospital, an increased number of tobramycin- and gentamicin-resistant Staphylococcus epidermidis were found; the frequency of strains resistant to tobramycin was 57% and to gentamicin was 29% versus frequencies of 16 and 18%, respectively, which were observed in the other hospitals. An explanation for this might be a change in the use of aminoglycosides in the Turku University Central Hospital; within 3 years, 1979 to 1981, the consumption of tobramycin and amikacin had increased 330 and 290%, respectively, whereas the use of gentamicin had decreased to 24% of that in the beginning of the period. Resistance against tobramycin was mediated by enzymes APH(2")-AAC(6') and ANT(4').
Typing of Enterobacter spp. by bacteriocin susceptibility and its use in epidemiological analysis
Authors: Bauernfeind A , Petermüller C
Published: January 2020

Most clinical isolates of Enterobacter cloacae are bacteriocinogenic and susceptible to bacteriocins. Both rapidly diffusing, nonsedimentable, protease-susceptible and slowly diffusing, sedimentable, protease-resistant bacteriocins are produced. A practicable system was devised for epidemiological typing of E. cloacae isolates by their patterns of susceptibility to bacteriocins. A set of eight bacteriocin-producing strains was grown on tryptic soy agar plates for 16 h. After removal of the producer lawn, the isolates to be typed were inoculated on the agar media by a multipoint inoculator. After a second 16-h period of incubation, the strains were classified into bacteriocin types according to the patterns of growth inhibition. Typability of 134 clinical isolates was 96.3%. Only 11 (8.2%) of the isolates fell into the largest group. Repeat testing of isolates from the same patients within 2 months gave identical bacteriocin types. Other species of Enterobacter (E. agglomerans and E. aerogenes) are also typable by this method.
Selective enrichment broth culture for detection of Clostridium difficile and associated cytotoxin
Authors: Buchanan A G
Published: January 2020

A procedure was devised for routine examination of feces for Clostridium difficile with selective enrichment broth culture containing increased levels of carbohydrates and antibiotics to detect cytotoxin and volatile acids in broths inoculated with fecal samples. C. difficile was detected and identified with a rapidity comparable to that of conventional culture on selective cycloserine-cefoxitin fructose agar. Detection rates for C. difficile in inoculated broths (111/401 or 27%) were significantly higher than for culture on cycloserine-cefoxitin fructose agar (47/401 or 11%, P greater than 0.001). All fecal samples containing C. difficile and cytotoxin were correctly identified by the procedure. Isocaproic acid peak heights greater than 2 mm in selective enrichment broths inoculated with fecal samples indicated that C. difficile was present in the fecal sample examined. Of the positive specimens examined, 58% (64/111) produced peak heights greater than 10 mm. Peak heights less than 2 mm were not associated with C. difficile in the fecal sample. The investigated procedure provided a reliable alternative to the routine processing of feces for detecting C. difficile and associated cytotoxin in feces. Inoculated broths with isocaproic acid peak heights greater than 2 mm, after 24 to 48 h of incubation, and in which cytotoxin was detected, were subcultured to blood agar to obtain isolates of the organism as required. Broths which showed isocaproic acid peak heights less than 2 mm, and in which cytotoxin was not detected, were discarded as negative for C. difficile. The procedure was deemed potentially useful for epidemiological surveys of C. difficile.
Systematic investigation of enrichment media for wild-type Campylobacter jejuni strains
Authors: Barot M S , Bokkenheuser V D
Published: January 2020

Of the media examined, thioglycolate broth supplemented with 5% lysed sheep blood, Butzler antibiotic mixture, and 0.1% lauryl sulfate was the most sensitive enrichment medium for recovery of wild-type strains of Campylobacter jejuni from cecal contents of chickens and chicken livers. It allowed the retrieval of 1 CFU as did solid media but permitted the screening of 50-times larger volumes. Double-strength enrichment medium required 5 to 10 CFU for growth. Omission of lauryl sulfate reduced the sensitivity. Replacement of Butzler antibiotic mixture with Blaser antibiotic mixture increased overgrowth and, therefore, decreased retrieval of C. jejuni.
Isolation of immunogenic neuraminidases of human influenza viruses by a combination of genetic and biochemical procedures
Authors: Gallagher M , Bucher D J , Dourmashkin R , Davis J F , Rosenn G , Kilbourne E D
Published: January 2020

