bluuebunny/crossref_metadata_2025 huggingface dataset:
Monoclonal antibodies to Bacteroides fragilis lipopolysaccharide
Authors: Linko-Kettunen L , Arstila P , Jalkanen M , Jousimies-Somer H , Lassila O , Lehtonen O P , Weintraub A , Viljanen M K
Published: January 2020

Monoclonal antibodies (MoAbs) to the lipopolysaccharide (LPS) of Bacteroides fragilis were produced by immunizing mice before hybridization with bacterial outer membranes solubilized with Triton X-100. Nineteen stabile clones were established. They all produced antibodies that reacted more strongly with purified B. fragilis LPS than with crude sonicated antigen in an enzyme immunoassay. Four MoAbs were studied by immunoblotting and enzyme immunoassay inhibition. Immunoblotting confirmed that the target of the MoAbs was LPS. Marked and homogeneous staining occurred in the immunoblotting with both purified LPS and outer membranes in the molecular weight range of 8,000 to 27,000. In enzyme immunoassay inhibition, MoAbs reacted positively with 93 to 96% of B. fragilis strains, including prototype strains ATCC 23745 and NCTC 9343. Within the B. fragilis group, the MoAbs reacted positively with two of five B. ovatus strains and two to six of nine B. thetaiotaomicron strains. No marked cross-reactivity with other bacteria was observed. These results confirm earlier findings that the B. fragilis LPS contains an immunodominant antigenic determinant common to almost all B. fragilis isolates.
Methicillin-resistant Staphylococcus aureus strains in New York City hospitals: inter-hospital spread of resistant strains of type 88
Authors: Schaefler S , Jones D , Perry W , Baradet T , Mayr E , Rampersad C
Published: January 2020

A survey of methicillin-resistant strains of Staphylococcus aureus received for phage typing indicated a marked increase of resistant strains received in 1982 and 1983. Of 62 hospitals in New York City which sent strains for phage typing, 35 had methicillin-resistant isolates. A significant development was the presence of strains of the same phage type at several hospitals, indicating a possible inter-hospital spread of these strains. Among strains present at several hospitals, the largest group was of experimental phage type 88. Strains of type 88 were received from 23 hospitals, representing 56% of all methicillin-resistant strains received from New York City hospitals. Strains of type 88 were resistant to all antistaphylococcal antibiotics, with the exception of vancomycin, and represented a major source of nosocomial infections at 13 hospitals. As experimental phage 88 is not routinely used for typing in U.S. laboratories, the nationwide distribution of strains of type 88 is difficult to assess.
Influence of growth medium on the in vitro activities of second- and third-generation cephalosporins against Streptococcus faecalis
Authors: Sahm D F , Baker C N , Jones R N , Thornsberry C
Published: January 2020

The influence of culture medium of the MICs of eight cephalosporins for 45 strains of Streptococcus faecalis was investigated. The MICs of cephalothin, cefamandole, and cefoperazone were not substantially influenced by the type of culture medium used. In contrast, MICs of cefuroxime, ceftizoxime, cefotaxime, cefmenoxime, and ceftriaxone varied markedly with both the commercial brand and the blood content of the broth used. The use of Mueller-Hinton broths (from Oxoid Ltd., GIBCO Diagnostics, and Difco Laboratories) supplemented with 5% lysed sheep blood frequently resulted in MICs that were greater than or equal to 16 times lower than the MICs obtained with these same broths without blood. Similar, but less marked, patterns were observed when supplemented and unsupplemented brain heart infusion and Sceptor broths were used. The influence of the broth on MICs suggests a complex interaction between some cephalosporins, medium components, and organisms. The cephalosporins that were affected by media share an identical moiety at the 7-acyl position (cefuroxime is slightly different), but this structure is not shared by those cephalosporins that were not affected. This commonality in structure at the 7-acyl position may be partially responsible for the observed results.
Recovery of Gardnerella vaginalis from blood by the quantitative direct plating method
Authors: La Scolea L J , Dryja D M , Dillon W P
Published: January 2020

This report describes a case of Gardnerella vaginalis sepsis in an obstetric patient whose blood cultures were negative by the conventional BACTEC system but positive by the quantitative direct plating method, which involves the direct inoculation of blood samples on chocolate agar and blood agar plates.
Endocarditis caused by Actinobacillus actinomycetemcomitans
Authors: Schack S H , Smith P W , Penn R G , Rapoport J M
Published: January 2020

Actinobacillus actinomycetemcomitans is a gram-negative coccobacillus which is a very rare cause of bacterial endocarditis. Preexisting cardiac lesions are a main contributing factor, and antibiotic prophylaxis has long been felt necessary before dental or other manipulation to prevent endocarditis. Penicillin in combination with an aminoglycoside has been the most often used treatment regimen. We present a case of endocarditis caused by this organism which developed after antibiotic prophylaxis for dental cleaning. Streptomycin and rifampin therapy resulted in the cure of the infection. The treatment and epidemiology of Actinobacillus endocarditis are reviewed.
Isolation of Brucella spp. from clinical specimens
Authors: Etemadi H , Raissadat A , Pickett M J , Zafari Y , Vahedifar P
Published: January 2020

For recovery of brucellae from clinical specimens, a lysis-concentration procedure was superior to the procedure of Castañeda (M. R. Castañeda, Proc. Soc. Exp. Biol. Med. 64:114-115, 1947).
Comparison of three tests for virulent Yersinia enterocolitica
Authors: Chang M T , Schink J , Shimaoka J , Doyle M P
Published: January 2020

