bluuebunny/crossref_metadata_2025 huggingface dataset:
Performance of the prompt system in identification and antimicrobial susceptibility testing of clinical isolates
Authors: Flejzor B , Bokkenheuser V D
Published: January 2020

The rapid 3M Prompt inoculation system was compared with the traditional log-phase system for Autoscan identification and antibiotic susceptibility testing (MIC) of 188 recent clinical isolates. The two systems were equally effective for gram-negative rods; the Prompt system was slightly superior for the determination of MICs for gram-positive organisms.
Recurrent intraabdominal abscess caused by Salmonella paratyphi C
Authors: Freidin N , Merzbach D , Kleinhaus U , Eidelman S
Published: January 2020

We describe a patient who suffered from recurrent intraabdominal abscesses. The last two of the three abscesses were certainly caused by Salmonella paratyphi C. The time interval between the first and the second abscess was 25 years, and that between the second and the third abscess was 20 years. Single infection with this microorganism is very rare in Israel, with only four known cases in the last 20 years. The annual frequency in the United States is 0 to 2 cases per year. In recent years, this infection has also been very rare in other parts of the world. Our case is unique as it recurred two or three times. The infection was probably dormant for a very long time. During the dormant years, the patient was clinically healthy. To our knowledge a recurrent infection with this microorganism has not previously been reported in the literature.
Monoclonal antibodies against respiratory syncytial virus and their use for rapid detection of virus in nasopharyngeal secretions
Authors: Pothier P , Nicolas J C , de Saint Maur G P , Ghim S , Kazmierczak A , Bricout F
Published: January 2020

We developed five monoclonal antibodies against respiratory syncytial virus. Three of these (23A3, 12A4, and 18B2) were used in an indirect fluorescent antibody test, and the results were compared with those of a similar indirect fluorescent test with commercial anti-respiratory syncytial virus serum. The results obtained with antibody 18B2 and commercial anti-respiratory syncytial virus serum were identical, whereas with antibodies 23A3 and 12A4 the incidence of positive identifications was around 50%.
Collaborative evaluation of the UniScept qualitative antimicrobial susceptibility test
Authors: D'Amato R F , Isenberg H D , McKinley G A , Hochstein L , Cleri D J
Published: January 2020

The UniScept system (Analytab Products, Plainview, N.Y.) is a commercially prepared microdilution antimicrobial susceptibility test for the determination of qualitative susceptibility results for gram-negative and gram-positive bacteria. The system showed excellent correlation with the reference agar diffusion approach for organisms from clinical specimens and with stock and reference cultures. Intra- and interlaboratory reproducibility was high.
Serogrouping of Clostridium difficile strains by slide agglutination
Authors: Delmee M , Homel M , Wauters G
Published: January 2020

Six different agglutinating antisera were obtained by immunizing rabbits with Formol-treated strains of Clostridium difficile. After appropriate absorption, these antisera were used to define six serogroups designated by the letters A, B, C, D, F, and G. Altogether, 315 strains of C. difficile from various origins were tested for slide agglutination by these antisera; 312 (99%) of them were agglutinated by one of these antisera. A and C were the most common serogroups. An excellent correlation, ranging from 85 to 100%, was found between the serogroup and the toxigenicity of the strains. The correlation between serogroup and sorbitol fermentation was higher, ranging from 89 to 100%. The results of this typing were compared with the clinical origin of the strains. Only strains of serogroups A, C, and D were isolated in 153 cases of antibiotic-associated diarrhea. This series included strains from three outbreaks; all the strains in two of the outbreaks belonged to serogroup C, and in the third, all the strains belonged to serogroup A. Strains of serogroups B, F, and G were only found in the stools of asymptomatic neonates or young children. In the latter samples, strains of serogroups A and D were found in the same ratio as in adults with antibiotic-associated diarrhea, but strains of serogroup C were seldom isolated. In patients treated with antineoplastic drugs and suffering from diarrhea, the distribution of the strains was the same as in cases of antibiotic-associated diarrhea.
Proposed quality control and interpretive criteria for disk diffusion susceptibility testing with enoxacin
Authors: Rudrik J T , Cavalieri S J , Britt E M
Published: January 2020

The standardized disk diffusion test, in which a 10-micrograms enoxacin disk is used, was performed and microbroth dilution MICs were determined to establish individual test control values with Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923, and S. aureus ATCC 29213. In addition, regression analysis correlating inhibitory zone diameter with MICs for approximately 400 gram-negative clinical isolates was performed. Based on linear regression and error rate-bounded analyses, criteria for the category calls of isolates are proposed.
Microcarriers in combination with enzyme immunofiltration and immunofluroescence for the detection of herpes simplex virus antigens in culture
Authors: Rossier E , Scalia V , Phipps P H , Kennedy D A , Brodeur B
Published: January 2020

A microcarrier culture system in combination with enzyme immunofiltration with a herpes simplex virus (HSV) group monoclonal antibody was found to be as sensitive as immunofluorescence for the detection of HSV type 1 (HSV-1) in cell cultures and to give specific identification at the same time as the appearance of the cytopathic effect with very high infectious inocula and within 10 to 24 h of the appearance of the cytopathic effect with very high infectious inocula and within 10 to 24 h of the appearance of the cytopathic effect with low to high inocula. Multiplicities of infection from 10(-5) to 10(1) were tested at 8 to 96 h postinfection. When applied to the identification of HSV-1 and HSV-2 in cultures of clinical samples, the system detected HSV antigens in 50% of the samples after 2 days and in 100% of the samples after 3 days. With 2 ml of microcarrier suspension and with 50 to 300 microliter per sample, several portions are available for replicate and sequential sampling without destroying the culture. The system requires only that 2 X 10(5) microcarriers be added to the culture tube at the time or before it is seeded with cells, at an extra cost of 6 cents (U.S.) per tube and little extra labor.
Comparative evaluation of the enzyme-linked immunosorbent assay in the laboratory diagnosis of brucellosis
Authors: De Klerk E , Anderson R
Published: January 2020