Neuraminidases were purified from reassortant viruses (H6N1 and H6N2) containing the two antigenic subtypes (N1 and N2) found in human influenza viruses. Surface glycoproteins were solubilized with octylglucoside, and the neuraminidase was isolated by chromatography on DEAE-Sephadex. Neuraminidase isolated by this technique coeluted with viral lipids and spontaneously formed liposomes on dialysis. The purified neuraminidase was immunogenic in rabbits, producing a significant antibody response at dose levels as low as 1 microgram.
Genetic relationship of lyme disease spirochetes to Borrelia, Treponema, and Leptospira spp
Authors: Hyde F W , Johnson R C
Published: January 2020

Genetic studies were performed on the following spirochetes: three Lyme disease spirochetes isolated from Ixodes ticks and from human spinal fluid; three species of North American borreliae; four species of Treponema; and two species of Leptospira. The mol% G+C values for Lyme disease spirochetes were 27.3 to 30.5%, similar to values of 28.0 to 30.5% for Borrelia species but different from the values of Leptospira or Treponema species which ranged from 35.3 to 53%. Lyme disease spirochetes represent a new species of Borrelia, with DNA homologies of 31 to 59% with the three North American strains of Borrelia studied. These studies also showed that Lyme disease spirochetes from three sources constituted a single species, with DNA homologies ranging from 76 to 100%. A high degree of relatedness was also seen between the three North American borreliae, with homology varying from 77 to 95%, indicating that these spirochetes represent a single species. Lyme disease spirochetes and Borrelia species exhibited almost no homology with Leptospira and Treponema species (0 to 2%). Plasmids were detected in the three Lyme disease spirochetes and in the three North American borreliae.
Identification and typing of herpes simplex virus by enzyme immunoassay with monoclonal antibodies
Authors: Frame B , Mahony J B , Balachandran N , Rawls W E , Chernesky M A
Published: January 2020

A double-antibody enzyme immunoassay was developed for the identification and typing of herpes simplex virus (HSV) by employing a polyclonal rabbit capture antiserum together with type-common and type 2-specific monoclonal antibodies as detectors. The test successfully identified 45 type I isolates and 30 type 2 isolates as HSVs. Compared with immunofluorescent staining and restriction endonuclease analysis, enzyme immunoassay correctly typed 45 type 1 and 30 type 2 HSV isolates. Enzyme immunoassay was 100% sensitive for identification of HSV as compared with cell culture and 100% specific for typing as compared with immunofluorescence and restriction endonuclease analysis. Electron microscopy analysis suggested that approximately 10(6) virus particles were required for the identification and typing of HSV by enzyme immunoassay.
Disseminated candidiasis caused by a sucrose-negative variant of Candida tropicalis
Authors: Ahearn D G , Lawrence J B
Published: January 2020

A patient suffering from leukemic transformation of a non-Hodgkin's lymphoma developed fatal disseminated candidiasis caused by a sucrose-negative variant of Candida tropicalis. The results of histopathological tests showed massive tissue invasion in many organs. The course of infection and the tissue morphology of the etiological agent were indistinguishable from those of C. albicans and typical C. tropicalis strains.
Serological analysis of five serotypes of Pasteurella multocida of rabbit origin by use of an enzyme-linked immunosorbent assay with lipopolysaccharide as antigen
Authors: Cary C J , Peter G K , Chrisp C E , Keren D F
Published: January 2020

