bluuebunny/crossref_metadata_2025 huggingface dataset:
Method for avoiding false-positive results occurring in immunoglobulin M enzyme-linked immunosorbent assays due to presence of both rheumatoid factor and antinuclear antibodies
Authors: Naot Y , Barnett E V , Remington J S
Published: January 2020

In a double-sandwich immunoglobulin M (IgM) enzyme-linked immunosorbent assay recently developed for the detection of IgM antibodies to Toxoplasma, the presence of either rheumatoid factor or antinuclear antibodies did not cause false-positive results. We recently noted, however, that in certain sera containing both rheumatoid factor and antinuclear antibodies, false-positive results do occur. In experiments to define the nature of the cross-reaction, these false-positive results were not found to be a consequence of interactions of the sera with Toxoplasma antigens, but rather were due to interactions of rheumatoid factor-antinuclear antibodies with the Fc portion of IgG antibodies used for the enzyme conjugate. This was avoided when the F(ab')2 fragment of IgG was used for the conjugate. The use of such F(ab')2 conjugates did not affect the sensitivity and, thereby, the usefulness of the double-sandwich IgM enzyme-linked immunosorbent assay for the diagnosis of acute acquired or congenital Toxoplasma infections. We concluded that F(ab')2 fragments of IgG antibodies can be used as enzyme conjugates to avoid false-positive results in sera positive for both rheumatoid factor and antinuclear antibodies in either the conventional test or in our double-sandwich IgM enzyme-linked immunosorbent assay.
Comparison of the Phadebact Gonococcus Test with the rapid fermentation method
Authors: Futrovsky S L , Gaydos C A , Keiser J
Published: January 2020

The Phadebact Gonococcus Test (Pharmacia Diagnostics, Piscataway, N.J.), a coagglutination technique, was compared with the rapid fermentation method of Kellogg and Turner (D. S. Kellogg, Jr., and E. M. Turner, Appl. Microbiol. 25: 550--552, 1973). A total of 93 organisms isolated on Martin-Lewis media were determined to be Neisseria gonorrhoeae based on the following criteria: presence of gram-negative diplococci, oxidase positivity, and appropriate reaction in the rapid fermentation method. These 93 isolates were then serologically tested with the Phadebact test. The direct method was attempted on the first 46 N. gonorrhoeae isolates. Difficulty in interpreting results was encountered in 39%. Thereafter, the alternate method of boiling was instituted on an additional 47 N. gonorrhoeae isolates, with 2 isolates producing noninterpretable results. All 93 isolates were frozen for a maximum of 2 months in skim milk at -25 degrees C. These isolates were thawed and retyped with the alternate boiling procedure, with 97% being confirmed as N. gonorrhoeae. In addition, 33 Neisseria meningitidis isolates, 14 Neisseria species, and 7 Moraxella species were tested with similar techniques. No positive reactions were observed. A cost effectiveness study using 5, 10, and 20 microliters of the gonococcal reagent was undertaken to reduce the cost of the test. When 10 and 20 microliters of reagent were used, no difficulty was encountered in interpreting the reaction. The coagglutination technique was difficult to read when 5 microliters of reagent was used.
Hydrolytic enzymes of anaerobic bacteria isolated from human infections
Authors: Steffen E K , Hentges D J
Published: January 2020

Thirty-three strains of anaerobic bacteria isolated from human clinical specimens were examined for the presence of heparinase, hyaluronidase, chondroitin sulfatase, gelatinase, collagenase, fibrinolysin, lecithinase, and lipase activities. Pronounced heparinase activity was limited to species of the genus Bacteroides. A number of species of the genera Bacteroides and Clostridium produced hyaluronidase and chondroitin sulfatase. Gelatinase, collagenase, and fibrinolysin activities were encountered in isolates of the genera Bacteroides, Clostridium, and Peptostreptococcus. All strains capable of degrading collagen also hydrolyzed other protein substrates. Lipolytic activity was minimal among these anaerobic bacteria. No specific hydrolytic activity was consistently associated with the isolates.
Rapid presumptive identification of Citrobacter diversus as an aid in controlling infections
Authors: Anderson R L , Graham D R , Dixon R E
Published: January 2020

During an outbreak of neonatal meningitis caused by kanamycin-resistant Citrobacter diversus, a field procedure for presumptive identification of the organism was evaluated, and using it resulted in the early recognition of patients colonized by the epidemic strain of C. diversus. Rectal and nasopharyngeal specimens were plated and incubated on MacConkey agar containing 10 micrograms of kanamycin per ml. After 18 h of incubation, lactose-nonfermenting colonies present on the selective medium were picked and identified 8 h later with the following biochemical tests: indole, adonitol, dulcitol, citrate, and triple sugar iron agar. This presumptive identification of C. diversus was later confirmed for all cases by conventional testing with a complete set of biochemicals. In 1 week, 253 specimens from patients were processed with this technique, and 49 strains of C. diversus were identified. The rapid identification of C. diversus and transfer of carriers into appropriate cohorts resulted in a 64% reduction in the prevalence of colonization.
Thiamine pyrophosphate (cocarboxylase) as a growth factor for Haemophilus somnus
Authors: Asmussen M D , Baugh C L
Published: January 2020