An enzyme-linked immunosorbent assay was adapted to measure total and Brucella abortus-specific immunoglobulin M antibodies. The results were compared with those of conventional serological tests for B. abortus antibody on the sera of a number of normal controls, apparently healthy occupationally exposed workers, and patients with suspected acute brucellosis. Relative to other tests, the B. abortus enzyme-linked immunosorbent assay was found to be both highly sensitive and highly specific. The serological results obtained in occupationally exposed workers indicate a higher "normal range" for this group and therefore a possibility of false-positive results and overdiagnosis. It is therefore important to establish a separate "normal range" for occupationally exposed workers. Investigation of patients with acute brucellosis showed that the enzyme-linked immunosorbent assay for immunoglobulin M was the most sensitive serodiagnostic test and was likely to be of value in the serodiagnosis of acute brucellosis in occupationally exposed workers.
Modification of a direct enzyme-linked immunosorbent assay for the detection of immunoglobulin G and M antibodies to pneumococcal capsular polysaccharide
Authors: Messina J P , Hickox P G , Lepow M L , Pollara B , Venezia R A
Published: January 2020

In contrast to the usual indirect enzyme-linked immunosorbent assay (ELISA) method for detection of antibody responses, a modified direct ELISA technique was used to measure immunoglobulin G (IgG) and IgM responses to pneumococcal capsular types 1, 3, 9N, and 23F in humans. Individual capsular polysaccharides were covalently bound to poly-L-lysine before adsorption to the solid phase. The coupling reaction was enhanced by maintenance of a constant pH of 8.2 after the addition of all reactants. The evaluation of four diluents (phosphate-buffered saline [PBS]-Tween; PBS-Tween plus 10% fetal calf serum; PBS-Tween plus 10% bovine serum albumin; and PBS-Tween plus 20% normal goat serum) showed that the sensitivity and specificity of the assay was increased with normal goat serum (10-fold). Serum samples from 10 subjects immunized with polyvalent pneumococcal vaccine were tested by direct ELISA and by radioimmunoassay. At 4 weeks postimmunization, the ELISA method showed that IgG was the predominant antibody and that IgM responses were lower or had diminished. Isotype shifts during this period would have been undetected by the radioimmunoassay method. The changes in antibody response measured by ELISA were comparable to the radioimmunoassay results. The direct ELISA method for the detection of antipneumococcal capsular antibody has been found to be a sensitive and reproducible assay for the detection of IgG and IgM antibodies.
Evaluation of an enzyme-linked immunosorbent assay for treponemal antibody
Authors: Stevens R W , Schmitt M E
Published: January 2020

A new enzyme-linked immunosorbent assay with Treponema pallidum antigen bound to ferrous metal beads (Syphilis Bio-EnzaBead; Litton Bionetics Laboratory Products) was compared with the standard fluorescent treponemal antibody-absorption test for syphilis. Bio-EnzaBead and fluorescent treponemal antibody-absorption tests were done on 218 specimens from documented cases of syphilis, on 315 sera from individuals with diseases other than syphilis, and on sera submitted to a public health laboratory for premarital (304 specimens) or diagnostic (501 specimens) tests for syphilis. Agreement between the Bio-EnzaBead and reference tests ranged from 93.0% for sera for the diagnostic test to 99.5% for sera from patients with syphilis. The overall agreement among the 1,338 sera tested was 96.3%. The reproducibility of the Bio-EnzaBead test with 60 coded sera of graded reactivity was 97%. The test is easy to perform, the indicator results are clear and unequivocal, and the findings are comparable to those of the fluorescent treponemal antibody-absorption test.
Feasibility study of disk diffusion susceptibility tests with Mueller-Hinton broth solidified with Gelrite, an agar substitute
Authors: Barry A L , Creitz L , Packer R R
Published: January 2020

Feasibility studies were done to determine whether a new agar substitute, Gelrite gellan gum, could be used to prepare a solid Mueller-Hinton medium for disk diffusion susceptibility tests. Mueller-Hinton broth was combined with 0.43% of the gellan gum and 0.75% KCI. The resulting medium had performance characteristics similar to those of Mueller-Hinton agars; however, zones on the gellan gum media tended to be a little larger. Significant differences among Mueller-Hinton broths and among Mueller-Hinton agars from different manufacturers were documented: zones on different lots of the gellan gum were more consistent. The Mueller-Hinton broth-gellan gum medium appears to represent a satisfactory alternative to agar media. However, because somewhat larger zones were seen on the gellan gum plates, further study will be needed to develop quality control limits and interpretive zone size standards for tests on this new medium.
Serotypic characterization of rotaviruses derived from asymptomatic human neonatal infections
Authors: Hoshino Y , Wyatt R G , Flores J , Midthun K , Kapikian A Z
Published: January 2020