The serological relationships among five Pasteurella multocida strains, representing five somatic serotypes most commonly isolated from rabbits (serotypes 1, 3, 4, 12, and 15), were studied with an enzyme-linked immunosorbent assay. Lipopolysaccharides from the five serotypes were used as antigens in the assay. Antisera against serotypes 1 and 15 reacted only with their homologous lipopolysaccharides. Significant cross-reactivity was found between serotypes 4 and 12. Serotype 3 antisera showed minimal reactivity with both homologous and heterologous lipopolysaccharides. The feasibility of detecting antibodies to each of several lipopolysaccharide antigens combined in the same assay cell well was demonstrated.
Comparison of latex agglutination and immunofluorescence for direct Lancefield grouping of streptococci from blood cultures
Authors: Shlaes D M , Toossi Z , Patel A
Published: January 2020

Simulated positive blood cultures with 84 known stock strains of streptococci were used to comparatively evaluate the direct identification of these organisms by fluorescein-tagged antibody staining (immunofluorescence [IF]) and latex agglutination (LA). IF was not evaluated for Lancefield group D strains (a total of 81 strains tested) and had 89% sensitivity and 91% specificity. IF was least sensitive for the identification of Lancefield group F, in which three of seven strains showed no fluorescence with the group F reagent. Since LA was more convenient and revealed comparable sensitivities and specificities on 84 simulated cultures, we tested this procedure using an additional 29 fresh positive clinical blood cultures, for a total of 113 cultures tested by this technique. Of 11 Streptococcus pneumoniae strains, 9 reacted with the LA group C reagent, a problem not observed with IF. However, all these strains were identified by a rapid modified bile solubility test. Of the 12 Streptococcus faecalis strains, 4 were falsely negative with the group D reagent, but all were correctly identified by a rapid litmus milk reduction test. Of 12 group A strains, 1 was not detected. Of all 113 strains tested by LA, eliminating S. faecalis and S. pneumoniae, the sensitivity and specificity were 97 and 98%, respectively. LA was simple and reliable in the rapid identification of streptococci from blood cultures and appeared to be preferable to IF. When LA is used, the group D reagent should not be used, and all samples reacting with the group C reagent should be tested by a modified rapid bile solubility test to exclude S. pneumoniae.
Monoclonal antibody that preferentially binds polylysine, polyarginine, and histones and selectively decorates nuclei and chromosomes
Authors: Morgan J L , Dennis D D
Published: January 2020

A monoclonal antibody, designated J-57, selectively and uniformly decorates the interphase nuclei and mitotic chromosomes of a variety of eucaryotic cells as determined by indirect immunofluorescence. As determined by enzyme-linked immunosorbent assay, however, this monoclonal antibody is not monospecific. It reacts weakly with cytochrome c, RNase A, and brain tubulin. By these tests monoclonal antibody J-57 has broad cross-reactivity similar to that of antisera directed against polylysine. The differential reactions of this monoclonal antibody suggest that it may be a useful immunohistochemical probe for nuclei and chromosomes in whole cells.
Susceptibility of hepatitis B virus to disinfectants or heat
Authors: Kobayashi H , Tsuzuki M , Koshimizu K , Toyama H , Yoshihara N , Shikata T , Abe K , Mizuno K , Otomo N , Oda T
Published: January 2020

Using direct chimpanzee inoculation as an assay method, we tested the abilities of the following chemical or physical treatments to inactivate hepatitis B virus in human plasma: 1% aqueous glutaraldehyde at 24 degrees C for 5 min, 0.1% aqueous glutaraldehyde at 24 degrees C for 5 min, 80% ethyl alcohol at 11 degrees C for 2 min, and heat at 98 degrees C for 2 min. All treatments were shown to be effective, indicating that the resistance level of the hepatitis B virus is not extreme.
Phenotypic factors correlated with the absence of virulence among gentamicin-resistant Pseudomonas aeruginosa strains
Authors: Clark R B , Janda J M , Bottone E J
Published: January 2020