The effect of a commercially available, chemically defined enrichment (Iso-VitaleX; BBL Microbiology Systems, Cockeysville, Md.) on the growth of 10 strains of Haemophilus somnus was studied. A 6- to 10-fold increase in growth, as measured turbidimetrically, was observed when Iso VitaleX was added to a basal medium of brain heart infusion broth to a final concentration of 1% (vol/vol). Thiamine pyrophosphate (cocarboxylase), a constituent component of Iso VitaleX, was found to be the only growth-promoting factor, and it could be used as a substitute for Iso VitaleX. An equimolar concentration (2.2 microM) of thiamine monophosphate promoted growth equal to that of thiamine pyrophosphate. Thiamine was nonstimulatory for all 10 strains tested. When alkaline thermal-treated brain heart infusion broth was used as the basal medium, 7 of the 10 strains had an absolute requirement for thiamine monophosphate or thiamine pyrophosphate. The three remaining strains showed minimal growth when thiamine was added to this basal medium; however, excellent growth was observed when thiamine monophosphate or thiamine pyrophosphate was utilized. Factor X (hemin) was found to further enhance the growth when concentrations of 5 to 10 micrograms/ml were coupled with thiamine pyrophosphate. No increase in growth was observed when factor V (nicotinamide adenine dinucleotide) was coupled with thiamine pyrophosphate. This is the first report of a growth factor requirement for H. somnus.
FK phage for differentiating the classical and El T or groups of Vibrio cholerae
Authors: Takeya K , Otohuji T , Tokiwa H
Published: January 2020

A new vibrio-infecting phage (FK phage) isolated from sewage lysed all strains of Vibrio cholerae biovar cholerae, whereas all strains of V. cholerae biovar El Tor were resistant to it. FK phage was entirely different from Mukerjee group IV phage in morphology and antigenicity. In addition to group IV phage, the use of FK phage will be useful in the examination and typing of V. cholerae.
Enzymatic characterization of some oral and nonoral gram-negative bacteria with the API ZYM system
Authors: Slots J
Published: January 2020

The API ZYM system (Analytab Products, Plainview, N.Y.), containing 19 chromogenic substrates, was utilized semiquantitatively to detect extracellular acid and alkaline phosphatases, aminopeptidases, proteases, esterase-lipase, phosphoamidase, and glycosidases in 128 oral and nonoral isolates of black-pigmented Bacteroides, Actinobacillus, Haemophilus aphrophilus, Capnocytophaga, Fusobacterium nucleatum, Wolinella recta, and Veillonella parvula. In the black-pigmented Bacteroides group of organisms, a strong trypsin reaction was present in Bacteroides gingivalis (oral species) but not in Bacteroides asaccharolyticus (nonoral species). Bacteroides melaninogenicus subsp. melaninogenicus, in contrast to Bacteroides melaninogenicus subsp. intermedius, exhibited strong N-acetyl-beta-glucosaminidase activity. H. aphrophilus produced beta-galactosidase and alpha-glucosidase, but the closely related Actinobacillus actinomycetemcomitans did not. Capnocytophaga was distinct with respect to strong aminopeptidase reactions. This study showed that a wide range of enzymes which have the potential of causing tissue injury and inflammation can be elaborated from major oral gram-negative species. Also, the API ZYM system appears to be a valuable adjunct to traditional biochemical testing in identifying oral gram-negative species.
Colorimetric method for rapid determination of bacteriuria
Authors: Wallis C , Melnick J L , Longoria C J
Published: January 2020

An inexpensive, rapid, and simple colorimetric test for detection of bacteriuria is described. This test does not require bacterial growth and has the marked advantage of being able to quantify bacteria, even when the organisms are present in the urine of bacteriuric patients who are being treated with antibiotics. The test is carried out with 1 ml of urine, which is processed through a 10-mm-diameter filter than entraps the bacteria on its surface. Safranine dye is passed through the filter to stain the bacteria and the filter fibers. A decolorizer, which removes the dye from the filter fibers but not from the bacteria, is then passed through the filter. If there are greater than or equal to 10(5) colony-forming units of bacteria per ml in the sample, the 10-mm filter disk manifests a pink to red color. If there are less than 10(5)colony-forming units of bacteria per ml, the filter disk remains white or becomes slightly yellow. The entire procedure has been adapted to a semiautomated instrument and the time required per test is less than 1 min. The results obtained on the test card are a permanent record to be filled with the patient's chart. The bacteria can be quickly classified as gram positive or gram negative by selective staining of a second milliliter of urine on the filter. Of 441 urine specimens tested, 430 (98%) were correctly classified as containing more or less than 10(5) colony-forming units per ml. A total of 62 urine specimens were positive by bacterial plating (greater than or equal to 10(5) colony-forming units per ml), and 59 were positive by the colorimetric test. Eight false-positives (colorimetric test positive, plate counts less than 10(5)) were encountered in patients (bacteriuric) being prescribed antibiotics. Removal of the antibiotics from these urine specimens, with subsequent replating of the samples, indicated the presence of greater than or equal to 10(5) colony-forming units of bacteria per ml in three representative cases tested, indicating that the results of the colorimetric tests were not false-positives but that the plate counts were low because of the inhibition of bacterial growth by the residual antibiotics present in the urine.
Plasmid-encoded lysine decarboxylation in Proteus morganii
Authors: Cornelis G , Van Bouchaute M , Wauters G
Published: January 2020