Nineteen rotavirus strains derived from asymptomatic neonates (seven from England, five from Australia, two from Venezuela, and five from Sweden) were successfully cultivated in primary African green monkey kidney cell cultures, serotyped by plaque reduction neutralization tests, subgrouped by indirect enzyme-linked immunosorbent assay, and electropherotyped by polyacrylamide gel electrophoresis. All 19 strains were shown to fall into one of the four known human serotypes; serotype 1 (all Venezuelan strains), serotype 2 (all Swedish strains), serotype 3 (all Australian strains), or serotype 4 (all English strains). Hyperimmune guinea pig serum raised against the Venezuelan strain (M37) neutralized not only serotype 1 (strain Wa) but also serotype 4 (strain St. Thomas no. 3) viruses to a similar degree. The English, Australian, and Venezuelan isolates were found to belong to subgroup 2, and the Swedish strains were subgroup 1 viruses. The potential importance of these rotaviruses obtained from neonates as possible vaccine candidates is discussed.
DNA probe specific for Legionella pneumophila
Authors: Grimont P A , Grimont F , Desplaces N , Tchen P
Published: January 2020

A procedure for preparing a DNA probe to be used in the specific detection of Legionella pneumophila by dot or colony hybridization has been devised. When total DNA from L. pneumophila was used as a radioactive probe, cross-hybridization occurred with DNA from many other species belonging to various families (including Legionellaceae, Enterobacteriaceae, Pseudomonadaceae, and Vibrionaceae). Cross-hybridizing restriction fragments in L. pneumophila ATCC 33152 DNA were identified on Southern blots. When unlabeled DNA from strain ATCC 33152 was cleaved by endonuclease BamHI, the DNA fragments cross-hybridizing with the labeled DNA from all of the other species and genera tested (or with Escherichia coli 16 + 23 S RNA) had a size of 21.4 and 16.2 kilobase pairs (major bands) and 28.0, 12.8, and 10.1 kilobase pairs (minor bands). BamHI restriction fragments of L. pneumophila DNA deprived of the cross-hybridizing fragments were pooled and used as a probe for the detection of L. pneumophila. This probe proved to be specific for L. pneumophila in colony and dot hybridization. It can potentially be used for the detection of L. pneumophila in clinical and water samples. The procedure described can be readily applied to the preparation of probes specific for phylogenetically isolated bacterial species other than L. pneumophila.
Evaluation of the Micro-ID system for the identification of Yersinia pestis
Authors: Harrison D N , Williams J E
Published: January 2020

One hundred isolates of Yersinia pestis identified by conventional means were tested by the Micro-ID system to assess its reliability for distinguishing Y. pestis from other members of the family Enterobacteriaceae. The Micro-ID system gave Y. pestis as a choice for the identification of 89 of these cultures, although not always as the first choice. Most nitrate-negative strains of Y. pestis keyed out with Yersinia pseudotuberculosis as first choice and Y. pestis as second or fourth choice.
Infections acquired in clinical laboratories in Utah
Authors: Jacobson J T , Orlob R B , Clayton J L
Published: January 2020

We reviewed laboratory-acquired infections occurring in Utah from 1978 through 1982. Written and telephone interviews of supervisors of 1,191 laboratorians revealed an estimated annual incidence of 3 laboratory-acquired infections per 1,000 employees. Infections, in order of frequency, included hepatitis B (clinical cases), shigellosis, pharyngitis, cellulitis, tuberculosis (skin test conversion), conjunctivitis, and non-A, non-B hepatitis. One-half of large laboratories (over 25 employees), but only 12% of smaller laboratories, reported infections. The annual incidence, however, at smaller laboratories was more than three times greater than at large laboratories (5.0 versus 1.5 per 1,000; P less than 0.05, chi-square test). Microbiologists were at greatest risk of infection, with an incidence of almost 1%, followed by generalists and phlebotomists. Shigellosis was acquired only by microbiologists and accounted for more than half of their infections. The most common laboratory-acquired infection, hepatitis B, affected a microbiologist, a hematologist, a phlebotomist, a pulmonary blood gas technician, and a blood bank technologist who died from her illness. Clinical cases of hepatitis B occurred at a rate 10 times higher than the rate in the general U.S. population. The incidence of tuberculosis skin test conversion was intermediate between rates reported for hospital employees and for the state of Utah.
Enzyme-linked immunosorbent assay for determination of antibodies against herpes simplex virus types 1 and 2 in human sera
Authors: Hampar B , Zweig M , Showalter S D , Bladen S V , Riggs C W
Published: January 2020

A rapid and reproducible enzyme-linked immunosorbent assay (ELISA) is described for determining antibodies in human sera against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). The sera were absorbed for 30 min with heterologous virus-infected-cell extracts to remove cross-reacting antibodies and then were applied to ELISA plates containing the target antigens, immunoaffinity-purified HSV-1 glycoproteins gC and gD and HSV-2 glycoproteins gD and gF. The absorbance index, defined as the ratio of A414 generated by a serum sample absorbed with a heterologous virus-infected-cell extract versus the A414 of a serum sample absorbed with an uninfected-cell extract, was used to determine the presence or absence of antibodies to HSV-1 and HSV-2. Results of the ELISA for detecting antibodies against HSV-2, when compared with results obtained for the same sera by the microneutralization test, showed an index of overall agreement of 91%. Results of the ELISA for detecting antibodies against HSV-1, when compared with microneutralization test results for sera negative for HSV-2 antibodies but positive for HSV antibodies by ELISA, showed an index of agreement of 99%.
Rapid, colorimetric method for the detection of microorganisms in blood culture
Authors: Wallis C , Melnick J L
Published: January 2020

