bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthetic peptide-based immunoassays for distinguishing between human T-cell lymphotropic virus type I and type II infections in seropositive individuals
Authors: Lal R B , Heneine W , Rudolph D L , Present W B , Hofhienz D , Hartley T M , Khabbaz R F , Kaplan J E
Published: January 2020

Until now, serologic tests that distinguish the closely related human T-cell lymphotropic virus types I (HTLV-I) and II (HTLV-II) infections have not been available. Synthetic peptide assays, employing peptides derived from the core and envelope proteins of HTLV-I and HTLV-II (SynthEIA and Select-HTLV tests), were evaluated for the ability to serologically discriminate HTLV-I and HTLV-II infections. Of 32 HTLV-I- and 57 HTLV-II-positive serum specimens from individuals whose infections were confirmed by polymerase chain reaction, the SynthEIA test categorized 29 (91%) as HTLV-I and 50 (88%) as HTLV-II, and 10 (11%) were nontypeable. In contrast, the Select-HTLV test categorized 32 (100%) as HTLV-I and 55 (96%) as HTLV-II, and 2 (2%) were nontypeable. The specificity of both the assays in seropositive serum specimens was 100% in that none of the specimens were incorrectly classified. Additional serum specimens obtained from clinically diseased patients from the United States (n = 8) and asymptomatic carriers and patients from Japan (an endemic population for HTLV-I; n = 40) were categorized as HTLV-I by at least one of the assays, while serum specimens from Guaymi Indians from Panama (an endemic population for HTLV-II; n = 13) were categorized as HTLV-II. Thus, peptide enzyme immunoassays appear to represent a simple technique employing chemically synthesized antigens for discrimination between antibodies of HTLV-I and HTLV-II.
Use of chemotaxonomy as an aid to differentiate among Capnocytophaga species, CDC group DF-3, and aerotolerant strains of Leptotrichia buccalis
Authors: Bernard K , Cooper C , Tessier S , Ewan E P
Published: January 2020

Four strains of fastidious gram-negative rods, thought to be Capnocytophaga species (formerly CDC group DF-1 or Bacteroides ochraceus) or CDC group DF-3 on the basis of conventional phenotypic criteria, were also analyzed for cellular fatty acid (CFA) composition. It was found that the CFA compositions of these strains were qualitatively incorrect for those taxa. Subsequently, it was determined that all four bacteria were in fact aerotolerant strains of Leptotrichia buccalis, based on biochemical reactions, CFA composition, and lactic acid as the major end product of glucose fermentation. It is recommended that, in addition to conventional cultural and biochemical criteria, all strains of Capnocytophaga or CDC group DF-3 should also be tested for metabolic end products of fermentation and CFA composition as essential adjuncts for identification.
Analysis of human rotavirus strains prevailing in Bangladesh in relation to nationwide floods brought by the 1988 monsoon
Authors: Ahmed M U , Urasawa S , Taniguchi K , Urasawa T , Kobayashi N , Wakasugi F , Islam A I , Sahikh H A
Published: January 2020

The virologic character of human rotavirus strains prevailing in Bangladesh was investigated in relation to the devastating nationwide floods brought by the 1988 monsoon. Human rotaviruses contained in stool specimens that were collected from inpatients with infantile and adult diarrhea in two hospitals in Mymensingh over a 13-month period (January 1988 to January 1989) and in one hospital in Dhaka over a 3-month period (February to April 1988) were examined for their subgroup, VP7 serotype, and RNA electropherotype. In concurrence with the spread of the flood (from the middle of August 1988), the number of infantile and adult diarrhea patients increased greatly. At the same time, the proportion of rotavirus-positive specimens in all diarrhea cases also increased remarkably, reaching 54 and 45% in September and October, respectively. An electrophoretic analysis of viral RNA revealed 17 distinct patterns of viral RNA (14 long and 3 short electropherotypes) and a considerable number of mixed electropherotypes, suggesting the simultaneous infection of some patients with more than two rotavirus strains. It was noteworthy that electropherotypes of rotavirus strains prevailing in the community changed considerably after the spreading of the flood and that the frequency of virus specimens showing mixed electropherotypes increased significantly during the flood period. These results suggest that sudden environmental change caused by the devastating floods seriously affected the epidemiology of rotavirus infections by increasing the opportunity of transmission of the virus and by reducing the resistance of the host to infection. In both pediatric and adult patient groups, serotypes 1 and 2 were the most frequent ones detected, followed by serotype 4.(ABSTRACT TRUNCATED AT 250 WORDS)
Tests for detecting degradation of gelatin: comparison of five methods
Authors: Pickett M J , Greenwood J R , Harvey S M
Published: January 2020