Previous reports have suggested that clinical strains of gentamicin-resistant Pseudomonas aeruginosa (GRPA) generally produce only superficial infections (wounds, urinary tract infections) in contrast to their more invasive gentamicin-susceptible counterparts (GSPA). In view of this finding, a comparative study of a number of phenotypic properties of 20 GRPA and GSPA strains (10 isolates) was assessed to determine how closely related these two populations are and how their phenotypic properties might reflect virulence. GRPA isolates were found to be more adherent to buccal cells than their susceptible counterparts (P = 0.0001). Motility, however, was generally restricted in the GRPA population when compared with GSPA isolates (P = 0.02), although on the basis of zone diameters, some strains overlapped into the other group. Enzymatically, GSPA isolates produced significantly more lipase activity against C-14 lipids than GRPA strains (P = 0.04). No differences were recorded between the two populations in dye sensitivity or in their ability to grow on minimal media at 37 and 42 degrees C. Only 35% of the GRPA isolates agglutinated in 1 of the 17 monospecific antisera reactive for P. aeruginosa. The results of this study suggest that in vivo-generated GRPA strains possess phenotypic properties intermediate between those described for in vitro-derived GRPA isolates and their progenitor GSPA strains. The increased adherence of clinical GRPA isolates to buccal cells may explain their predilection to produce wound and urinary tract infections, whereas their lack of systemic dissemination may be partially due to decreased motility and to reduced lipase production.
Pulmonary disease due to Mycobacterium malmoense
Authors: Warren N G , Body B A , Silcox V A , Matthews J H
Published: January 2020

A case of pulmonary disease due to Mycobacterium malmoense was recently diagnosed in a 43-year-old man from Virginia. This organism was isolated from sputum and bronchial washings. This is the first case of documented human infection due to this organism in the United States
Filter radioimmunoassay, a method for large-scale serotyping of Neisseria meningitidis
Authors: de Marie S , Hoeijmakers J H , Poolman J T , Zanen H C
Published: January 2020

A simple and rapid filter radioimmunoassay method can be used to serotype meningococcal strains on a large scale. The technique consists of simultaneous inoculation of 96 strains on nitrocellulose filters. The resulting colonies can be processed in situ, by extraction and fixation, incubation with antibodies and 125I-labeled protein A, and, finally, autoradiography. Processing many filters simultaneously, one person can serotype thousands of meningococci in a week. Multiple filters with identical strain patterns can be stored after the fixation step for future screening. The use of monoclonal antibodies is essential; polyclonal antisera, even after extensive absorption, were not specific in this assay. When results from filter radioimmunoassay and Ouchterlony microprecipitation were compared for the serotyping of 201 Neisseria meningitidis strains for serotypes 2a and 2b, filter radioimmunoassay was sufficiently sensitive and specific to be useful in mass screening.
Infection of the olecranon bursa by Anthopsis deltoidea
Authors: Kwon-Chung K J , Droller D D
Published: January 2020

Anthopsis deltoidea was found to be the cause of an olecranon bursitis in a 79-year-old golfer. Serial histological sections of the olecranon bursa showed faintly stained, brown-walled, septate, hyphal elements in the centers of the necrotic debris. The combination of bursectomy and flucytosine treatment cured the infection.
Prevalence of Clostridium difficile and its cytotoxin in infants in Mexico
Authors: Torres J F , Cedillo R , Sánchez J , Dillman C , Giono S , Muñoz O
Published: January 2020

The incidence of Clostridium difficile and its cytotoxic activity were determined in the feces of 122 children under 1 year of age. Samples were obtained from children receiving antibiotics and with (52 cases) or without (26 cases) diarrhea, from children with diarrhea who did not receive antibiotics (22 cases), and from healthy children (22 cases). Isolation of C. difficile in feces from children in all groups was similar (mean 23.4%) except for the group with non-antibiotic-associated diarrhea (4.5%). In both groups of children receiving antibiotics, with or without diarrhea, the cytotoxin was detected in 7.6% of the cases. In the group with non-antibiotic-associated diarrhea, none of the samples was positive for cytotoxicity. In healthy children, cytotoxin was positive in 4.5% of the cases.
Pyomyositis caused by Yersinia enterocolitica
Authors: Brennessel D J , Robbins N , Hindman S
Published: January 2020