A semiautomated instrument (Septi-Screen) and method are described that semiquantify bacteria colorimetrically in blood-inoculated cultures in 2 to 8 h. If the blood culture is positive, concentrates (of viable bacteria) are obtained free of blood cells. In pediatric cases and in septicemia attributed to indwelling catheters where bacteremia is often manifested by high bacterial counts, the total time required to obtain bacterial concentrates is only 2 to 3 h. In brief, at various intervals after inoculation of 10 ml of blood into 100 ml of culture fluid, 25-ml blood culture samples are passed through the Septi-Filter. Erythrocytes and leukocytes pass into the filtrate and are discarded, but bacteria are electrostatically retained on the filter surface. The entrapped bacteria are backflushed and recovered as a concentrate that is processed through a second filter (Septi-Count) for colorimetric quantitation. The bacteria are stained, and the filter fibers are decolorized. The color intensity of the filter surface is proportional to the number of bacteria in the concentrate processed. The total Septi-Screen processing time is 3 to 5 min.
Standardization of a hyphal inoculum of aspergilli for amphotericin B susceptibility testing
Authors: Bezjak V
Published: January 2020

Standardized, evenly dispersed hyphal suspensions served as the inoculum in a microtiter technique for amphotericin B antifungal susceptibility testing. Preliminary testing with six strains of Aspergillus fumigatus and A. flavus produced consistent and reproducible results at 30 degrees C over 24 h. The observed amphotericin B MICs required for hyphae (0.3 to 0.6 microgram/ml) were comparable to MICs required for conidia (0.16 to 0.6 microgram/ml). The results were evaluated and compared with previously published information.
Antibody response to cytomegalovirus polypeptides captured by monoclonal antibodies on the solid phase in enzyme immunoassays
Authors: Cremer N E , Cossen C K , Shell G R , Pereira L
Published: January 2020

Antibodies to different cytomegalovirus (CMV) polypeptide antigens, captured by monoclonal antibodies coated on the solid phase of an enzyme immunoassay test, were analyzed in 42 serum pairs submitted for serodiagnosis of CMV infection. Three CMV antigens, captured on the solid phase by three monoclonal antibodies of different specificities, designated CH92-1, CH65-1, and CH16-1, were glycoproteins A (gA), gC, and gD, respectively; and one antigen, captured by CH23, was a polypeptide with an apparent molecular weight of 150,000, possibly associated with the nucleocapsid. Of these four CMV antigens, gA captured by CH92-1 was most effective in eliciting an antibody response. Antibody to this antigen was present in serum samples at a higher concentration in primary and reactivated infection and persisted longer than did antibody to the other tested antigens. In contrast, antibody to antigen captured by CH23 was at a lower concentration, rose more slowly in infection, and persisted for a shorter time than did antibody to the other antigens. Antibody response to gC and gD was intermediate in concentration and temporal appearance compared with the antibody response to gA and to the polypeptide bound by CH23. An enzyme immunoassay on paired serum samples with the captured glycoproteins as antigen was equal for the detection of current infection to an enzyme immunoassay with the whole CMV antigen from infected cell lysates. Enzyme immunoassays with either the CMV glycoproteins or the whole CMV antigen from infected cell lysates were superior to a complement fixation test with a glycine extract antigen for serodiagnosis of current infection.
Rapid species identification of group C streptococci isolated from horses
Authors: Bannister M F , Benson C E , Sweeney C R
Published: January 2020

Two commercial systems, the API 20S (Analytab Products, Plainview, N.Y.) and the Rapid Strep (API System S.A., Montalieu-Vercieu, France), were evaluated for ease of use and accuracy in the rapid identification of group C streptococci isolated from horses. A total of 85 Streptococcus isolates were tested, including S. equi (67 isolates), S. zooepidemicus (13 isolates), and S. equisimilis (5 isolates). All S. equi and S. zooepidemicus isolates were correctly identified within 24 h by the Rapid Strep system. Specific grouping sera was necessary to distinguish between S. equisimilis and group G or L strains. The API 20S system did not provide species identification of any of these isolates. An identification of randomly selected isolates to species level was performed by conventional methods and confirmed the identification derived through the Rapid Strep system. Our results indicate that the Rapid Strep system is a valuable aid for species identification of equine isolates of group C streptococci.
DNA restriction endonuclease analysis of Mycobacterium bovis and other members of the tuberculosis complex
Authors: Collins D M , De Lisle G W
Published: January 2020

DNA preparations from 24 New Zealand isolates, two reference strains of Mycobacterium bovis, and one reference strain each of Mycobacterium microti, Mycobacterium africanum, and Mycobacterium tuberculosis were characterized by restriction endonuclease analysis. Twenty-five restriction enzymes were investigated. The clearest differences in M. bovis patterns were obtained with the enzymes BstEII and BclI. These produced four and five different patterns, respectively, for the 24 local isolates. When the results from both enzymes were considered, seven different combinations were obtained. The patterns produced for the two reference strains of M. bovis could be distinguished from each other and also from the patterns produced for the local isolates. All patterns were reproducible and are now being used for typing M. bovis isolates. With either enzyme, the patterns produced for the M. tuberculosis, M. bovis, and M. africanum strains had many features in common, but all the M. bovis patterns were clearly more similar to each other than to the M. tuberculosis patterns. The patterns produced for the M. microti strain were markedly different from those produced for the other species. Restriction endonuclease analysis is clearly a useful method for inter- and intraspecific classifications of the tuberculosis complex.
Enzyme-linked immunosorbent assay of glycolipid antigens for identification of mycobacteria
Authors: Yanagihara D L , Barr V L , Knisley C V , Tsang A Y , McClatchy J K , Brennan P J
Published: January 2020