As a rule, Proteus morganii does not decarboxylate lysine. However, lysine-positive P. morganii strains have been recently described. We suspected a plasmid origin for this atypical character, and we analyzed 14 strains to study this question. Among these strains, 8 yielded lysine-negative segregants after acridine orange or ethidium bromide treatment, and 10 transferred their lysine-positive character to a recipient P. morganii strain. All of the 14 strains analyzed at least segregated or conjugated. Three lysine-positive transconjugants, in turn, segregated lysine-negative variants after ethidium bromide treatment. The eight wild-type lysine-positive strains that segregated lysine-negative subclones contained a large (35 to 45 megadaltons) plasmid detectable by agarose gel electrophoresis. Similarly, the 10 lysine-positive transconjugants contained a plasmid of the same size, whereas P. morganii 1000, the recipient strain, did not contain any detectable plasmid. The large plasmid clearly disappeared in 9 of 11 lysine-negative segregants analyzed. It is concluded that the lysine-positive character of these P. morganii strains is plasmid encoded, and the taxonomical implications are discussed.
Evaluation of media for isolation of Campylobacter fetus subsp. jejuni from fecal specimens
Authors: Gilchrist M J , Grewell C M , Washington J A
Published: January 2020

Isolation rates of Campylobacter fetus subsp. jejuni from human fecal specimens were equivalent after broth enrichment (thioglycolate medium containing antibiotics) and direct inoculation on two brucella blood agar media containing ferrous sulfate, sodium metabisulfite, and sodium pyruvate, identical concentrations of vancomycin and trimethoprim, and different concentrations of polymyxin B and cephalothin. Studies with clinical isolates of C. fetus subsp. jejuni demonstrated temperature-dependent activity of polymyxin E (colistin) and substantial inhibition of growth on Thayer-Martin and Martin-Lewis media.
Clinical comparison of ethyl acetate and diethyl ether in the formalin-ether sedimentation technique
Authors: Erdman D D
Published: January 2020

A substitute for the volatile solvent diethyl ether has been actively sought for the Formalin-ether sedimentation technique. Ethyl acetate has recently been shown to be a comparable substitute. In an effort to verify these findings and evaluate ethyl acetate under clinical conditions, comparison studies with 62 fresh human stool specimens were performed. Parallel concentrates with diethyl ether and ethyl acetate were prepared for each specimen, and the quantity and appearance of recovered parasite species were determined. Ethyl acetate was found comparable to diethyl ether in the quantitative recovery of parasite eggs, cysts, and larvae; no distortion or alteration of parasite morphology was observed with either solvent. More care was required, however, to completely remove the interface plugs of ethyl acetate and prevent their remixing the concentrate sediment. In addition, wet mounts prepared from ethyl acetate concentrates were occasionally obscured by liquid bubbles probably composed of remaining insoluble ethyl acetate. Clinical laboratories considering substituting ethyl acetate for diethyl ether in the Formalin-ether sedimentation technique should be aware of these problems and take the appropriate precautions to avoid them.
Comparison of new triton X-100- and tween-ether-treated split-treated vaccines in children
Authors: Gross P A , Ennis F A , Gaerlan P F , Denning C R , Setia U , Davis W J , Bisberg D S
Published: January 2020

Split-product vaccines (SPVs) combine the desirable properties of no systemic reactogenicity and adequate immunogenicity when two doses are given. We compared a new Triton X-100 SPV (Connaught Laboratories, Inc.) with the commercially available Tween-ether SPV (Parke-Davis & Co.) in 76 children and young adults 2 to 25 years old; there were 39 and 37, respectively, in each vaccine group. Both vaccines contained influenza A/Brazil/78, A/Texas/77, and B/Hong Kong/72 (7 microgram of hemagglutinin for each strain); two doses were administered 1 month apart. Among persons seronegative by the hemagglutination inhibition test, the geometric mean antibody titers rose to approximately 100 after the first vaccination for influenza A/Brazil/78 and A/Texas/77. For B/Hong Kong/72, however, seronegative recipients developed lower geometric mean titers of approximately 32 after one immunization. Against the new B/Singapore/79 strain neither SPV stimulated adequate cross-reacting hemagglutination inhibition antibody (geometric mean titers of approximately 10). In conclusion, the new Triton X-100 SPV appears to be comparable to the ether-treated SPV in primed subjects. Further studies in unprimed children should be done to confirm this impression. In addition, it would be advisable to study other dosage regimens in unprimed children with these SPVs.
Lack of requirement for blind subcultures of BACTEC blood culture media
Authors: McLaughlin J C , Evers J L , Officer J L
Published: January 2020

To determine the need for blind subculturing of BACTEC (Johnston Laboratories, Cockeysville, Md.) blood culture media, we compared results of radiometric readings, visual inspection, and blind subculturing for nearly 7,500 blood specimens. Visual inspection and radiometric testing were performed on day 1 through 7, and blind subcultures were made on day 3. In the first phase of the study, 402 of 3,896 aerobic bottles were positive by radiometric testing (growth index, greater than 25), visual inspection, or subculturing. Only six bottles were radiometrically negative but subculture positive on day 3. The second phase of the study was designed to determine if aerobic bottles eventually became radiometrically positive in those cases in which they were radiometrically negative but subculture positive on day 3. Two bottles were subculture positive but never gave a growth index of greater than or equal to 25 by day 7. One yielded Staphylococcus epidermidis, and one yielded viridans, Streptococcus sp. A total of 35 anaerobic organisms were isolated from 3,896 blood specimens. All of these anaerobes were detected by both radiometric testing and subculturing. We examined a total of 14,972 blood culture bottles. Twenty-nine bottles considered negative by visual inspection or radiometric readings were found to be positive by subculturing. Fifteen of these were shown, by chart review, to contain contaminants. Organisms in the other negative bottles would not have gone undetected because companion bottles from the same patients were radiometrically or visually positive. We concluded that it is necessary to perform blind subcultures of BACTEC 7B and 8B blood culture bottles.
Effect of moisture content of the medium on colony morphology of Campylobacter fetus subsp. jejuni
Authors: Buck G E , Kelly M T
Published: January 2020