Magnesium oxalate agar was found to select against virulent types of Yersinia enterocolitica. Small colonies isolated from magnesium oxalate agar which were presumably virulent generally contained no detectable virulence-specific plasmids, did not agglutinate with virulence-specific antiserum, and exhibited various degrees of virulence in mice. Results indicate that the virulence potential of a Y. enterocolitica isolate cannot always be identified by these in vitro methods and suggest that virulence also involves other factors yet to be identified.
Conversion of Blastomyces dermatitidis to the yeast form at 37 degrees C and 26 degrees C
Authors: Kane J
Published: January 2020

A partially defined agar medium, KT, has been developed and compared with brain heart infusion agar for the conversion of Blastomyces dermatitidis to the yeast form. On the KT medium, the mold form converted to a yeast form within 72 h of incubation at 37 degrees C or after 3 weeks at 26 degrees C. A nutritionally dependent dimorphism in B. dermatitidis was observed.
Laboratory identification of Haemophilus influenzae: effects of basal media on the results of the satellitism test and evaluation of the RapID NH system
Authors: Doern G V , Chapin K C
Published: January 2020

The effects of four different basal media, tryptic soy agar, brain heart infusion agar, nutrient agar, and Mueller-Hinton agar, were investigated with respect to the identification of Haemophilus influenzae with a satellitism test in which X and V growth factors were supplied by factor-impregnated filter paper strips. A total of 187 recent clinical isolates of H. influenzae were examined. Of these, 179 strains (95.7%) were correctly identified with tryptic soy agar, 173 (92.5%) with brain heart infusion agar, 105 (56.1%) with nutrient agar, and 133 (71.1%) with Mueller-Hinton agar. Failure to obtain a correct identification was usually the result of satelliting growth around V factor-containing strips, possibly due to the presence of trace amounts of hemin in the basal media, or was because of an absence of growth due to nutritional deficiencies in the basal media. All 187 H. influenzae strains were also examined with a new biochemical and chromogenic substrate micromethod, the RapID NH system (Innovative Diagnostics Systems, Inc., Decatur, Ga.). A total of 168 (89.8%) strains were correctly identified with this system.
Rapid procedure for isolation of plasmid DNA and application to epidemiological analysis
Authors: Takahashi S , Nagano Y
Published: January 2020

A rapid and simple plasmid isolation procedure was developed for the epidemiological analysis of plasmid-mediated antimicrobial resistance. By this method, plasmid DNAs ranging in molecular weight between 2.0 and 122 X 10(6) could be detected. Various bacteria, such as strains of the family Enterobacteriaceae, Pseudomonas aeruginosa, Haemophilus influenzae, and Staphylococcus aureus, could be analyzed. The plasmid DNA obtained could be directly used for restriction endonuclease analysis without further purification. In addition, this method made it possible to analyze several cultures at the same time.
Clinical laboratory comparison of the 10-ml isolator blood culture system with BACTEC radiometric blood culture media
Authors: Kellogg J A , Manzella J P , McConville J H
Published: January 2020

The efficiency of the 10-ml Isolator (E. I. du Pont de Nemours & Co., Inc.) for recovery of pathogens from blood was compared with that of BACTEC 6B and 7C media (Johnston Laboratories) by using 4,195 cultures from 1,662 patients. During the first phase of the study, BACTEC bottles were inoculated with 3 ml of blood; in the second phase, bottles were inoculated with 5 ml. The objectives were to compare results with similar blood volumes used for the detection of anaerobes as well as similar overall volumes and to determine the relative sensitivity of BACTEC media inoculated with the minimum and maximum volumes suggested by the manufacturer. From 180 patients, 391 significant isolates were recovered, 354 (91%) with the Isolator and 304 (78%) with the bottles. Isolators recovered 31 (15%) and 19 (18%) more pathogens overall than did the two-bottle system inoculated with 3 and 5 ml of blood, respectively, including 30 (36%) and 10 (34%) more Enterobacteriaceae. Recovery of anaerobes was greater in the BACTEC anaerobic medium, but only when its inoculum was increased to 5 ml. No significant differences existed between the two systems in pathogen detection times or detection of polymicrobic bacteremia. The Isolator contamination rate (8.3%) was approximately 4 times that of the bottles. The number of CFU of pathogen per milliliter of blood, blood volume sampled, and number of Isolators collected were more important than antimicrobial agent pretreatment in contributing to patient bacteremia of fungemia undetected by the Isolator. The Isolator appeared to be a practical alternative for recovery of aerobic and facultatively anaerobic pathogens from the blood.
Evaluation of the AutoMicrobic system Gram-Negative General Susceptibility-Plus Card
Authors: Van Horn K G , Vandernoot A M , Burke E W , McKitrick J C
Published: January 2020

The MICs of 10 antimicrobial agents were determined for 1,000 clinical aerobic and facultatively anaerobic gram-negative bacilli by the 6-h AutoMicrobic system Gram-Negative General Susceptibility-Plus Card (GSC+) (Vitek Systems, Inc., Hazelwood, Mo.) and by an 18-h reference agar dilution method. Results obtained by both systems were evaluated for twofold dilution discrepancies and for interpretive category discrepancies. MICs that differed by more than one twofold dilution between the two test systems were considered to be discrepant. The overall twofold dilution agreement of the GSC+ MICs to agar dilution MICs was 94.0%. The agreement for each drug tested ranged from 91.7% (ampicillin and chloramphenicol) to 96.5% (carbenicillin). Interpretive categories were considered in agreement when the susceptibility interpretations obtained by the two systems were both very susceptible, moderately susceptible, or resistant. The overall interpretive category agreement was 80.5%. This agreement increased to 95.8% when all minor errors (moderately susceptible, one system; very susceptible, the other system) that would not affect therapy were omitted. The AutoMicrobic system GSC+ provides rapid and accurate susceptibility test results for clinical aerobic and facultatively anaerobic gram-negative bacilli.
Modified enzyme immunoassay for detecting Neisseria gonorrhoeae antigens
Authors: Papasian C J , Bartholomew W R , Amsterdam D
Published: January 2020