Enzyme-linked immunosorbent assays which are based on species- or type-specific glycolipids antigens and in which rabbit antisera are prepared with homologous strains are capable of distinguishing among serological variants of the Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum complex, Mycobacterium chelonei subspecies chelonei and abscessus, Mycobacterium simiae I and II, Mycobacterium kansasii, Mycobacterium szulgai, Mycobacterium xenopi, and Mycobacterium fortuitum biovariant peregrinum. The immunoreactive glycolipids can be divided into two classes. Those resistant to alkali, the C-mycoside glycopeptidolipids, are present in the M. avium-M. intracellulare-M. scrofulaceum, the M. chelonei subspecies chelonei and abscessus, and the M. simiae I and II complexes and in M. fortuitum biovariant peregrinum. The alkali-labile glycolipid antigens, the lipooligosaccharides, are present in M. kansasii, M. szulgai, and M. xenopi. In one study, the combination of enzyme-linked immunosorbent assay and alkaline susceptibility was compared with seroagglutination in the identification of 60 clinical isolates of nontuberculous mycobacteria: 45 showed perfect concordance, 9 could be narrowed to one, two, or three possibilities, and the rest did not correspond. In a second study involving 43 clinical isolates that were untypable by seroagglutination or were autoagglutinable, the results of enzyme-linked immunosorbent assay and thin-layer chromatography of glycolipid antigens were compared: 21 showed clear concordance. The results demonstrate that enzyme-linked immunosorbent assay is particularly useful in assessing the antigenicity of lipids, and sensitivity, ease, and rapidity recommend it as an adjunct to seroagglutination and thin-layer chromatography for the identification of nontuberculous mycobacteria.
Catalase and superoxide dismutase activities in virulent and nonvirulent Staphylococcus aureus isolates
Authors: Kanafani H , Martin S E
Published: January 2020

Catalase and superoxide dismutase (SOD) activities of virulent and nonvirulent isolates of Staphylococcus aureus were compared. The mean value of catalase activity for intact cell suspensions was 2,773 +/- 1,049 Kat f units (Kat f is defined as the ratio of the velocity constant of catalase at 0 min to the protein content in grams per milliliter); that of nonvirulent isolates was 154 +/- 92 Kat f units. The mean value of the catalase activities for lysates of virulent isolates was 260 +/- 120 Kat f units, and that of nonvirulent isolates was 31 +/- 19 Kat f units. Catalase levels in intact cells as well as in cell lysates were significantly different for virulent than for nonvirulent S. aureus isolates (P less than 0.001). The mean value of SOD activities was 20.85 +/- 11.48 U (1 U is defined as the amount of SOD required to inhibit the rate of reduction of cytochrome c by 50%) for virulent cell lysates, compared with a mean of 5.39 +/- 2.89 U for nonvirulent cell lysates. The SOD levels in virulent and nonvirulent isolates were significantly different (P less than 0.001). The virulence of the S. aureus isolates was determined by comparing weight gains of neonatal mice injected with virulent or nonvirulent strains. The percent weight gain of neonatal mice injected with virulent isolates was significantly lower than that of those injected with nonvirulent isolates.
Examination of uncommon clinical isolates of human adenoviruses by restriction endonuclease analysis
Authors: Hammond G W , Mauthe G , Joshua J , Hannan C K
Published: January 2020

Restriction endonuclease analysis was performed on reference strains of each unknown adenovirus subgenus, in comparison with 40 isolates not identified by our routine methods of neutralization with antisera of species 1 to 8. Several uncommon species would not have been identified initially without the assistance of reference laboratories (species 15, 35, and 37). Other species were identified by comparison with published adenovirus DNA restriction endonuclease patterns or from DNA analysis of reference strains (species 31, 40, and 41). Some isolates could not be matched beyond the level of presumptive adenovirus subgenus. Genomic DNA restriction endonuclease analysis of adenoviruses was useful for the identification of adenovirus isolates in a diagnostic virology laboratory. However, accurate interpretation of results will require more extensive DNA restriction endonuclease fragment analysis of a broader range of adenovirus species and genomic variant strains.
Survey in Vanuatu on enterotoxigenic Escherichia coli in children and infants with and without acute diarrhea
Authors: Germani Y , Montaville B , Fauran C , Brethes B
Published: January 2020

We have studied the incidence of enterotoxigenic Escherichia coli (ETEC) strains isolated from infants with and without diarrheal diseases in Vanuatu, South Pacific. Over a period of 5 months we have isolated enterotoxigenic E. coli strains from 29 (26.6%) of 109 children with acute diarrhea and from 13 (21.6%) of 60 children of the control group. In the group with diarrhea, 7 (6.4%) strains released heat-labile toxin, 7 (6.4%) released heat-stable toxin, and 15 (13.7%) produced both heat-labile and heat-stable toxin. In the control group, only one strain (1.6%) produced heat-stable toxin, 12 (20%) produced heat-labile toxin, and none produced both. Association of strains releasing heat-stable toxin or both heat-labile and heat-stable toxin with diarrhea was highly significant as shown by statistical analysis. The O serogroups and colonization factors CFA/I and CFA/II are presented.
Effect of adding sodium taurocholate to selective media on the recovery of Clostridium difficile from environmental surfaces
Authors: Buggy B P , Hawkins C C , Fekety R
Published: January 2020