Yersinia enterocolitica is known to be associated with gastroenteritis in children and Reiter's syndrome in adults, but it has only rarely been the cause of primary soft tissue infections. A patient with diabetes mellitus developed a calf abscess, from which Y. enterocolitica was isolated in pure culture. Incision, drainage, and intravenous gentamicin therapy resulted in cure.
Use of a simple separation column in detection of immunoglobulin M antibody to Epstein-Barr virus
Authors: Sumaya C V , Ench Y , Carrillo M A , Pope R M
Published: January 2020

The use of a new commercial separation column on whole test serum improved the method for detecting immunoglobulin M antibody to Epstein-Barr virus. The efficiency of this product in absorbing interfering immunoglobulin G was similar to that of another absorption product, staphylococcal protein A, but it has the advantage of being stable without refrigeration.
Polycarbonate-coated microsticks as solid-phase carriers in an enzyme-linked immunosorbent assay for rubella antibody
Authors: Shekarchi I C , Tzan N , Sever J L , Madden D L
Published: January 2020

We evaluated the use of microsticks as solid-phase carriers in an enzyme-linked immunosorbent assay for rubella antibody. The microstick enzyme-linked immunosorbent assay was found to be equal in sensitivity to plate and disk enzyme-linked immunosorbent assays and presumably more sensitive than hemagglutination and immunofluorescence assays. The microstick as a solid-phase carrier offers advantages over both plate and bead carriers.
Laboratory evaluation of five assay methods for vancomycin: bioassay, high-pressure liquid chromatography, fluorescence polarization immunoassay, radioimmunoassay, and fluorescence immunoassay
Authors: Pfaller M A , Krogstad D J , Granich G G , Murray P R
Published: January 2020

We compared the precision and accuracy of five methods used to measure the concentration of vancomycin in serum: bioassay, high-pressure liquid chromatography, fluorescence polarization immunoassay (FPIA [TDX; Abbott Laboratories, North Chicago, Ill.]), radioimmunoassay (RIA), and fluorescence immunoassay. Based on an analysis of seven standards and of 106 patient samples, all five methods were accurate, and four (bioassay, high-pressure liquid chromatography, FPIA, and RIA) were also precise. The FPIA was the most precise and the fluorescence immunoassay was the least precise of the methods tested; intrarun coefficients of variation for these two methods were 0.9 to 3.0% versus 8.9 to 14.5%, and interrun coefficients of variation were 2.8 to 8.1% versus 12.2 to 16.2%, respectively. The RIA was inconvenient because it required an extra dilution of the specimen being tested and an additional (64 micrograms/ml) vancomycin standard for specimens with 32 to 64 micrograms of vancomycin per ml. Based on its rapid turnaround time and the stability of its standard curve, we believe that the FPIA is the best method currently available to quantitate vancomycin in the clinical laboratory.
Use of 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan as a primary medium for recovery of Pseudomonas aeruginosa from clinical specimens
Authors: Araj G F
Published: January 2020

Several concentrations of 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan (C-390), ranging from 5 to 100 micrograms/ml, were incorporated in brain heart infusion agar, MacConkey agar, and xylose-lysine-deoxycholate agar to evaluate the recovery of Pseudomonas aeruginosa from 538 sputum, 174 urine, and 22 stool samples. Seventy-six sputum samples containing P. aeruginosa grew this bacterium alone on brain heart infusion and MacConkey agars with a C-390 concentration of 25 micrograms/ml or greater. Other microorganisms present in these specimens grew only on media without C-390, and significantly less growth was observed on media with less than 20 micrograms of C-390 per ml. In few samples containing Klebsiella pneumoniae (3 of 30) and Serratia spp. (3 of 10), these organisms grew on all C-390 media and concentrations tested. The remaining sputum samples grew other bacteria and yeasts only on media without C-390. Brain heart infusion and MacConkey agars with C-390 were equally effective in recovering P. aeruginosa and suppressing the growth of a wide range of bacteria and yeasts from urine and stool samples. Xylose-lysine-deoxycholate agar with C-390 did not show a selective or suppressive advantage over xylose-lysine-deoxycholate agar alone for recovering P. aeruginosa from stool specimens. These results indicate that use of the correct medium and C-390 concentration would provide a suitable primary medium for inhibiting a wide range of bacteria and yeasts and would select the growth of P. aeruginosa from clinical specimens.
Latex agglutination test for detection of Clostridium difficile toxin in stool samples
Authors: Shahrabadi M S , Bryan L E , Gaffney D , Coderre S E , Gordon R , Pai C H
Published: January 2020