A commercial modification of an enzyme immunoassay (EIA) (Gonozyme; Abbott Laboratories, North Chicago, Ill.) for detection of Neisseria gonorrhoeae antigens was compared with conventional culturing. Specimens from males and females were collected at a sexually transmitted disease clinic; additional female specimens were collected at an obstetrics and gynecology clinic. EIA sensitivity and specificity for males were 100 and 98.6%, respectively (68 negative, 34 positive, 1 false-positive, and 0 false-negative). EIA sensitivity and specificity for female sexually transmitted disease clinic patients were 74.4 and 95.7%, respectively (66 negative, 29 positive, 3 false-positive, and 10 false-negative) EIA sensitivity and specificity for obstetrics and gynecology clinic patients were 100 and 99.2%, respectively (6 positive, 119 negative, 1 false-positive, and 0 false-negative). In female patients from whom multiple swab specimens were collected, the sequence of specimen collection and subsequent EIA analysis affected sensitivity.
Bacteriuria screening by direct bioluminescence assay of ATP
Authors: Schifman R B , Wieden M , Brooker J , Chery M , Delduca M , Norgard K , Palen C , Reis N , Swanson J , White J
Published: January 2020

A direct bioluminescence assay for bacteriuria screening is described and compared with the MS-2 system (Abbott Laboratories, Irvine, Tex.) and the chemical strip, Gram smear, and calibrated-loop methods. A total of 973 specimens were tested. Unlike previously described bioluminescence methods, this test measures total ATP in urine without pretreatment of samples to remove somatic ATP. The result was compared with an ATP standard (20 ng/ml). A low result (less than 3% of standard) was interpreted as negative and a high result (greater than 10% of standard) as positive. Samples with intermediate results (38% of total) were incubated at 35 degrees C in thioglycolate broth (1:10). A 2% increase in ATP concentration was interpreted as positive. The sensitivity of this method for detecting greater than 10(5) pathogens per ml was 92.3% and was comparable to those of the MS-2 system (92.7%) and the Gram smear method (90.5%). The chemical strip method was less sensitive (84.0%). The direct bioluminescence method was more sensitive than were the MS-2 system and the Gram smear method for detecting low-level bacteriuria (less than 10(3) to 10(5) organisms per ml), primarily because of associated pyuria. Thioglycolate broth provided a suitable medium for ATP production, and 5% CO2 decreased bacterial ATP synthesis during log-phase growth. The direct bioluminescence assay is rapid, simple, cost-effective, and reliable for bacteriuria screening.
Comparison of indirect hemagglutination and rapid plate agglutination tests with counterimmunoelectrophoresis for typing Pasteurella haemolytica
Authors: Chengappa M M , Carter G R , Bailie W E
Published: January 2020

A rapid, simple, and accurate counterimmunoelectrophoresis (CIE) technique was developed and compared with the indirect hemagglutination and rapid plate agglutination techniques for serotyping cultures of Pasteurella haemolytica. The CIE test had 100% correlation with the conventional indirect hemagglutination test and, after serum absorption, correctly identified cultures representing the 12 established serotypes and 49 field isolates of P. haemolytica with reasonable rapidity. Cross-reactions were observed in the CIE and rapid plate agglutination tests but not in the indirect hemagglutination test with antisera prepared from the 12 established serotypes. These cross-reactions were eliminated from the CIE test but not from the rapid plate agglutination test by absorption of antisera with cells which possessed the cross-reacting antigens. Avian isolates of P. haemolytica did not type with antisera to the 12 established serotypes by any of the methods. Both homologous and heterologous reactions were observed with these strains in the rapid plate agglutination and CIE tests with antisera prepared from six selected cultures. These results support the previous finding that the taxonomic relationship of these avian strains to P. haemolytica is questionable.
Laboratory experience with radiometric detection of bacteremia with three culture media
Authors: Wicher K , Koscinski D
Published: January 2020

In two long-term studies, the BACTEC radiometric system for detection of bacteremia was evaluated with three culture media each: (i) BACTEC media 6A (for aerobes) and 7B (for anaerobes) plus a thioglycolate medium and (ii) BACTEC media 6A, 7B, and 8A (hypertonic). In study 1, clinically significant isolates were identified in 1,873 (13.9%) of 13,432 blood cultures with all three media. The thioglycolate medium revealed 143 (1.1%) organisms not recovered from the 6A and 7B media. In study 2, isolates were identified in 1,135 (12.9%) of 8,759 cultures with all three media; 104 (1.2%) organisms were isolated only from the hypertonic medium. The increased yield of positive cultures in the three-medium system is likely due to the larger volume of blood cultured.
Biotypes of Gardnerella vaginalis
Authors: Piot P , Van Dyck E , Peeters M , Hale J , Totten P A , Holmes K K
Published: January 2020