Five methods for detecting degradation of gelatin by bacteria were compared. These were liquefaction in nutrient broth, hydrolysis in nutrient agar, hydrolysis of charcoal gelatin strips, degradation of the gelatin on strips of photographic film, and alkalinization of gelatin agar. Degradation of photographic film is a rapid and convenient method but, like hydrolysis of gelatin in broth and in agar, may fail to detect weakly positive strains of bacteria. Alkalinization of gelatin in an agar medium is a convenient and sensitive method to detect degradation of gelatin, particularly by Pseudomonas fluorescens, but this method may not be applicable to some species.
Comparison of preservation methods for enterotoxigenic Escherichia coli producing heat-labile enterotoxin
Authors: Yoh M , Narita I , Honda T , Miwatani T , Nishibuchi M
Published: January 2020

Ten strains of enterotoxigenic Escherichia coli producing heat-labile enterotoxin (LT) were preserved under 12 different conditions. After 1 month, 9 months, and 3 years of preservation, the cultures were recovered and examined for LT production. Preservation of the cultures on Dorset Egg Medium at 4 degrees C and preservation by freezing the cell suspensions in tryptic soy broth with 20% glycerol were found to be suitable preservation methods; all strains were alive for 3 years and had a minimum loss of LT production.
Vancomycin susceptibility and identification of motile enterococci
Authors: Vincent S , Knight R G , Green M , Sahm D F , Shlaes D M
Published: January 2020

Thirty-seven clinical isolates of Enterococcus gallinarum and Enterococcus casseliflavus and three type or reference strains of the species were studied with respect to vancomycin susceptibility and key identification characteristics. With the exception of one clinical isolate of E. casseliflavus (MIC, 4 micrograms/ml), MICs of vancomycin were 8 to 32 micrograms/ml. The type strain of E. gallinarum, NCDO 2313, and five of the clinical isolates had similar penicillin-binding protein profiles and shared 90 to 100% DNA homology. Two isolates, identified as E. gallinarum by conventional tests, were shown to be non-pigment-producing E. casseliflavus on the basis of penicillin-binding protein profile and DNA homology. The type and reference strains of E. casseliflavus, ATCC 25788 and ATCC 25789, were nonmotile in our experiments. However, both shared 65 to 100% DNA homology with each other and with five clinical isolates of E. casseliflavus. These data suggest that the MICs of vancomycin observed for strains of E. gallinarum and E. casseliflavus are higher than those usually associated with other enterococci and may be a common property of these species. Additionally, pigment production and motility may occasionally be misleading criteria for definitive identification of these organisms.
Detection and differentiation of antibodies to human T-cell lymphotropic virus types I and II by the immunofluorescence method
Authors: Gallo D , Penning L M , Hanson C V
Published: January 2020

We compared the sensitivities of the prototype human T-cell lymphotropic virus type I (HTLV-I)- and HTLV-II-transformed cell lines, MT2 and Mo-T, with that of an HTLV-II-infected cell line, clone 19, established in our laboratory, in the immunofluorescence (IF) test for detection of antibody to HTLV-I and HTLV-II. In addition, IF antibody titers with the three antigens were determined, and the results were compared with HTLV-I and HTLV-II typing by polymerase chain reaction (PCR). The MT2 cell line was more sensitive than the two HTLV-II cell lines for detecting HTLV-I antibody by IF, and clone 19 was more sensitive than Mo-T or MT2 for measuring HTLV-II antibody. In the titration study, the antigen that gave the highest titer correlated completely with the HTLV type determined by PCR, indicating that the relatively simple IF titration method can be used for differentiating HTLV-I and HTLV-II antibody in sera and plasmas.
Chemical characterization of clinical isolates which are similar to CDC group DF-3 bacteria
Authors: Daneshvar M I , Hollis D G , Moss C W
Published: January 2020