Reduction in the moisture content of the medium produced a profound effect on the colony morphology of Campylobacter fetus subsp. jejuni. Fresh medium produced flat, grayfish, spreading colonies with an irregular shape and variety appearance. Plates that were incubated at 30 degrees C for 48 h produced round, convex, butyrous colonies with an entire edge. Plates incubated at 30 degrees C for 24 h before inoculation produced colonies of an intermediate nature; they were round and raised, but not convex, and slightly water or mucoid in nature. This marked effect produced by moisture content of the medium was reproducible and may account for the variation in colonies observed by other investigators.
Recognition of a second serogroup of Legionella longbeachae
Authors: Bibb W F , Sorg R J , Thomason B M , Hicklin M D , Steigerwalt A G , Brenner D J , Wulf M R
Published: January 2020

A strain of Legionella longbeachae (Tucker 1) that was isolated from the postmortem lung tissue of a pneumonia patient was serologically distinct from four other strains of L. longbeachae. The recognition of a second serogroup of L. longbeachae represents the first reported instance of serogroup diversity within a species of Legionella other than L. pneumophila. The disease caused by the Tucker 1 strain does not seem to be readily distinguishable from that of pneumonia caused by other legionellae.
Quantitative evaluation of variation in composition of the streptozyme agglutination reagent for detection of antibodies to group A streptococcal extracellular antigens
Authors: Kaplan E L , Kunde C
Published: January 2020

The antibody titers of 90 acute- and convalescent-phase sera from 30 patients (mean age, 15.0 years) were studied with five different lots of Streptozyme reagent to determine whether variation exists among lots of reagent and whether this can affect clinical interpretation of antibody titers. Each serum was tested simultaneously with each of the five lots of reagent. Lot-to-lot variation in the reagent resulted in a significant difference (greater than 2 dilution increments) in antibody titer for 61 (68%) of the 90 sera tested. In addition, differences among the five lots also resulted in variation in determining whether a significant rise in titer occurred from the acute-phase to the convalescent serum for a given patient. These data indicate a need for precise identification and quantitation of the streptococcal antigens coated onto erythrocytes used for the Streptozyme test reagent.
Selenomonas sputigena
Septicemia
Authors: McCarthy Laurence R. , Carlson James R.
Published: January 2020

Selenomonas sputigena
is a curved motile anaerobic gram-negative rod that is a part of the normal upper respiratory tract of man. We present a case of speticemia associated with this organism and believe it to be the first such report describing systemic disease.
Cellular Fatty Acid Composition of
Legionella longbeachae
sp. nov
Authors: Moss C. Wayne , Karr D. E. , Dees S. B.
Published: January 2020

The cellular fatty acid composition of
Legionella longbeachae
was determined by gas-liquid chromatography. As in other
Legionella
species, the fatty acids of this new species are characterized by relatively large amounts of branched-chain acids.
Isolation of chlorhexidine-resistant Pseudomonas aeruginosa from clinical lesions
Authors: Nakahara H , Kozukue H
Published: January 2020

The chlorhexidine resistance of 317 strains of Pseudomonas aeruginosa isolated from hospital patients was determined. The distribution pattern of their susceptibility to chlorhexidine clearly revealed two peaks, and the frequency of resistance to chlorhexidine was 84.2%.
Mutanolysin enhancement of serogrouping of single colonies of streptococci
Authors: Calandra G B , Henson C L
Published: January 2020

Single colonies of beta-hemolytic streptococci could be grouped by antibody-coated latex bead agglutination or coagglutination with staphylococci coated with antibody after incubation of the colonies with mutanolysin. This simple and quick procedure provided an enzymatic means of enhancing the sensitivity of tests such as Phadebact and SeroSTAT.
Evaluation of the rapid penicillinase paper strip test for detection of beta-lactamase
Authors: Oberhofer T R , Towle D W
Published: January 2020

The penicillin-starch paper strip method was compared with the acidometric and iodometric methods for assaying beta-lactamase production, using fresh isolates of clinically important bacteria. Results obtained by the three methods were compared for rapidity, accuracy, and stability of reagents. Of the 210 isolates tested by the paper strip method, 301 isolates tested by the acidometric method, and 117 isolates tested by the iodometric method, all were in perfect agreement with the disk diffusion susceptibility test except one strain each of Haemophilus influenzae, Staphylococcus aureus, and Staphylococcus epidermidis. The H. influenzae isolate was penicillin resistant and failed to give a positive test for beta-lactamase in all three tests. The staphylococci (intermediate and resistant in susceptibility, respectively) failed to give a positive test for beta-lactamase with the iodometric method. The results of the paper strip method, in which 3,241 strains representing nine species of bacteria were used, correlated completely with disk susceptibility tests except for 2 and 69 strains, respectively, of penicillin-resistant, beta-lactamase-negative H. influenzae and H. parainfluenzae. The results of this study indicate that the paper strip method is accurate, simple to perform, extremely economical, and uses materials that are stable when stored frozen. It is eminently suitable for routine laboratory use.
Differentiation of Haemophilus spp. in Respiratory isolate cultures by an indole spot test
Authors: Welch D F , Ahlin P A , Matsen J M
Published: January 2020