A simple and reproducible scheme for identifying biotypes of Gardnerella vaginalis has been developed, based on reactions for lipase, hippurate hydrolysis, and beta-galactosidase. Among a total of 359 strains tested, eight biotypes were observed, the most common ones being types 1 (beta-galactosidase positive, lipase positive, hippurate positive), 2 (beta-galactosidase negative, lipase positive, hippurate positive), and 5 (beta-galactosidase negative, lipase negative, hippurate positive). The distribution in biotypes was similar among isolates from Antwerp, Seattle, and Nairobi. There were no differences in biotypes between strains isolated from patients with and without bacterial vaginosis (nonspecific vaginitis). Up to 14% of women with bacterial vaginosis harbored at least two different biotypes of G. vaginalis in the vagina. G. vaginalis strains isolated before and after treatment for bacterial vaginosis belonged to identical biotypes when the time interval between two specimens was less than 1 week. Similarly, G. vaginalis isolates from the vaginas of women with bacterial vaginosis and from the urethras of their male sex partners belonged to identical biotypes when strains were isolated within the same 24-h period from both partners (P less than 0.005).
Evaluation of Gonochek-II as a rapid identification system for pathogenic Neisseria species
Authors: Welborn P P , Uyeda C T , Ellison-Birang N
Published: January 2020

The Gonochek-II test kit (E-Y Laboratories, San Mateo, Calif.) may be used to rapidly identify (within 30 min) Neisseria gonorrhoeae, Neisseria meningitidis, Neisseria lactamica, and Branhamella catarrhalis. The kit consists of oxidase swabs, chromogenic substrates, and EY-20 reagent (diazonium salt derivative). A beta-lactamase tube is also included. Fifty-two clinical isolates were tested from cervical (22 isolates), urethral (13), throat (11), rectal (5), and urine (1) sources. All strains were oxidase-positive, gram-negative diplococci isolated on Thayer-Mayer plates incubated at 35 degrees C in 5% CO2 for 18 to 24 h. Each strain was tested by Gonochek-II, RapID NH (Innovative Diagnostics, Decatur, Ga.), and conventional rapid carbohydrate utilization. Forty-four isolates of N. gonorrhoeae and six N. meningitidis were identified. Only two isolates tested (4%) were identified as species other than these two. Gonochek-II outperformed all other methods for identification of N. gonorrhoeae from cervical and urethral areas. N. meningitidis strains were correctly identified in all cases. Ease and rapidity of the procedure, coupled with a small inoculum requirement and reliable results, led us to favor Gonochek-II for routine identification of pathogenic Neisseria species.
Use of bone marrow fibroblasts to prepare targets for an HLA restricted-cytotoxicity assay system
Authors: Bowden R A , McGavren L , Hayward A R , Levin M J
Published: January 2020

Methods are described for obtaining and growing fibroblasts from bone marrow for use as virus-infected targets. Fibroblasts obtained at the time of routine marrow examination were maintained through 12 to 18 passages as confluent monolayers. Bone marrow fibroblasts could be infected with varicella-zoster virus, and these infected cells were suitable targets for a 51Cr-release cytotoxicity assay. Since these virus-infected cells retain their HLA-A and -B antigens, they are readily available to study the immune cells which mediate virus-specific cytotoxicity.
Evaluation of Gonozyme, an enzyme immunoassay for the rapid diagnosis of gonorrhea
Authors: Manis R D , Harris B , Geiseler P J
Published: January 2020

A new indirect enzyme immunoassay (EIA), Gonozyme (Abbott Laboratories), was assessed for rapid detection of gonococcal antigens. A correlation of optic density (OD) readings by EIA with colony counts of serial dilutions of Neisseria gonorrhoeae ATCC 19424 disclosed that EIA detected 10(3) CFU/ml at OD readings of 0.1 to 0.3, that EIA consistently detected greater than or equal to 10(4) CFU/ml at OD readings of 0.6 to 1.3, and that concentrations of greater than or equal to 10(5) CFU/ml were associated with OD readings of greater than or equal to 2.0. The clinical usefulness of Gonozyme was evaluated by comparing results of EIA with those of Gram stain (GS) and culture for N. gonorrhoeae from urethral and endocervical swabs obtained prospectively in 886 randomly selected patients attending a clinic for sexually transmitted diseases. The patients evaluated included 83 female contacts of men with gonorrhea and 56 patients seen at the clinic for test of cure. In tests with 295 males, the sensitivities of GS and EIA were 91.3 and 97.1%, respectively, and both tests had specificities of greater than 96%. In tests with 591 females, the sensitivities of GS and EIA were 51.4 and 96.4%, respectively (P less than 0.0001, Z proportionality test), and the specificities were 98.7 and 86.5%, respectively (P less than 0.0001). In tests with 61 females and 3 males, EIA was positive, whereas GS and cultures were negative for N. gonorrhoeae. Gonozyme is a highly sensitive method for rapid detection of gonococcal antigens. EIA is comparable to GS for males and more sensitive though less specific than GS for females. Possible reasons for the lower specificity of EIA for females are discussed. Due to its high negative predictive value for female contacts, EIA offers an alternative to epidemiological treatment of contacts before culture results.
Evaluation of passive bacterial agglutination for the diagnosis of typhoid fever
Authors: John T J , Sivadasan K , Kurien B
Published: January 2020

We evaluated the reliability of a passive bacterial agglutination test to detect Salmonella typhi somatic antigen(s) in the sera of patients with typhoid fever. It was positive in 32 of 33 bacteriologically proven typhoid fever cases. Among 13 patients with a presumptive diagnosis of typhoid fever, 11 were positive by passive bacterial agglutination. The serum of one patient with paratyphoid A was also positive. Among 50 febrile patients without typhoid fever, one was persistently positive during the course of illness; 49 were negative. The sensitivity, specificity, and accuracy indices of the passive bacterial agglutination test were over 95%. The positive and negative predictive values were 94 and 98%, respectively.
Comparative evaluation of the Eiken and API 20E systems and conventional methods for identification of members of the family Enterobacteriaceae
Authors: Castillo C B , Bruckner D A
Published: January 2020