A total of 163 stool specimens were tested for detection of Clostridium difficile and its toxin by cytotoxicity assay with tissue culture, latex agglutination test, and isolation of the organism. From 33 specimens which were positive for toxin by cytotoxicity, 30 were positive by the latex agglutination test; the organism was isolated from 21. The total number of samples which were positive with the latex agglutination test was 44. The predictive value of a positive latex agglutination result relative to the cytotoxicity test was 68%, and the predictive value of a negative result was 97.5%. The specificity and sensitivity of the latex agglutination test relative to the cytotoxicity assay and the low cost and simple facilities required indicate that the latex agglutination test is a useful procedure for screening for C. difficile toxins, provided that positive latex results are confirmed by cytotoxicity assay.
Comparison of enzyme-linked immunosorbent and indirect hemagglutination assays for determining anthrax antibodies
Authors: Johnson-Winegar A
Published: January 2020

An enzyme-linked immunosorbent assay has been established to measure anthrax antibody titers. The protective antigen component of anthrax toxin was used as the capture antigen. Two types of conjugates (protein A-horseradish peroxidase and anti-human immunoglobulin G plus immunoglobulin A plus immunoglobulin M-horseradish peroxidase) were tested. Results from enzyme-linked immunosorbent assay testing were compared with those from indirect hemagglutination titers on serum from vaccinees. The overall trend of enzyme-linked immunosorbent assay and indirect hemagglutination titers was significant. The enzyme-linked immunosorbent assay offered speed, precision, and reduced cost per test.
Molecular epidemiology of Legionella pneumophila serogroup 1
Authors: van Ketel R J , ter Schegget J , Zanen H C
Published: January 2020

The DNA of patient and environmental isolates of Legionella pneumophila serogroup 1 was analyzed by restriction endonuclease cleavage. The electrophoretic patterns of the DNA digests of isolates from a group of patients with Legionnaires disease acquired in a hospital were indistinguishable from one another and were identical to the DNA pattern of a strain isolated from the hot water supply of the hospital. On the other hand, they were easily differentiated from strains isolated from patients and hot water supplies in other hospitals in the same city. The homogeneity of populations of L. pneumophila serogroup 1 colonizing plumbing systems was also investigated by DNA restriction endonuclease analysis in three hospitals. We distinguished two subtypes in one hospital; the two other hospitals had homogeneous populations. Restriction endonuclease digest analysis of L. pneumophila serogroup 1 DNA enables subtyping and appears to be a useful method for examining the epidemiology of outbreaks of Legionnaires disease.
Demonstration of Isospora belli by acid-fast stain in a patient with acquired immune deficiency syndrome
Authors: Ng E , Markell E K , Fleming R L , Fried M
Published: January 2020

Isospora belli, like Cryptosporidium sp., a sporozoan parasite of the small intestine, has been isolated from both immunodeficient and immunologically normal persons. In immunocompetent persons the infection may be asymptomatic; it frequently causes mild and self-limited diarrhea. The spectrum of the disease in persons with acquired immune deficiency syndrome is not well established. I. belli did not stain well with the methods usually used for intestinal protozoa and was difficult to see unstained. However, it stained well with the acid-fast technique used for Cryptosporidium sp. and was easily identified by such stains. A case of combined infection with Cryptosporidium sp. and I. belli is reported.
Detection of experimental Chlamydia trachomatis eye infection in conjunctival smears and in tissue culture by use of fluorescein-conjugated monoclonal antibody
Authors: Taylor H R , Agarwala N , Johnson S L
Published: January 2020