Six clinical isolates, taken from blood or wounds, that had biochemical characteristics most similar to Centers for Disease Control group DF-3 bacteria were examined for cellular fatty acid composition and isoprenoid quinone content to evaluate their chemical relatedness to known bacterial species and groups. The fatty acids were liberated from whole cells by base hydrolysis, methylated, and analyzed by capillary gas-liquid chromatography. The isoprenoid quinones were extracted from lyophilized whole cells and analyzed by reverse-phase high-performance liquid chromatography. All six strains, which were designated group DF-3-like, possessed a distinct fatty acid profile that was characterized by large amounts (greater than 20%) of 13-methyltetradecanoate (i-C15:0) and 12-methyltetradecanoate (a-C15:0), moderate amounts of saturated branched-chain 13-carbon acids (i-C13:0 and a-C13:0) and hexadecanoate (n-C16:0), and small to moderate amounts of both branched- and straight-chain hydroxy acids (i-3-OH-C15:0, 3-OH-C16:0, i-3-OH-C17:0, and 2-OH-C17:0). This fatty acid profile was unique compared with the profiles of group DF-3 and other bacteria we have previously tested and is useful for the rapid identification of group DF-3-like isolates. The isoprenoid quinone content of four group DF-3-like strains was similar, with ubiquinone-9 (Q-9) and Q-10 as their major quinones, while the other two group DF-3-like strains contained Q-7 as their major quinones, with smaller amounts of Q-8 and Q-9.
Use of Rambach Propylene Glycol Containing Agar for identification of Salmonella spp
Authors: Gruenewald R , Henderson R W , Yappow S
Published: January 2020

When grown on Rambach Propylene Glycol Containing Agar (Rambach agar), 216 of 230 (93.9%) Salmonella organisms isolated from patients and 54 of 62 (87.1%) Salmonella stock cultures produced a crimson-colored growth. Of the 14 clinical Salmonella isolates which displayed colors other than crimson, 8 were Salmonella typhi, 2 were Salmonella paratyphi A, and 4 belonged to other commonly isolated serotypes. All eight Salmonella stock cultures which failed to produce a crimson color belonged to rarely isolated serotypes. In contrast, of 83 non-Salmonella stock cultures distributed among 29 bacterial species, none produced a crimson color. These results suggest that while Rambach agar cannot preidentify S. typhi and S. paratyphi A, the medium can be used for the presumptive identification and can assist in the definitive identification of the overwhelming majority of Salmonella isolates.
Detection of Escherichia coli heat-stable enterotoxin genes in pig stool specimens by an immobilized, colorimetric, nested polymerase chain reaction
Authors: Hornes E , Wasteson Y , Olsvik O
Published: January 2020

A combination of selective enrichment by using immunomagnetic separation of F4 (K88)-positive Escherichia coli and a nested colorimetric polymerase chain reaction (PCR) was used on crude clinical and spiked samples for determination of genes encoding heat-stable enterotoxins (STs) Ia (ST Ia) and Ib (ST Ib). The combination increased the sensitivity of the nested PCR compared with that of application onto crude samples. Dead cells were also enriched by use of this technology, giving results that are not available by traditional cultivation as enrichment before PCR. The second step in the PCR was modified to be able to differentiate between ST Ia and ST Ib genes. The colorimetric PCR was performed in a microtiter format, making it useful for automation in clinical laboratories and for the screening of large numbers of samples.
Evaluation of the microbiology of chronic ethmoid sinusitis
Authors: Doyle P W , Woodham J D
Published: January 2020