To evaluate the accuracy and utility of the Eiken Systek No. 1 (Eiken system; Eiken Chemical Co., Ltd., Tokyo, Japan), we conducted a clinical comparison, with 345 Enterobacteriaceae isolates, of the Eiken System with API 20E (Analytab Products, Inc., Plainview, N.Y.) and conventional methods. The Eiken system is a 21-biochemical-test battery tray stored at 25 degrees C and inoculated in one step. It is similar to the API 20E except that the Eiken system contains malonate, adonitol, and maltose; lacks gelatin, sucrose, melibiose, amygdalin, and arabinose; and uses reagent strips instead of liquid reagents. The API 20E and Eiken systems correctly identified 339 (97.7%) and 276 (79.5%), respectively, and misidentified 3 (0.9%) and 13 (3.7%), respectively, of the isolates. There were no identification codes for 5 (1.4%) organisms with the API 20E and 58 (16.7%) organisms with the Eiken system; of these latter unidentified organisms, 42 were identified as Proteus spp., Morganella sp., and Providencia rettgeri by conventional methods. There was no significant difference between the two rapid systems in total time required for inoculation and reading. Modifications for interpretation of decarboxylase and oxidase tests were needed for the Eiken system, and manipulation of reagent strips required considerable dexterity. However, the Eiken system was easier to inoculate than the API 20E, and, with minor increases in the data base to include more of the Proteus and Morganella spp. and P. rettgeri, the system should be reliable for identification of members of the family Enterobacteriaceae.
Detection of immunoglobulin M and G antibodies against cytomegalovirus early and late antigens by enzyme-linked immunosorbent assay
Authors: Middeldorp J M , Jongsma J , ter Haar A , Schirm J , The T H
Published: January 2020

A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) is described for the detection of immunoglobulin M and antibodies with specifity for human cytomegalovirus (CMV) early (CMV-EA) and late (CMV-LA) antigens. The emphasis is on the production of high-quality CMV antigens, CMV-EA and CMV-LA separately, and conditions for their application in the ELISA. The induction of CMV-EA and -LA in infected cell extracts was studied in detail by using human sera with defined antibody specificity for CMV-EA and CMV-LA. This resulted in the development of a simple whole cell extraction procedure that provided a high yield of CMV antigens with reproducible antigen quality. The antigens were specific for the detection of anti-CMV antibodies. The influence of autoantibodies on the determination of CMV-specific antibodies was investigated. Parallel analysis of 322 human sera by indirect immunofluorescence and ELISA showed a high correlation between both assays (r = 0.9674 for CMV-EA and 0.9362 for CMV-LA). Antibody titers determined by ELISA were equal to (for CMV-EA) or slightly higher (for CMV-LA) that those determined by immunofluorescence but significantly higher (20- to 5,120-fold) than those determined by complement fixation. From 191 sera positive by ELISA (titer greater than or equal to 40) 4 (2.1%) were negative by immunofluorescence (titer less than 40), and from 61 ELISA-positive sera 12 (19.6%) were negative (titer less than 8) when tested by complement fixation. Consequently, ELISA for CMV may prove to be more reliable for the selection of CMV-seronegative blood donors than these other methods. The use of high-quality antigens allows more economic handling of large-scale serum determinations. Possibilities for further automation are discussed.
Immunoglobulin M antibody capture enzyme-linked immunosorbent assay for diagnosis of St. Louis encephalitis
Authors: Monath T P , Nystrom R R , Bailey R E , Calisher C H , Muth D J
Published: January 2020

Sera from patients with St. Louis encephalitis were tested with an immunoglobulin M (IgM) antibody capture enzyme immunoassay (MAC ELISA). The assay used five reagents: antihuman IgM, test serum, sucrose-acetone-extracted mouse brain antigen, broadly cross-reactive flavivirus monoclonal antibody conjugated to alkaline phosphatase, and substrate (p-nitrophenyl phosphate). MAC ELISA endpoint titers correlated (r = 0.893) with the absorbance value of a 1:100 dilution of patient serum. Significant (fourfold or greater) changes in the endpoint titers between paired sera corresponded to a critical ratio (ratio of absorbance values at the 1:100 dilution) of greater than or equal to 1.3. IgM antibodies were detected in 71% of patients bled at 0 to 3 days after the onset of illness, in 99% bled at 4 to 21 days, and in 91% bled at 22 to 67 days. Thereafter, the IgM seropositivity rate declined; however, 29% of sera were still positive at 115 to 251 days after the onset of illness. MAC ELISA titers were significantly correlated with hemagglutination inhibition (r = 0.258) and neutralization (r = 0.711) titers. Because IgM antibodies appeared early and waned rapidly, a diagnosis was made on the basis of a decrease in titer more often by MAC ELISA than by hemagglutination inhibition, complement fixation, or neutralization tests. IgM antibodies generally showed a high degree of specificity; heterologous cross-reactions were, however, present in 4 of 14 sera examined. The MAC ELISA is useful for the rapid, early diagnosis of St. Louis encephalitis.
Antimicrobial agent susceptibility patterns of bacteria in hospitals from 1971 to 1982
Authors: Atkinson B A , Lorian V
Published: January 2020