The recovery of Clostridium difficile on a medium containing cefoxitin, cycloserine, fructose, and egg yolk was compared with that on media containing one of three preparations of sodium taurocholate. In aerobic environments contaminated with C. difficile, media containing either crude taurocholate from Mann Research Laboratories, New York, N.Y., or pure taurocholate from Sigma Chemical Co., St. Louis, Mo., recovered organisms significantly more often than did cefoxitin-cycloserine-fructose-egg yolk agar.
Survival of gram-negative and gram-positive bacteria artificially applied on the hands
Authors: Gontijo Filho P P , Stumpf M , Cardoso C L
Published: January 2020

We evaluated the survival of Pseudomonas aeruginosa, Klebsiella pneumoniae, Serratia marcescens, Escherichia coli, and Staphylococcus aureus, derived from either hospitalized patients or culture collections, on the fingertips of human volunteers. Over 99% of the bacteria died within 2 min of the application, and about 10(5) cells remained on the fingers for up to 90 min.
Enzyme immunoassay of serum beta-2-microglobulin levels in various histological forms of leprosy with special reference to its elevation in type I and type II lepra reactions
Authors: Saha K , Bhatnagar A , Sharma V K , Chakrabarty A K
Published: January 2020

The mean beta-2-microglobulin level in serum (3,362 +/- 2,494 micrograms/liter) for 76 leprosy patients, including 9 borderline-tuberculoid, 8 borderline-borderline, 9 borderline-lepromatous, and 16 lepromatous-lepromatous patients and 34 patients with type I or type II lepra reactions, was significantly higher (P less than 0.001) than that (2,122 +/- 1,844 micrograms/liter) for 35 normal subjects. It decreased significantly (P less than 0.001) as the disease glided down from borderline tuberculoid (3,173 +/- 899 micrograms/liter) to the lepromatous end (1,813 +/- 1,391 micrograms/liter). At the onset of type I or type II reaction, the mean beta-2-microglobulin level in serum increased (4,447 +/- 2,863 micrograms/liter), and it remained unchanged (4,433 +/- 2,623 micrograms/liter) after clinical remission. The beta-2-microglobulin level in serum decreased in 55.5% of the patients tested after subsidence of reaction. The level was significantly higher in patients with type II reactions (5,433 +/- 3,299 micrograms/liter) than in patients with type I reactions (3,558 +/- 2,171 micrograms/liter).
Plasmid DNA relatedness among different serogroups of Yersinia pseudotuberculosis
Authors: Ishiguro N , Nakaoka Y , Sato G , Tsubokura M
Published: January 2020

Thirteen different serogroup strains of Yersinia pseudotuberculosis and two strains of Yersinia enterocolitica O:3 were examined for the presence of plasmids and plasmid-mediated properties, calcium growth dependency, and autoagglutination. Two Y. enterocolitica strains and eight serogroup (IA, IIA, IIC, III, IVA, VB, VI, and VIII) strains, except for five serogroups (IB, IIB, IVB, VA, and VII), of Y. pseudotuberculosis harbored plasmids ranging in molecular size from 27 to 115 kilobases. Filter hybridization of restriction endonuclease-digested plasmid DNA from different serogroup strains indicated that all plasmid DNAs conferring calcium growth dependency and autoagglutination shared a high degree of DNA sequence homology, regardless of the different serogroups of Y. pseudotuberculosis and Y. enterocolitica.
Isolation of Acanthamoeba culbertsoni from a patient with meningitis
Authors: Lalitha M K , Anandi V , Srivastava A , Thomas K , Cherian A M , Chandi S M
Published: January 2020

A case of amoebic meningitis, presumably primary, was encountered in the Christian Medical College Hospital, Vellore, South India, in November 1983. The patient, a 40-year-old man, had cerebrospinal fluid rhinorrhea before the meningitis developed. Acanthamoeba culbertsoni was repeatedly demonstrated in and cultured from the cerebrospinal fluid. The patient responded dramatically to a combination therapy of penicillin and chloramphenicol.
Susceptibility of domestic cats to infections with Giardia lamblia cysts and trophozoites from human sources
Authors: Kirkpatrick C E , Green G A
Published: January 2020

The object of this study was to determine the importance of domestic cats in the epidemiology of human giardiasis. Of six laboratory-reared cats inoculated with cultured Giardia lamblia trophozoites from humans, only one showed the presence of cysts in the feces, and cysts were found on only 1 of the 80 days of observation. In a second experiment, eight cats were inoculated with G. lamblia cysts isolated from a human being. Over an 8-week period of observation, two of eight cats were found to have passed cysts in their feces, one on only one day and the other on 2 days. Postmortem examination of all of the cats found to be passing G. lamblia cysts at some time during the experiments did not reveal any small-intestinal trophozoites. These results suggest that domestic cats are relatively insusceptible to G. lamblia from humans and, consequently, that cats probably are not significant reservoir hosts of Giardia spp. infective for human beings. Moreover, it appears that the Giardia spp. which parasitize cats are distinct from those of human beings.
Rapid subtyping of equine herpesvirus 1 with monoclonal antibodies
Authors: Yeargan M R , Allen G P , Bryans J T
Published: January 2020