In a prospective study, patients with the diagnosis of chronic ethmoid sinusitis were evaluated microbiologically by using biopsy specimens of the ethmoid sinus mucosa. Microbiology cultures were performed on 94 specimens from 59 patients. Staphylococcus aureus and members of the family Enterobacteriaceae were the most frequent classical pathogenic bacteria isolated. Coagulase-negative staphylococci were the most common overall isolates. Streptococcus pneumoniae and Haemophilus influenzae were infrequent isolates. No anaerobes, viruses, or Chlamydia trachomatis organisms were identified. Results of this study showed organism isolation frequencies different from those found in other studies of chronic sinusitis reported in the literature. The predominance of S. aureus and members of the family Enterobacteriaceae could have an effect on the antimicrobial therapy for chronic ethmoid sinusitis.
Clinical and laboratory features of Nocardia nova
Authors: Wallace R J , Brown B A , Tsukamura M , Brown J M , Onyi G O
Published: January 2020

Recent studies have shown that Nocardia asteroides isolates have five major antibiotic resistance patterns; one of these patterns identifies isolates of Nocardia farcinica. In the current study, we investigated a second pattern characterized by susceptibility to ampicillin and erythromycin. This pattern was seen in 17% of 223 clinical isolates identified by standard techniques as N. asteroides and associated with diseases typical for nocardiae. Biochemically, isolates with this drug pattern were relatively homogeneous and identical to the type strain and previous descriptions of Nocardia nova. The strains studied were unique among nocardiae in having both alpha- and beta-esterase activity (85 and 95%, respectively). However, the arylsulfatase activity at 14 days (75%) and antimicrobial susceptibility patterns, including susceptibility to erythromycin (100%), were the only routinely available methods that would separate N. nova strains from other members of N. asteroides. N. asteroides should be considered a complex because current clinical identification schemes include isolates of N. farcinica and N. nova and may well include additional species. This is the first detailed description of N. nova as a pathogen in humans.
Evaluation of infection with human immunodeficiency virus type 1 by using nonisotopic solution hybridization for detection of polymerase chain reaction-amplified proviral DNA
Authors: Coutlée F , Saint-Antoine P , Olivier C , Voyer H , Kessous-Elbaz A , Berrada F , Bégin P , Giroux L , Viscidi R
Published: January 2020

A convenient assay combining solution hybridization and enzyme immunoassay for DNA-RNA hybrids (polymerase chain reaction-enzyme immunoassay [PCR-EIA]) was developed to detect human immunodeficiency virus type 1 (HIV-1) provirus amplified by the PCR and was compared with oligomer hybridization with 32P-labeled SK19. In PCR-EIA, a fragment of the HIV-1 gag gene from peripheral blood mononuclear cells was first amplified with primer pair SK38/SK39 or O1/O2. PCR-amplified material was reacted in solution with a biotinylated RNA probe. Biotinylated hybrids were measured in a microtiter-plate EIA with antibiotin antibody and a beta-D-galactosidase-conjugated monoclonal antibody to DNA-RNA hybrids. Ten copies of HIV-1 DNA could be detected by PCR-EIA by using two different sets of primers. HIV-1 DNA was detected in 104 of 108 peripheral blood mononuclear cell samples by using SK38/39 and oligomer hybridization, in 104 of 108 samples by using SK38/SK39 and PCR-EIA, and in 104 of 108 samples by using O1/O2 and PCR-EIA. HIV-1 provirus was detected in 107 of 108 samples by using a combination of two sets of primers. One sample from a seropositive patient was negative in all three PCR assays, and six samples gave discordant results between primer pairs. Six of the latter samples scored negative in a PCR for beta-globin but became positive when the sample was diluted before amplification. When applied to clinical samples, PCR-EIA generated results similar to those of an isotopic assay for detection of amplified DNA.
Evaluation of acridinium-ester-labeled DNA probes for identification of Mycobacterium tuberculosis and Mycobacterium avium-Mycobacterium intracellulare complex in culture
Authors: Goto M , Oka S , Okuzumi K , Kimura S , Shimada K
Published: January 2020