Bacterial susceptibility to 16 commonly used antibiotics was analyzed for a 12-year period (from 1971 to 1982, inclusive). Susceptibilities of 5,828,243 strains isolated from a mean of 242 hospitals nationwide and of 194,575 strains isolated at the Massachusetts General Hospital, Boston, Mass., and the Bronx Lebanon Hospital Center, New York, N.Y., were compared. Strains of Escherichia coli, Staphylococcus aureus, Haemophilus influenzae, and Pseudomonas aeruginosa showed virtually the same susceptibilities to antibiotics throughout the 12-year period, whereas Streptococcus faecalis and Staphylococcus epidermidis showed significant increases in resistance to most antibiotics. The close similarity between antibiotic susceptibilities shown at both the 242 hospitals and the 2 individual hospitals suggests that this analysis accurately reflects trends of bacterial resistance to antibiotics in U.S. hospitals. Since most of the species analyzed produce serious disease and high mortality, their susceptibility to antibiotics is relevant both to physicians treating infectious diseases and to epidemiologists.
Quality control of susceptibility tests with 5-micrograms trimethoprim disks
Authors: Barry A L , Jones R N , Gavan T L
Published: January 2020

In 1981 and again in 1984, we performed multi-laboratory studies to develop quality control limits for susceptibility tests with 5-micrograms trimethoprim disks. Zone size limits of 21 to 28 mm are recommended for tests with Escherichia coli ATCC 25922, and limits of 19 to 26 mm are recommended for tests with Staphylococcus aureus ATCC 25923. For screening Mueller-Hinton agar, tests with Streptococcus faecalis ATCC 33186 or ATCC 29212 are recommended: zones should be fairly clear and greater than or equal to 22 mm (trimethoprim disks) or greater than or equal to 24 mm (trimethoprim-sulfamethoxazole disks).
Nitrocellulose filter blots for species identification of Mobiluncus curtisii and Mobiluncus mulieris
Authors: Roberts M C , Hillier S L , Schoenknecht F D , Holmes K K
Published: January 2020

Seventy strains of Mobiluncus, motile curved anaerobic bacteria associated with bacterial vaginosis, were correctly identified to species level by using bacteria fixed to nitrocellulose and hybridized with 32P-labeled DNA.
Prevalence and laboratory identification of methicillin-resistant Staphylococcus aureus in community hospitals
Authors: Mahoney M C , Eckman M R , Stolee T A , Cossalter D J
Published: January 2020

The prevalence of methicillin-resistant Staphylococcus aureus infections in community hospitals in northern Minnesota, Wisconsin, and Michigan was found to be one case in 82,565 patients. The percentage of S. aureus isolates resistant to methicillin was less than 0.2% (5 of 2,835). In this study, conducted from 1 June 1982 to 31 May 1983, a laboratory-controlled methodology was used.
Application of "Hirt supernatant" DNA to the molecular epidemiology of cytomegalovirus infections
Authors: Eizuru Y , Inagawa S , Minamishima Y
Published: January 2020

Human cytomegalovirus DNA was isolated from infected cells by the Hirt method (B. Hirt, J. Mol. Biol. 26:365-369, 1967). The restriction endonuclease cleavage patterns of DNA obtained in this manner were comparable with those of DNA extracted from purified virions. The "Hirt supernatants" were satisfactory for identifying individual cytomegalovirus strains by their DNA fingerprinting.
Detection of group B streptococcal antigen in body fluids by a latex-coupled monoclonal antibody assay
Authors: Rench M A , Metzger T G , Baker C J
Published: January 2020

A commercially available latex agglutination reagent, Directigen (Hynson, Westcott & Dunning, Div. Becton Dickinson & Co., Baltimore, Md.), in which a murine monoclonal antibody to group B streptococcal (GBS) antigen is the active component, was evaluated by using body fluid specimens from 94 sick infants. Antigen was detected in one or more admission specimens from 18 of 19 (94.7%) infants with symptomatic GBS infection. In 15 patients with GBS meningitis, cerebrospinal fluid, serum, and 10-fold-concentrated urine specimens were positive in 87, 50, and 100%, respectively. GBS antigen was detected in 50% of unconcentrated urines, but in no sera from infants with nonmeningitic GBS sepsis. Among specimens from 27 infants with invasive infection due to organisms other than GBS and from 48 culture-negative sick infants, false-positive latex agglutination reactions occurred in only 4 (9.5%) urine specimens and in no cerebrospinal fluid or serum specimens. The 94.7% sensitivity and specificity of the Directigen GBS test indicate that it is a useful reagent for the rapid diagnosis of invasive GBS infection in young infants.
Cryptosporidium spp., a frequent cause of diarrhea in Central Africa
Authors: Bogaerts J , Lepage P , Rouvroy D , Vandepitte J
Published: January 2020

Cryptosporidium oocysts were present in 20 (10.4%) of 193 Rwandese children and in 3 (3.0%) of 100 adults with diarrhea. In four of the children and in one adult, Cryptosporidium was associated with other enteric pathogens. The higher incidence of Cryptosporidium in diarrheic children was statistically significant. The parasite was not found in 94 formed stools submitted for parasitological examination. The mean age of the Cryptosporidium-positive children was 13.3 months. In four children, Cryptosporidium was associated with severe malnutrition. All of those required rehydration, and one child died as a direct consequence of severe diarrhea. The three adult patients showed no recognizable immunodeficiency, and their diarrhea resolved spontaneously. Staining with 1% safranin was not only more simple and rapid but also more sensitive than the modified Ziehl-Neelsen technique.
Use of the directigen latex agglutination test for detection of Haemophilus influenzae, Streptococcus pneumoniae, and Neisseria meningitidis antigens in cerebrospinal fluid from meningitis patients
Authors: Sippel J E , Hider P A , Controni G , Eisenach K D , Hill H R , Rytel M W , Wasilauskas B L
Published: January 2020