The immunofluorescent staining of conjunctival cytology smears with an antichlamydial monoclonal antibody is a new method for the detection of chlamydial eye infection. Using a monkey model of chlamydial eye infection, we compared the monoclonal antibody staining cytology with the established methods of Giemsa-stained cytology and chlamydial culture with cycloheximide-treated McCoy cells. Compared with either culture or monoclonal antibody-stained cytology, Giemsa cytology had a low sensitivity (30 and 24%, respectively) but was highly specific (100% in each case). Compared with culture, monoclonal antibody-stained cytology was highly sensitive (94%) but appeared to lack specificity (70%). However, examination of the changes in identification rates with time suggested that monoclonal antibody-stained cytology may be even more sensitive than culture and detect lower levels of infection. This study showed that monoclonal antibody-stained cytology of conjunctival smears might be a rapid, efficient, and inexpensive method of diagnosing ocular chlamydial infection.
Disk diffusion testing, quality control guidelines, and antimicrobial spectrum of HR810, a fourth-generation cephalosporin, in clinical microbiology laboratories
Authors: Jones R N , Thornsberry C , Barry A L , Ayers L , Brown S , Daniel J , Fuchs P C , Gavan T L , Gerlach E H , Matsen J M
Published: January 2020

HR810 is a new, very broad-spectrum cephalosporin with significant activity against members of the family Enterobacteriaceae, pseudomonads, gram-positive cocci, and anaerobes that is generally greater than the third-generation cephalosporins (99.6% of 4,128 clinical facultative enteric isolates were inhibited by less than or equal to 8.0 micrograms of HR810 per ml). Tests and statistical methods to establish in vitro antimicrobial susceptibility test criteria favor tentative breakpoints of greater than or equal to 18 mm (less than or equal to 8.0 micrograms/ml) as susceptible and less than or equal to 14 mm (greater than or equal to 32 micrograms/ml) as resistant. This provides a 93.7 to 98.3% absolute interpretive accuracy. Several preliminary ranges for zone sizes obtained with quality control organisms are proposed for the 30-micrograms HR810 disk diffusion test used during the clinical trials.
Comparison of lysis-centrifugation with a biphasic blood culture medium for the recovery of aerobic and facultatively anaerobic bacteria
Authors: Henry N K , Grewell C M , Van Grevenhof P E , Ilstrup D M , Washington J A
Published: January 2020

The Du Pont Isolator tube and Roche Septi-Chek blood culture bottle employ solid media which facilitate the removal of bacteria from static or cidal substances in blood to increase recovery and decrease detection time. In a comparison of 11,567 blood culture sets, the Isolator tube and vented Roche Septi-Chek bottle were positive for 533 (80%) and 494 (74%) of the aerobic and facultatively anaerobic organisms recovered, respectively. This difference was not significant. A significant difference was found in the overall detection time. The Isolator tube recovered the bacteria ca. 1 day earlier. The earlier detection time was most notable with Staphylococcus aureus, viridans streptococci, and Pseudomonas aeruginosa. Among the 355 bacteremic episodes analyzed by a computer program, the Isolator tube was responsible more often for the first report of bacteremia in a given patient. Both systems performed well for the recovery of aerobic and facultatively anaerobic bacteria, but it is recommended that either be used in combination with an unvented broth-containing bottle.
Rapid recognition of methicillin-resistant Staphylococcus aureus by use of automated test systems
Authors: Jorgensen J H , Redding J , Johnson J E , Holloway V , Almeida R J
Published: January 2020