The detectability of mycobacteria in culture by the use of nonisotopic, chemiluminescent DNA probes for Mycobacterium tuberculosis and the M. avium-M. intracellulare complex (MAC) was evaluated and compared with that by the use of 125I-labeled DNA probes for the same mycobacteria. In the assay, rRNA-directed DNA probes labeled with acridinium ester (AE-DNA probes) were used. Unhybridized probes were chemically degraded, and the esterified acridinium on the hybridized probes was hydrolyzed by the addition of alkaline hydrogen peroxide solution, resulting in the production of visible light which was measured with a luminometer. The detection limits of the AE-DNA probes were almost the same as those of the 125I-labeled DNA probes. A total of 107 clinical isolates of mycobacteria (47 isolates of M. tuberculosis, 36 MAC, and 24 atypical mycobacteria other than MAC) were tested. The sensitivity and specificity of the AE-DNA probes for M. tuberculosis were 100% both for the conventional method and with the 125I-labeled DNA probe. The sensitivity and specificity of the AE-DNA probes for MAC were 97.2 and 100%, respectively, for the conventional method and were both 100% with the 125I-labeled DNA probes. Because the procedure is simple, reliable, rapid (it can be completed within an hour), and safe (it does not use radioisotopes), it can easily be performed in any clinical laboratory.
Sensitivity of Limulus amebocyte lysate (LAL) to LAL-reactive glucans
Authors: Roslansky P F , Novitsky T J
Published: January 2020

The sensitivity of Limulus amebocyte lysate (LAL) to LAL-reactive glucans (LRGs) and lipid A was tested by using commercially available and experimentally formulated LAL reagents. The glucans included two kinds of beta-(1,3)-D-glucans, laminarin and curdlan, and cellulosic material, LAL-reactive material (LAL-RM), extracted from a hollow-fiber (Cuprophan) hemodialyzer. LAL-RM loses its LAL activity when it is digested with cellulase and thus appears to be a beta-(1,4)-D-glucan or a mixed glucan containing a substantial proportion of beta-(1,4) linkages. All LAL reagents tested were at least 1,000-fold more sensitive to endotoxin than to LRGs. The presence of the surfactant Zwittergent was shown to interfere with reactivity to LRGs; LAL reagents without added Zwittergent reacted more strongly to LRGs than did the same reagents containing Zwittergent. Chloroform extraction of LAL increased the reagents' sensitivity to both endotoxin and LRGs, but it was not responsible for LRG reactivity. The addition of Zwittergent significantly reduced the sensitivity of LAL reagents to lipid A. LAL without the surfactant was equally sensitive to endotoxin and lipid A. Both curdlan and LAL-RM amplified or enhanced the LAL response to endotoxin. Kinetic turbidimetric studies demonstrated that the enhancement was dependent on the glucan concentration.
Recovery of vancomycin-resistant gram-positive cocci from pediatric liver transplant recipients
Authors: Green M , Barbadora K , Michaels M
Published: January 2020

Between November 1988 and October 1989, 49 first-time pediatric liver transplant recipients at the Children's Hospital of Pittsburgh were prospectively monitored for the presence of stool colonization and the development of disease caused by vancomycin-resistant gram-positive cocci (VRGPC). Quantitative stool culturing was done on a weekly basis, and cultures were planted onto a selective medium for VRGPC. Isolates for which the MIC was greater than or equal to 8 were considered resistant to vancomycin. Patients were monitored clinically for the development of infection, and their charts were systematically reviewed for the use of antibiotics. Eighty-six isolates were recovered from 36 of the 49 patients. Enterococcal species were isolated from 31 patients and included Enterococcus gallinarum (n = 28), E. casseliflavus (n = 14), E. faecium (n = 9), E. faecalis (n = 2), E. mundtii (n = 2), and E. durans (n = 1). Stool colonization with vancomycin-resistant enterococci was noted to increase steadily during the first month after transplantation. Only 9 of 31 patients demonstrated clearance of these organisms in serial repeat cultures. Additional isolates of VRGPC included Lactobacillus confusus (n = 13), Lactobacillus spp. (n = 12), and Pediococcus pentosaceus (n = 4). Infection due to VRGPC developed in three patients: a urinary tract infection in two and peritonitis in one. E. faecium was the pathogen in each of these cases. The ranges of MICs of vancomycin were 8 to 32 micrograms/ml for all enterococcal isolates and greater than 128 micrograms/ml for Lactobacillus and Pediococcus isolates. All Lactobacillus and Pediococcus isolates were resistant to teicoplanin, although they were susceptible to daptomycin. All other isolates were susceptible to both teicoplanin and daptomycin. This study demonstrates that stool colonization with VRGPC may be a common and early finding among pediatric liver transplant recipients. However, infection appears to be uncommon.
Epidemiology of an endemic strain of beta-lactamase-producing Enterococcus faecalis
Authors: Patterson J E , Singh K V , Murray B E
Published: January 2020