Cerebrospinal fluid specimens from 257 persons were tested for the presence of bacterial antigens by counterimmunoelectrophoresis and the Directigen meningitis test (Hynson, Westcott & Dunning, Div. Becton Dickinson & Co., Baltimore, Md.). The specimens were obtained from 162 patients with meningitis caused by Haemophilus influenzae type b, Streptococcus pneumoniae, or Neisseria meningitidis serogroups A and C and from 95 patients without bacterial meningitis or meningitis caused by other bacterial agents. Directigen detected H. influenzae type b antigen in 83% (69 of 83) of the specimens obtained from patients with H. influenzae disease, pneumococcal antigen in 77% (30 of 39) of the specimens from patients with pneumococcal disease, and N. meningitidis antigen in 93% (37 of 40) of the specimens from patients with disease caused by N. meningitidis serogroups A and C. The comparable figures for counterimmunoelectrophoresis were 66% (55 of 83), 79% (31 of 39), and 78% (31 of 40), respectively. No false-positive reactions were reported with the Directigen reagents. Nonspecific reactions (agglutination with more than one of the four Directigen latex reagents) were noted with five specimens. The nonspecific reactions were resolved in four of the five specimens by heating (100 degrees C for 3 min). The accumulated data demonstrate that the sensitivity of the Directigen meningitis test is better than or at least equivalent to the sensitivity of counterimmunoelectrophoresis for the detection of antigens in cerebrospinal fluid.
Importance of culture in laboratory diagnosis of Bordetella pertussis infections
Authors: Gilligan P H , Fisher M C
Published: January 2020

Nasopharyngeal secretions from 223 patients were examined for the presence of Bordetella pertussis and B. parapertussis by culturing on Regan-Lowe agar (J. Regan and F. Lowe, J. Clin. Microbiol. 6:303-309, 1977) and by direct fluorescent-antibody testing. B. pertussis was found in 38 patients; B. parapertussis was recovered from 2. Culturing was more sensitive (38 of 38 patients) than direct fluorescent-antibody testing (26 of 38 patients) for the detection of B. pertussis. Overgrowth by other organisms (7 of 223 patients) was uncommon. The patients with B. pertussis infections were generally less than 1 year old, had received no or one immunization, and had coughing spells but infrequently had whooping cough. Accurate diagnosis of B. pertussis infections should include culturing.
Collaborative evaluation of the Abbott Avantage system for identification of frequently isolated nonfermentative or oxidase-positive gram-negative bacilli
Authors: Jorgensen J H , Dyke J W , Helgeson N G , Cooper B H , Redding J S , Crawford S A , Andruszewski M T , Prowant S A
Published: January 2020

The capability of the Abbott Avantage system to identify 10 species of commonly isolated glucose nonfermentative or oxidase-positive gram-negative bacilli in a 5-h test period was evaluated in a collaborative study. The Avantage nonenteric data base uses 20 biochemical test reactions performed in an expanded Abbott bacterial identification cartridge plus the results of a manual oxidase test. The species included in the Avantage data base are Acinetobacter anitratus, Acinetobacter Iwoffi, Aeromonas hydrophila, Flavobacterium meningosepticum-IIb group, Pseudomonas aeruginosa, Pseudomonas cepacia, Pseudomonas fluorescens-putida group, Pseudomonas maltophilia, Pasteurella multocida, and Plesiomonas shigelloides. The collaborative study included the testing of 200 coded challenge strains in all three laboratories and the subsequent testing of an additional group of 100 to 200 clinical isolates recovered independently by each laboratory. Reference identifications for all isolates were determined by conventional biochemical test reactions. The overall accuracy of identification of the coded challenge strains for the three laboratories was 97%, whereas 95% of 437 clinical isolates and selected stock cultures of clinical derivation were identified correctly.
Laboratory and clinical evaluation of modified New York City medium (Henderson formulation) for the isolation of Neisseria gonorrhoeae
Authors: Anstey R J , Gun-Munro J , Rennie R P , Thornley J H , Schaus D G , Lannigan R , Hussain Z , Maharajah R S
Published: January 2020

The growth-promoting properties of several ingredients of New York City (NYC) medium were investigated by using 100 recently isolated clinical strains of Neisseria gonorrhoeae. Difco GC medium base promoted the growth of large colonies of gonococci significantly better than other commercial GC medium bases. A 1% agar concentration resulted in the growth of larger colonies than 2% agar, without affecting the stability of the gel. Neither replacement of horse plasma with horse serum nor reducing the concentration of added serum from 12 to 3% affected the growth of even the most fastidious strains tested. From these observations, a modified NYC medium (Henderson) has been formulated which is easy to prepare and less expensive than NYC or Thayer-Martin medium. In a direct clinical comparison with 1,275 specimens, an isolation rate of 6.5% was achieved with both NYC medium and the Henderson formulation.
Clinical evaluation of precipitin tests for genital actinomycosis
Authors: Persson E , Holmberg K
Published: January 2020

A precipitin test system for antibodies against Actinomyces israelii, comprising a combination of counterimmunoelectrophoretic and crossed immunoelectrophoretic gel techniques, was evaluated for its clinical usefulness in diagnosing genital actinomycosis. A total of 263 serum samples from healthy women and women with proven actinomycosis, A. israelii-associated salpingitis, other gynecological infections, or miscellaneous gynecological diseases were analyzed. Six different precipitins could be detected. Five precipitins were defined as specific for actinomycosis, whereas one was found to occur nonspecifically in women with gynecological infections. The specificity of the test system for the detection of cases of genital actinomycosis was 98%, and the sensitivity was 83%. The accuracy was 100% for negative prediction and 45% for positive prediction. Thus, the test was shown to be valuable for a noninvasive diagnosis of genital actinomycosis.
Prevalence of gram-negative bacilli in nares and on hands of pharmacy personnel: lack of effect of occupational exposure to antibiotics
Authors: Weil D C , Chou T , Arnow P M
Published: January 2020