Indole spot tests using isolated, nonhemolytic colonies of Haemophilus species were positive for 90 of 151 (60%) respiratory isolates of Haemophilus influenzae, whereas 67 to 72 (93%) isolates of H. influenzae from cerebrospinal fluid and blood specimens were indole positive. Only 4 of 117 (3%) Haemophilus parainfluenzae isolates were positive for indole spot tests. Thus, indole-positive, nonhemolytic Haemophilus isolates in respiratory cultures can be presumptively identified as H. influenzae.
Typing of herpes simplex virus with type-specific human immunoglobulin M in an indirect immunofluorescence assay
Authors: Spruance S L , Golden C A , Kern E R , Katz M E , Chow F
Published: January 2020

Sera from nine individuals with suspected primary herpes simplex virus type 1 (HSV-1) or type 2 (HSV-2) infection were screened to identify those containing HSV type-specific immunoglobulin M (IgM). Selected sera were then utilized in an IgM-specific indirect immunofluorescent-antibody HSV-typing assay (patent pending). To evaluate the procedure, 29 HSV isolates were grown in cultures of continuous human amnion cells, fixed, and used as substrates for indirect immunofluorescence. Determination of virus type was based on intensity of fluorescence of the substrate with HSV-1- and HSV-2-specific antisera after staining with fluorescein-conjugated anti-human IgM. Typing of the isolates by restriction endonuclease digestion showed that of 29, 18 were HSV-2 and 11 were HSV-1. Results by IgM-specific indirect immunofluorescent-antibody assay were identical to those by restriction endonuclease digestion for 27 of the isolates; 2 isolates failed to replicate adequately in the test cells. The IgM-specific indirect immunofluorescent-antibody procedure appears to be a simple, rapid, and accurate technique which could be of use to clinical virology laboratories.
Evaluation of the latex slide agglutination test for identification of Staphylococcus aureus
Authors: Myrick B A , Ellner P D
Published: January 2020

This study evaluated the reliability of the latex slide agglutination test for identifying Staphylococcus aureus. A total of 806 clinical isolates of staphylococci were tested for latex agglutination, clumping factor, and free coagulase. Positive latex tests occurred in 98.3% of coagulase-positive strains, whereas 99.6% of coagulase-negative strains gave negative latex tests. It is concluded that in most instances, the latex slide agglutination test is a reliable method for identifying S. aureus in the clinical laboratory.
Legionella jordanis: a new species of Legionella isolated from water and sewage
Authors: Cherry W B , Gorman G W , Orrison L H , Moss C W , Steigerwalt A G , Wilkinson H W , Johnson S E , McKinney R M , Brenner D J
Published: January 2020

Legionella jordanis sp. nov., as found in two cultures, is described. One isolate was from river water in Indiana and the other isolate was from sewage in DeKalb County, Ga. The former is the type strain of the species, and is designated BL-540 (ATCC 33623). L. jordanis had a partial relationship to L. bozemanii by direct fluorescent-antibody tests but was unrelated to L. pneumophila, L. dumoffii, L. micdadei, L. gormanii, or L. longbeachae. Legionella phenotypic characteristics, including large amounts of branched-chain cellular fatty acids, were shown by the isolates. Studies of DNA relatedness showed that the two cultures of L. jordanis were only slightly related to the six previously described species of Legionella but were more than 90% related to each other. Indirect fluorescent-antibody tests with human sera suggested that unrecognized human infections with L. jordanis may be occurring.
GM1 ganglioside enzyme-linked immunosorbent assay for detection of heat-labile enterotoxin produced by human and porcine Escherichia coli strains
Authors: Gustafsson B , Möllby R
Published: January 2020

Human and porcine enterotoxigenic strains of Escherichia coli were cultivated in tryptone-yeast extract medium or brain heart infusion broth and tested for production of heat-labile enterotoxin by the GM1 ganglioside enzyme-linked immunosorbent assay (GM1-ELISA) and the Y1 adrenal cell assay. When testing for enterotoxigenicity by the GM1-ELISA technique, homologous antisera for human and porcine heat-labile enterotoxins had to be used to detect enterotoxigenic strains of human and porcine origin, respectively. This observation indicates a serological difference between the heat-labile enterotoxins produced by human and porcine strains. Furthermore, brain heart infusion broth was found to have an inhibitory effect on detection of enterotoxin both in the GM1-ELISA and in a toxin-binding modification of the Y1 adrenal cell test, but not in the conventional adrenal cell assay.
Detection of Japanese encephalitis virus immunoglobulin M antibodies in serum by antibody capture radioimmunoassay
Authors: Burke D S , Nisalak A
Published: January 2020

An assay for detecting human immunoglobulin M (IgM) antibodies to Japanese encephalitis (JE) virus was developed by using the antibody capture solid-phase radioimmunoassay approach (JE IgM ACRIAAA). Heavy-chain-specific goat antihuman IgM was first bound to the wells of a polyvinyl microtiter plate, and successive steps involved sequential binding of test sample IgM, acetone-extracted mouse brain JE antigen, and (125)I-labeled flavivirus hyperimmune human IgG. Among 20 patients hospitalized in Bangkok with clinical diagnoses of acute encephalitis, and with acute flavivirus infections proven by hemagglutination inhibition (HAI) serology, 16 had detectable (positive/negative [P/N] ratio, greater than 3.0) JE IgM ACRIA antibodies in the acute-phase serum specimen, and 19 had such antibodies in the convalescent-phase serum specimen. Convalescent patient sera regularly had higher P/N values than the corresponding acute-phase sera (mean +/- 1 standard deviation = 13.0 +/- 9.3 with acute-phase sera and 25.8 +/- 19.6 with convalescent-phase sera). JE virus-infected patients with HAI serological responses indicative of a primary flavivirus infection had higher JE IgM ACRI P/N responses than did those patients whose serological response indicated past exposure to other flaviviruses. None of 70 serum specimens from healthy Thai adults and children with serum JE HAI antibodies had detectable JE IgM ACRIA activity (P/N ratios all less than or equal to 3.0). Biological false-positives with low P/N ratios (range, 3 to 15) were found in sera from patients with acute or recent infections with flaviviruses other than JE virus but could be differentiated by the fact that these sera gave higher P/N ratios with homologous antigens than with JE virus. False-positive reactions with low P/N ratios (range, 3 to 6) due to serum rheumatoid factor activity were differentiated by testing with control antigen. The JE IgM ACRIA technique permits a rapid, accurate diagnosis of acute JE virus infections in both patients with and those without previous exposure to other flaviviruses.
Detection of antibodies to pneumococcal capsular polysaccharides by enzyme-linked immunosorbent assay
Authors: Pedersen F K , Henrichsen J
Published: January 2020