Two antigenically similar subtypes of equine herpesvirus 1 (EHV-1) cause disease in horses. A procedure for rapid differentiation of the two EHV-1 subtypes with monoclonal antibodies was developed. Subtype-specific pools of monoclonal antibodies were constructed, characterized, and used in enzyme immunofiltration and indirect immunofluorescence assays to subtype 50 epizootiologically unrelated field isolates of EHV-1. Both assays allowed accurate subtype identification of each EHV-1 isolate with the monoclonal antibody pools. The subtyping procedures were simple and amenable to typing many isolates at one time and permitted unambiguous EHV-1 subtype identification with 3 h after isolation of the virus in tissue culture.
Isolation of Inoue-Melnick virus and detection of antibodies in coded cerebrospinal fluid specimens from patients with disorders of the central nervous system
Authors: Melnick J L , Seidel E , Wang S S , Muchinik G
Published: January 2020

Inoue-Melnick virus was isolated from coded cerebrospinal fluid samples from 10 of 25 patients with chronic diseases of the central nervous system. Of 13 multiple sclerosis patients and 12 patients with a variety of other symptoms and signs 4 and 6, respectively, were positive. Replicate samples from the same patient yielded the same results, either positive or negative. Neutralizing antibody was present in cerebrospinal fluid at a dilution of 1:25 in the virus-positive patients. Only a single virus-negative patient was positive for antibody at this dilution.
Typing of heat-stable and heat-labile antigens of Campylobacter jejuni and Campylobacter coli by coagglutination
Authors: Wong K H , Skelton S K , Patton C M , Feeley J C , Morris G
Published: January 2020

A coagglutination system has been devised for typing heat-stable and heat-labile antigens of Campylobacter jejuni and C. coli. The use of protein A-positive Staphylococcus aureus cells carrying Campylobacter sp. serotype antibody and the treatment of Campylobacter sp. cells with DNase in the antigen suspension permitted rapid and specific coagglutination of rough (autoagglutinable) as well as smooth cultures. Cells of S. aureus were sensitized with Campylobacter sp. serotype antisera. Four to five types of sensitized S. aureus cells were pooled. A strain of Campylobacter sp. was first tested with the pools and then typed with the individual reagents of the reactive pool. After the described procedures, 68 serotype strains tested blindly as unknowns were correctly typed according to their heat-stable or heat-labile antigens. The two most commonly used typing schemes which are based separately on the heat-stable or the heat-labile antigens as assayed by passive hemagglutination and slide agglutination, respectively, can be utilized simultaneously in the coagglutination system for strain characterization. The coagglutination system is simple, yields results rapidly, conserves typing reagents, and offers the flexibility of formulating the pools of reagents according to the experimental design or the prevalence of serotypes in a geographic location. It should be a practical system for the typing of Campylobacter spp. in public health or clinical laboratories.
Cadmium chloride susceptibility, a characteristic of Campylobacter spp
Authors: Kazmi S U , Roberson B S , Stern N J
Published: January 2020

We report a simple diagnostic characteristic useful in the presumptive identification of Campylobacter jejuni and Campylobacter coli. Filter paper disks impregnated with cadmium chloride were placed on streaked agar medium. Zones of growth inhibition for Campylobacter spp. occurred at 1.25 micrograms per disk. Other enteropathogens (Salmonella spp., Shigella spp., Vibrio cholerae, Vibrio parahaemolyticus, Escherichia coli, and Yersinia enterocolitica) were resistant to at least 40 micrograms per disk, with the exception of a strain of Shigella flexneri, which showed first susceptibility at 10 micrograms per disk. Most of the 52 Campylobacter strains, which were isolated from human clinical and animal sources, showed zones of inhibition greater than 10 mm with 2.5 micrograms of cadmium chloride per disk. At 20 micrograms per disk, Campylobacter isolates from clinical sources were significantly (P less than 0.01) more susceptible to cadmium chloride inhibition than were those from meat samples.
"Campylobacter hyointestinalis" sp. nov.: a new species of Campylobacter found in the intestines of pigs and other animals
Authors: Gebhart C J , Edmonds P , Ward G E , Kurtz H J , Brenner D J
Published: January 2020

The name "Campylobacter hyointestinalis" sp. nov. is proposed for a Campylobacter species that was isolated from the intestines of pigs with proliferative enteritis. "C. hyointestinalis" is also found in the feces of cattle and has been isolated from the intestine of a hamster. "C. hyointestinalis" is distinguished from previously described catalase-positive Campylobacter species by colony morphology, ability to produce H2S in triple sugar iron agar, ability to grow anaerobically in 0.1% trimethylamine N-oxide hydrochloride, resistance to nalidixic acid, susceptibility to cephalothin and metronidazole, and hydrogenase activity. Sixteen "C. hyointestinalis" strains were highly related (greater than or equal to 76%) by DNA-DNA hybridization (hydroxyapatite method, 50 and 65 degrees C). Other Campylobacter species were less than or equal to 30% related to "C. hyointestinalis." The type strain of "C. hyointestinalis" is designated 80-4577-4 (= ATCC 35217), and its DNA has a guanine-plus-cytosine content of 36 mol%.
Evaluation of various rapid agglutination methods for the identification of Staphylococcus aureus
Authors: Baker J S , Bormann M A , Boudreau D H
Published: January 2020