Exposure to antibiotics alters host flora and facilitates colonization by gram-negative bacilli (GNB). This may be important among pharmacy personnel, who have frequent contact with antibiotics and who have sometimes been suspected of inadvertently introducing GNB into parenteral solutions during admixture. We evaluated the risk of colonization by GNB, especially by tribe Klebsielleae (TK) which can proliferate in intravenous fluids, by culturing the hands and nares of 98 pharmacy personnel and 56 control subjects. Four culture surveys of pharmacy personnel yielded mean isolation rates of 79 and 52% for GNB and TK, respectively, from hands and 12 and 6.7% for GNB and TK, respectively, from nares; these rates did not differ significantly from those for control subjects (P greater than 0.1). The frequency with which pharmacy personnel performed antibiotic admixture did not significantly affect the rate of isolation of GNB or TK (P greater than 0.2). No multiresistant strains were isolated, and susceptibility patterns were similar for GNB species from pharmacy personnel and controls. These data indicate that occupational exposure of pharmacy personnel to antibiotics is not of sufficient magnitude to increase rates of nasal colonization or hand contamination with GNB.
Carrier rate of Streptococcus suis capsular type 2 in palatine tonsils of slaughtered pigs
Authors: Arends J P , Hartwig N , Rudolphy M , Zanen H C
Published: January 2020

Palatine tonsils of 143 slaughtered pigs aged 4 to 6 months were investigated for the presence of Streptococcus suis type 2. Slices (50 micron) of frozen tonsils were cultured on a selective agar medium containing antibodies against S. suis type 2 in which colonies of this bacterium showed a halo of immunoprecipitation. When tonsils were sectioned in one plane S. suis type 2 was found in 45 of 143 pigs (32%). This percentage increased to 50% when tonsils were sectioned in more then one plane, which was done on 55 tonsils. The first 45 strains showing a ring of immunoprecipitation were studied and found to be biochemically identical to our reference strain 735 (de Moor) and to 23 isolates from human patients with meningitis. In slices incubated for 24 h at 37 degrees C on selective agar plates and stained with hematoxylin and eosin after fixation, it could be demonstrated that S. suis type 2 was confined to the crypt lumen. The same was true in sections fixed directly (without incubation) that were stained by an indirect immunoperoxidase method with a rabbit anti-S. suis type 2 serum.
Comparison of nonspecific reactivity in indirect and reverse immunoassays for measles and mumps immunoglobulin M antibodies
Authors: Tuokko H
Published: January 2020

Serum specimens collected from patients convalescing from acute measles or mumps infections, other viral infections, or rheumatoid arthritis and from blood donors were tested in indirect and reverse assays for measles and mumps immunoglobulin M (IgM) antibodies. All the samples from patients convalescing from acute mumps and measles infections gave positive IgM results in both tests. However, 6% of sera from patients recovering from other viral infections, 68.4% of sera from patients with rheumatoid arthritis, and 5.6% of sera from normal blood donors gave false-positive results by the indirect measles IgM enzyme immunoassay (EIA). By the indirect mumps IgM EIA, 9% of sera from other viral infections, 70.1% of sera from patients with rheumatoid arthritis, and 5.6% of sera from normal blood donors gave false-positive reactions. The reverse test system for measles IgM gave false-positive results in 1.5% of sera from the group with other viral infections, and the reverse mumps EIA gave false-positive results in 0.9% of the patients. Other sera groups did not react in either measles or mumps reverse IgM assays. The results indicated that although nonspecific reactions are frequent in indirect IgM tests for viral antibodies, such reactions are rarely encountered when reverse IgM EIA tests are employed.
Improved biotyping schemes for Campylobacter jejuni and Campylobacter coli
Authors: Roop R M , Smibert R M , Krieg N R
Published: January 2020

Campylobacter jejuni (20 strains) and Campylobacter coli (12 strains) were assigned to four biovars for each species based on phenotypic tests that were easy to perform and interpret. The resulting biotyping schemes offer a greater degree of distinction among C. jejuni and C. coli strains than any of the other biotyping schemes previously described for these organisms.
Serotyping and serology studies of campylobacteriosis associated with consumption of raw milk
Authors: Vogt R L , Little A A , Patton C M , Barrett T J , Orciari L A
Published: January 2020

A community outbreak of 15 cases of gastroenteritis was traced to consumption of unpasteurized milk produced at one commercial dairy. Using two different testing schemes, we found that a Campylobacter jejuni isolate from an ill patient and an isolate from a sick cow were the same serotype. Bacteriological studies suggested that a single epidemic strain of Campylobacter jejuni caused this outbreak.
Isolation of Streptococcus pneumoniae type 3 from equine species
Authors: Benson C E , Sweeney C R
Published: January 2020

Streptococcus pneumoniae type 3 was isolated from seven tracheobronchial aspirates and one pleural tap of seven adult horses and one foal. There was no direct evidence in these horses that isolation of the pneumococcus was related to a specific disease syndrome. Presenting complaints included two horses with chronic cough, two horses with decreased exercise tolerance, one horse with exercise-induced pulmonary hemorrhage, and three horses with pneumonia. Antibiotic therapy resolved the primary clinical complaint. This is the first report of the isolation of S. pneumoniae type 3 from adult horses.