An enzyme-linked immunosorbent assay for the detection of immunoglobulin G, M, and A antibodies against each of the 14 polysaccharide antigens contained in a contemporary pneumococcal vaccine is described. A mean total antibody fold increase above 2 from prevaccination to 4 weeks postvaccination serum samples was found for all antigens in 12 healthy adults and 6 children. Fifty-five percent of all single fold increases determined were above 2 in the adults; the value was 64% in children. Type-specific polysaccharide antibody of the immunoglobulin G class was predominant in 4 weeks postvaccination serum samples. Further studies with assays such as the one described may lead to a better understanding of the immune response to pneumococcal polysaccharides in both normal subjects and patients at increased risk of pneumococcal infection.
Screening of Urine Cultures by Three Automated Systems
Authors: Pezzlo Marie T. , Tan Grace L. , Peterson Ellena M. , De La Maza Luis M.
Published: January 2020

A study was conducted to compare three automated systems and the Gram stain for their ability to detect significant bacteriuria. A total of 1,000 urine specimens were evaluated by Autobac MTS (General Diagnostics), Auto Microbic system (AMS; Vitek Systems, Inc.), and MS-2 (Abbott Laboratories) and compared with a semiquantitative culture plate method. Two hundred thirty-nine (23.9%) specimens had colony counts of >10
5
colony-forming units (CFU)/ml by the culture plate method (group I). Of these, 204 (85.3%) were positive by Autobac, 198 (82.8%) were positive by AMS, and 179 (74.9%) were positive by MS-2. When pure cultures of diphtheroids, lactobacilli, and viridans streptococci not group D were considered contaminants and therefore excluded, there were 118 specimens containing pure cultures of probable pathogens. The percentage of significant isolates detected was 97.4% (115 of 118) by the Gram stain, 96.6% (114 of 118) by Autobac, and 95.8% (113 of 118) by AMS and MS-2. The average detection time for all organisms was 2.2 h by Autobac, 6.1 h by AMS, and 1.8 h by MS-2; therefore, all three methods were more rapid than the 18- to 24-h standard plate culture method. One hundred sixty-one (16.1%) specimens had colony counts of 10
4
to 10
5
CFU/ml (group II). The probable pathogens not detected in this group were two (1.2%) by Autobac and MS-2 and three by AMS (1.9%). The average detection time for group II was 4.2 h by Autobac, 8.9 h by AMS, and 3.8 h by MS-2. Six hundred specimens had colony counts of <10
4
CFU/ml. Of these, 188 had colony counts equal to 10
3
and <10
4
CFU/ml (group III), and 412 cultures were below detectable limits by the standard plate method (group IV). Less than 37 and 15% of groups III and IV, respectively, were detected by instrumentation. Average detection times for groups III and IV were 4.6 and 4.8 h by Autobac, 10 and 11 h by AMS, and 4.2 and 4.4 h by MS-2. The cost of supplies and technical time with Gram stain, Autobac, and MS-2, when used as screening methods, were comparable and considerably less expensive than for the reference method. The AMS was the least expensive system when the cost for identifying probable pathogens was included.
Routine test for in vitro differentiation of pathogenic and apathogenic Listeria monocytogenes strains
Authors: Skalka B , Smola J , Elischerová K
Published: January 2020

The exosubstance of Rhodococcus equi in a prepurified form strongly enhanced the hemolytic effect of certain strains of Listeria monocytogenes. The strains which produced positive synergic hemolysis with this exosubstance were also pathogenic for guinea pigs and white mice. The other strains, which remained nonhemolytic in the presence of the R. equi. exosubstance, were apathogenic for those animals. A routine test was devised for the in vitro determination of the pathogenicity of L. monocytogenes strains.
Rapid isolation of Yersinia spp. from feces
Authors: Weissfeld A S , Sonnenwirth A C
Published: January 2020