The ability to rapidly recognize methicillin-resistant Staphylococcus aureus by use of two automated instrument systems, the MS-2 system (Abbott Laboratories, Diagnostics Division, Irving, Tex.) and the AutoMicrobic system (Vitek Systems, Hazelwood, Mo.), was evaluated on a collection of 95 methicillin-resistant S. aureus isolates recovered from at least six geographical areas of the United States. Isolates were simultaneously tested with both systems, and the results were compared with MIC tests performed by the National Committee for Clinical Laboratory Standards agar dilution method. Methicillin-resistant S. aureus isolates were defined as those with a methicillin MIC greater than or equal to 8 micrograms/ml by the reference procedure. Overall, with the AutoMicrobic system, 94.7% of 95 methicillin-resistant S. aureus isolates were detected, and with the MS-2 system, 91.6% of the isolates were detected. Isolates with methicillin MICs greater than or equal to 32 micrograms/ml were readily detected with both systems (41 of 42 isolates). Of 53 isolates from three locales with methicillin MICs of 8 or 16 micrograms/ml, 90.6% (48) were detected by the AutoMicrobic system, whereas 86.8% (46) were detected by the MS-2 system. A program update which has been added to the MS-2 system prints a warning message indicating possible methicillin-resistant S. aureus with isolates which demonstrate multiple antibiotic resistance (greater than or equal to four drugs other than methicillin). This warning message would have provided presumptive recognition of six of eight isolates with discrepant results for methicillin by the MS-2 system.
Group B streptococcal colonization patterns in mothers and their infants
Authors: Jones D E , Kanarek K S , Lim D V
Published: January 2020

Maternity patients and their newborn infants were cultured for group B streptococci (GBS) at Tampa General Hospital, Tampa, Fla., from September 1982 to May 1983. Culture swabs were placed into Lim Group B Strep Broth (GIBCO Laboratories, Madison, Wis.) and quantitated for GBS. A strong correlation was found between the numbers of GBS in the maternal vagina and the infant rectum. Infants symptomatic for early-onset GBS disease were delivered by mothers heavily colonized (greater than or equal to 3 X 10(4) GBS per swab) at the vagina. Such mothers were identified as GBS carriers by slide coagglutination and latex agglutination after their broth cultures had been incubated for 5 h. These data indicate that maternity patients at high risk of delivering infants heavily colonized with GBS and potentially symptomatic for early-onset GBS disease can be rapidly and selectively identified.
Latex immunoassay for rapid detection of rotavirus
Authors: Hughes J H , Tuomari A V , Mann D R , Hamparian V V
Published: January 2020

A latex agglutination (LA) test was evaluated for the detection of human rotaviruses in stool specimens. Both antiserum and immunoglobulin G (IgG)-sensitized latex particles were used, with IgG-coated beads being more sensitive for human rotavirus antigen detection. Latex beads sensitized with anti-simian-SA-11 IgG were stable for at least 8 months when stored at 4 degrees C. The sensitivity of the test was compared with that of the Rotazyme (Abbott Laboratories, Diagnostics Div., North Chicago, Ill.) test. The least number of particles detected was 9.0 X 10(5) particles by the LA test versus 4.5 X 10(5) particles by the Rotazyme test. When 10 stool specimens were serially diluted for antigen endpoint determinations, the geometric mean titer by the LA test was 592 versus 1,280 by the Rotazyme test. Forty-three stool samples positive by the Rotazyme test were all positive by the LA test, and no false negative results were detected. Unconfirmed false positive reactions ranged between 8 and 24%. The LA test for rotavirus antigen detection is direct, easy to perform, sensitive, quick, and may have application for use in diagnostic laboratories, emergency rooms, and physician's offices.
Differentiation of Campylobacter species by protein banding patterns in polyacrylamide slab gels
Authors: Ferguson D A , Lambe D W
Published: January 2020

Soluble protein extracts of 37 catalase-positive strains of Campylobacter species were examined by polyacrylamide slab gel electrophoresis (PAGE). Electrophoretic banding patterns showed good correlation with biochemical tests and with available DNA homology data in distinguishing species of Campylobacter but did not differentiate subspecies or biotypes. PAGE patterns indicated that Campylobacter coli is a distinct species. Furthermore, the PAGE patterns indicated that C. jejuni and nalidixic acid-resistant thermophilic Campylobacter species (C. laridis) are each distinct species. The protein banding patterns of C. fetus subsp. venerealis and C. fetus subsp. fetus strains were distinctly different from those of the three thermophilic species.