A latex agglutination test (SeroSTAT Staph; Scott Laboratories, Fiskeville, R.I.) and two hemagglutination tests (Staphyloslide; BBL Microbiology Systems, Cockeysville, Md.; and Hemastaph; Remel, Lenexa, Kans.) were compared with the slide coagulase (SC) and tube coagulase (TC) tests at room temperature (22 to 25 degrees C) and at 37 degrees C for the rapid identification of Staphylococcus aureus. A total of 380 clinical strains of staphylococci were tested. The TC test performed at room temperature yielded the largest number of TC-positive results (n = 239), and based on this observation 239 organisms were classified as S. aureus and 141 were classified as non-S. aureus. The SC, TC (37 degrees C), SeroSTAT Staph, Staphyloslide, and Hemastaph tests correctly identified 210 (87.9%), 221 (92.5%), 238 (99.6%), 239 (100%), and 236 (98.7%) of the S. aureus isolates, respectively. Of the S. aureus isolates that were TC positive at room temperature 68% required 24 h of incubation before coagulase production was detected. There was one false-negative SeroSTAT Staph result and one false-negative Hemastaph result. The Staphyloslide test yielded two noninterpretable results (both organisms were later confirmed as non-S. aureus), whereas there were six noninterpretable results recorded with the Hemastaph test (four organisms were classified as non-S. aureus, and two were classified as S. aureus). The SeroSTAT Staph, Staphyloslide, and Hemastaph tests were all more sensitive than the conventional SC and TC (37 degrees C) tests and were considerably more rapid than the TC test at either temperature.
Evaluation of the RIM-N, Gonochek II, and Phadebact systems for the identification of pathogenic Neisseria spp. and Branhamella catarrhalis
Authors: Janda W M , Ulanday M G , Bohnhoff M , LeBeau L J
Published: January 2020

Methods for identifying Neisseria spp. include conventional and modified carbohydrate degradation procedures, chromogenic enzyme substrate tests, and immunologic coagglutination tests for Neisseria gonorrhoeae. In this study, we evaluated the abilities of the RIM-N carbohydrate degradation system (American MicroScan, Campbell, Calif.), the Gonochek II enzymatic identification system (Du Pont Co., Wilmington, Del.), and the Phadebact Gonococcus coagglutination test (Pharmacia Diagnostics, Piscataway, N.J.) to identify pathogenic Neisseria spp. and Branhamella catarrhalis. Both stock strains and clinical isolates, including 176 N. gonorrhoeae, 173 Neisseria meningitidis, 48 Neisseria lactamica, and 12 B. catarrhalis strains, were tested. The RIM-N identified 98% of the gonococci, 99% of the meningococci, 94% of the N. lactamica strains, and 100% of the B. catarrhalis strains within 1 h. The Gonochek II system identified 99% of the gonococci, 97% of the meningococci, 100% of the N. lactamica strains, and 100% of the B. catarrhalis strains within 30 min. Phadebact coagglutination provided clearly positive results for only 77% of the N. gonorrhoeae strains, producing negative or equivocal results with 23% of the strains. The RIM-N and Gonocheck II tests generally produced clear-cut reactions. An additional advantage of the Gonocheck II system was the small inoculum required for the performance of the test compared with the other systems, thus allowing the identification of N. gonorrhoeae directly from the primary isolation medium.
Chromogenic Limulus amoebocyte lysate assay for rapid detection of gram-negative bacteriuria
Authors: Nachum R , Berzofsky R N
Published: January 2020

A chromogenic Limulus amoebocyte lysate assay was evaluated as a rapid screening test for the detection of clinically significant gram-negative bacteriuria. The development of a distinctive yellow color after the addition of chromogenic substrate to the Limulus amoebocyte lysate-urine reaction mixture was used to measure greater than or equal to 10(5) gram-negative bacteria per ml. A total of 324 urine specimens were assayed, with 68 gram-negative urinary tract infections identified as defined by quantitative urine colony counts of greater than or equal to 10(5) bacteria per ml. Of these, 68 and 67 of 68 were detected by the chromogenic Limulus amoebocyte lysate assay at urine dilutions of 1:10 and 1:20, respectively. Nine false-positive chromogenic Limulus amoebocyte lysate assay results were observed at both urine dilutions and in the same specimens. At a urine dilution of 1:10, sensitivity and specificity were 100 and 96.6%, respectively, with predictive values of 100% for a negative test and 88.3% for a positive test. At a urine dilution of 1:20, sensitivity and specificity were 98.6 and 96.6%, respectively; predictive values were 99.6% for a negative test and 88.3% for a positive test. These data suggest that chromogenic Limulus amoebocyte lysate assay of urine has potential usefulness as a rapid, reliable, and easily performed and interpreted screening test for the diagnosis of clinically significant gram-negative bacteriuria.
Rapid herpes simplex virus detection in clinical samples submitted to a state virology laboratory
Authors: Mayo D R , Brennan T , Egbertson S H , Moore D F
Published: January 2020

Of 16,779 specimens received for herpes simplex virus (HSV) isolation since 1982, 4,465 (26.6%) were positive for HSV by either standard tissue culture or an antigen detection system (peroxidase-antiperoxidase; PAP). The overall isolation rate for genital vesicle specimens was lower (26.1%) than that for nongenital specimens (29.3%). Monthly isolation rates ranged from 19 to 32% for genital specimens and from 20 to 44% for nongenital specimens. Increasing demands for HSV isolation led to comparison of tissue culture with PAP. In the first comparison, HSV was isolated in single human fibroblast cell cultures from 1,019 of 4,261 specimens (23.9%), whereas single human fibroblast wells stained at 24 and 72 h postinoculation were PAP positive for 1,007 of 4,261 specimens (23.6%). In the second comparison, HSV was isolated from 225 of 1,026 (21.9%) specimens and duplicate human foreskin fibroblast cell wells stained at 24 and 72 h were PAP positive in 241 of 1,026 (23.5%). With the dual-well PAP system, all results were reported within 72 h, approximately 70% of positives were reported within 24 h, and considerable savings in time and materials resulted.