Direct plating or cold enrichment or both have been used to isolate Yersinia spp. from feces. Freeze-shock double enrichment and KOH treatment have been recommended for recovery of Yersinia enterocolitica from surface waters and food, respectively. These techniques were evaluated as alternatives for rapid recovery of Yersinia spp. from feces. Stool samples were homogenized in buffered saline and autoclaved. Escherichia coli. Klebsiella pneumoniae, and Pseudomonas aeruginosa were each added to the suspension at a final concentration of 1.5 x 10(6) colony-forming units per ml. Yersinia cells were then added to a final concentration of 1.5 x 10(3), 1.5 x 10(4), 1.5 x 10(5), or 1.5 x 10(6) colony-forming units per ml. A total of 21 strains of Y. enterocolitica, 2 of Yersinia kristensenii, and 1 each of Yersinia intermedia and Yersinia fredriksenii were tested. For freeze-shock double enrichment, seeded stool samples were frozen overnight (-70 degrees C), transferred successively to m-tetrathionate broth (6 h. 37 degrees C) and selenite broth (2 h 37 degrees C), and plated on MacConkey, salmonella-shigella, and cellobiose-arginine-lysine agars for quantitation. For KOH treatment, seeded stool samples were mixed with 0.5% KOH at a ratio of 1:2 for 2 min and plated as described above. E. coli, K. pneumoniae, and P. aeruginosa were virtually eliminated after either method was used. All Yersinia strains were recovered after KOH treatment even at the lowest initial concentration (1.5 x 10(3) colony-forming units per ml). However, after freeze-shock double enrichment, not all strains were retrievable, and those isolates which were recovered were grown only from samples containing the highest number of Yersinia strains (1.5 x 10(6) colony-forming units per ml). KOH treatment of stool samples seems to be a viable substitute for more protracted methods of recovering Yersinia spp.
Characterization of atypical Candida tropicalis and other uncommon clinical yeast isolates
Authors: Schlitzer R L , Ahearn D G
Published: January 2020

Clinical yeast isolates representing alpha-glucoside-deficient variants of Candida tropicalis, C. lusitaniae, atypical C. albicans, and Saccharomyces cerevisiae were characterized. Additional physiological tests, including cellobiose fermentation, rhamnose assimilation, and triphenyl tetrazolium chloride reduction, are recommended for the detection and presumptive identification of uncommon Candida spp. in the clinical laboratory.
Solid-phase immune electron microscopy-double-antibody technique for rapid detection of papovaviruses
Authors: Giraldo G , Beth E , Lee J , de Harven E , Chernesky M
Published: January 2020

The solid-phase immune electron microscopy-double-antibody technique, which takes less than 1 h to perform, was applied as a rapid, sensitive, and specific diagnostic tool in the demonstration of papovavirus particles. BK virus propagated in 82C human skin fibroblasts and a monospecific high-titer immune serum to BK virus were used to establish the test procedure. When Formvar-carbon-coated grids were treated with appropriately diluted antibody, a 28-fold increase of virus particles per square micrometer was observed. Viewing of the virus particles was facilitated by the addition of a second "decorator" antibody. BK virus preparations at concentrations of 10(2) to 10(3) PFU/ml could be detected by this technique. There was no cross-reaction with mouse polyomavirus.
Urine culture transport tubes: effect of sample volume on bacterial toxicity of the preservative
Authors: Nickander K K , Shanholtzer C J , Peterson L R
Published: January 2020

Stable bacterial counts in urine specimens before culture are necessary to assure the accurate diagnosis of urinary tract infections. Preservative-containing tubes are commercially available for urine transport. As these tube containers are not always filled to the manufacturer's specifications, we studied the effects of stabilizer formation with low urine volumes. The Sage Urine Culture Tube and the Becton-Dickinson Urine Culture Kit were evaluated by using 30 cultures diluted in urine to 10(5) colony-forming units per ml. Both tube types were injected with 1, 2, 3, and 4 to 5 ml (tube capacity) of urine containing each culture. Specimens were held at 22 degrees C and cultured at 0, 4, and 24 h. Colony counts were corrected for the dilution due to the preservative. The Becton-Dickinson Urine Culture Kits were toxic to Escherichia coli and Klebsiella pneumoniae in specimens containing up to 2 ml of urine, and the minimum usable amount of urine for reliable results was 3 ml. The Sage Urine Culture Tube maintained the number of bacteria in 1 to 4.5 ml of urine in 83% of the specimens. However, the Sage tube was toxic to E. coli when held for 24 h. Quantitative counts of enterococci tended to significantly increase in specimens that contained 2 ml or more of urine with either system. The limitation of preservative-containing tubes for urine transport need to be recognized in order to avoid false-positive and false-negative results.
Increased bactericidal activity of dilute preparations of povidone-iodine solutions
Authors: Berkelman R L , Holland B W , Anderson R L
Published: January 2020

Recent confirmation of intrinsic bacterial contamination of 10% povidone-iodine solution has raised questions regarding the bactericidal mechanism of iodophors and the possibility for survival of vegetative bacterial cells in iodophor solutions. In this laboratory investigation, five different species were exposed to various dilutions of three commercial preparations of 10% povidone-iodine solution; survival was assessed after exposure for time periods varying between 0 and 8 min. All brands of povidone-iodine solution tested demonstrated more rapid killing of Staphylococcus aureus and Mycobacterium chelonei at dilutions of 1:2, 1:4, 1:10, 1:50, and 1:100 than did the stock solutions, S. aureus survived a 2-min exposure to full-strength povidone-iodine solution but did not survive a 15-s exposure to a 1:100 dilution of the iodophor. Both stock and dilute preparations of 10% povidone-iodine solution demonstrated rapid bactericidal action against Klebsiella pneumoniae, Pseudomonas cepacia, and Streptococcus mitis.
Serological and genotypic diversity among serogroup 5- reacting environmental Legionella isolates
Authors: Garrity G M , Elder E M , Davis B , Vickers R M , Brown A
Published: January 2020

Five strains of bacteria (strains 684, 687, U7W, U8W, and MICU-B) that were biochemically and morphologically indistinguishable from Legionella pneumophila were recovered from the environment. Strains U7W, U8W, and MICU-B were antigenically identical to L. pneumophila strain Dallas 1E (serogroup 5), as determined by direct fluorescent antibody staining and immunodiffusion. Although strains 694 and 687 shared antigenic determinants with L. pneumophila Dallas 1E, they could be distinguished by immunodiffusion and differential absorption studies. However, as determined by DNA hybridization, the antigenically distinct strains 684 belong to the same DNA homology cluster as previously described authentic strains of L. pneumophila, whereas strain U7W, U8W, and MICU-B belong to a separate homology group. Therefore, two groups could be identified among these environmental isolates; one represents an antigenic subclass of serogroup 5 L. pneumophila (strains 684, and 687), and the other, although antigenically indistinguishable from serogroup 5 L. pneumophila, probably represents a new Legionella species (strains U7W, U8W, and MICU-B).
Safety considerations in handling exoantigen extracts from pathogenic fungi
Authors: Standard P G , Kaufman L
Published: January 2020

Currently, exoantigen test procedures for identifying mycelial form cultures of pathogenic molds require that the fungi being extracted be treated with thimerosal to render them safe for handling. Recent studies have demonstrated that thimerosal may not be fungicidal. In view of these reports, we investigated the effects of thimerosal and formaldehyde on a variety of exoantigen preparations. Mature mycelial form fungal cultures, including cultures of Blastomyces dermatitidis, Coccidioides immitis, and Histoplasma capsulatum and morphologically similar fungi, were grown on Sabouraud dextrose agar slants and treated with 0.02, 0.04, and 0.08% thimerosal for 24 and 48 h and with 0.2 and 0.5% formaldehyde for 24 and 48 h. We found that 0.5% formaldehyde killed all of the fungi studied, whereas 0.2% formaldehyde permitted the growth of only one fungus; 0.02, 0.04, and 0.08% thimerosal were fungistatic. Furthermore, 0.2 and 0.5% formaldehyde and 0.08% thimerosal affected certain antigens adversely. For those investigators who prefer to use 0.02% thimerosal and to work with sterile extracts, we recommend that the procedure be modified, and we advocate sterilization of extracts by passage through membrane filters.
Passive hemagglutination test for detection of antibody to gonococcal ribosomal antigen in sera from patients with asymptomatic gonorrhea
Authors: Kita E , Kashiba S
Published: January 2020

Ribosomal fractions were obtained from a culture of type 2 Neisseria gonorrhoeae strain P-17 which was isolated from a patient with an acute gonococcal infection; these fractions were purified to eliminate the components of the outer membrane complex by affinity chromatography (Sepharose-anti-outer membrane complex antibody conjugates were used as the solid immunosorbent), and the resulting preparation was designated the purified ribosomal fraction, The purified ribosomal fraction was used to detect antibody activity in sera obtained from culture-positive asymptomatic carriers and healthy controls by a passive hemagglutination test. This passive hemagglutination test had a specificity of 100% for both sexes and sensitivities of 99.4 and 88.2% for female and male carriers, respectively, when an antibody titer of more than 1:3 was defined as abnormal. Absorption of the sera with nongonococcal organisms did not affect the antibody activity, and no significant difference in antigenicity among various N. gonorrhoeae strains was observed in ribosomal fractions. An enzyme-linked immunosorbent assay was also used to measure the relative amounts of specific antibodies to the purified ribosomal fraction, and this assay revealed that the anti-purified ribosomal fraction antibodies were immunoglobulin G.
Atypical biogroups of Escherichia coli found in clinical specimens and description of Escherichia hermannii sp. nov
Authors: Brenner D J , Davis B R , Steigerwalt A G , Riddle C F , McWhorter A C , Allen S D , Farmer J J , Saitoh Y , Fanning G R
Published: January 2020

DNA relatedness was used to define the biochemical boundaries of Escherichia coli. A large number of biochemically atypical strains were shown to belong to biogroups of E. coli. These included strains negative in reactions for indole, all three decarboxylases, D-mannitol, lactose, or methyl red and strains positive in reactions for H2S, urea, citrate, KCN, adonitol, myo-inositol, or phenylalanine deaminase. Frequency and source data are presented for these atypical E. coli biogroups. One group of KCN-positive, cellobiose-positive, yellow-pigmented strains was 84 to 91% interrelated but only 35 to 45% related to E. coli. The name Escherichia hermannii sp. nov. is proposed for this group of organisms that was formerly called Enteric Group 11 by the Enteric Section, Centers for Disease Control, Atlanta, GA. Twenty-nine strains of E. hermannii have been isolated in the United States from a variety of clinical sources, principally wounds, sputum, and stools. Three additional strains were isolated from food. E. hermannii strains are gram-negative, oxidase-negative, fermentative, motile rods. In addition to yellow pigment and positive KCN and cellobiose tests, the biochemical reactions characteristic of 32 strains of E. hermannii were as follows: gas from D-glucose, acid from D-glucose, maltose, D-xylose, L-arabinose, L-rhamnose, and D-mannitol; no acid from adonitol or inositol; variable acid production from lactose and sucrose; positive tests for indole, methyl red, and mucate; negative tests for Voges-Proskauer. Simmons citrate, H2S, urea, phenylalanine deaminase, and gelatin hydrolysis; negative or delayed test for L-lysine decarboxylase and negative test for L-arginine dihydrolase; and positive test for ornithine decarboxylase. E. hermannii strains were resistant to penicillin, ampicillin, and carbenicillin and sensitive to other commonly used antibiotics. Wounds account for almost 50% of human isolates of E. hermannii, followed by sputum or lung isolates (ca. 25%) and stool isolates (20%).