Three previously reported beta-lactamase-producing (Bla+) enterococci with distinct but related antibiotic resistance phenotypes, plasmid profiles, and plasmid restriction endonuclease digestion patterns were isolated at the West Haven Veterans Administration Medical Center in Connecticut (WH245, WH257, WH571) in July 1986 and March 1987. In this study, we analyzed the whole-cell DNA of these isolates by using pulsed-field gel electrophoresis of large chromosomal fragments generated by SmaI digestion. The three West Haven isolates showed very similar chromosomal restriction endonuclease digestion patterns; these patterns were distinct from those of Bla+ and Bla- enterococci from other geographic areas and from those of other West Haven enterococci, suggesting an endemic Bla+ strain at this institution. Clinical information regarding these isolates suggests that exposure to the genitourinary clinic, obstructive urinary tract disease, and frequent antibiotic therapy may have been common risk factors for acquisition of this endemic strain.
Virulence factors associated with cytotoxic necrotizing factor type two in bovine diarrheic and septicemic strains of Escherichia coli
Authors: Oswald E , de Rycke J , Lintermans P , van Muylem K , Mainil J , Daube G , Pohl P
Published: January 2020

Forty-three bovine isolates of Escherichia coli producing a second type of cytotoxic necrotizing factor (CNF2) and three K-12 strains carrying different Vir plasmids coding for CNF2 were tested for the presence of several virulence factors. Most of the strains were serum resistant (79%), produced an aerobactin (70%), and adhered to calf villi (53%); some of them produced a colicin (32%) and a hemolysin (9%). These strains were also tested by a colony hybridization assay with gene probes for six toxins (classical heat-stable [STaP and STb] and heat-labile [LT-I and LT-IIa] enterotoxins and Shiga-like toxins [SLT-I and SLT-II]) and five adhesion factors (K99, K88, 987P, F17, and F41). Only two gene probes, LT-IIa (9%) and F17A (53%), hybridized with the CNF2 strains. However, antibodies raised against F17 fimbriae did not agglutinate the strains hybridizing with the F17A probe. In contrast, all except one of these strains adhered to calf villi. Interestingly, these two properties, F17A positivity and adherence to calf villi, were the only ones expressed by the K-12 strains carrying different Vir plasmids. In conclusion, this study confirmed that CNF2-producing strains are unrelated to previously described toxigenic E. coli strains and also demonstrated that in half of the strains the production of CNF2 was associated with an adhesion factor genetically related to, but different from, F17, which is more than likely encoded by Vir plasmids.
Use of a signature nucleotide sequence of hepatitis C virus for detection of viral RNA in human serum and plasma
Authors: Cha T A , Kolberg J , Irvine B , Stempien M , Beall E , Yano M , Choo Q L , Houghton M , Kuo G , Han J H
Published: January 2020

The nucleic acid sequence of the putative 5'-untranslated (5PUT) region of hepatitis C virus (HCV), determined for samples obtained from a variety of geographic origins, was found to be over 98% conserved among all isolates. On the basis of this signature sequence for HCV, a viral RNA assay was developed by using cDNA synthesis with reverse transcriptase, followed by polymerase chain reaction (PCR). The new assay was compared with the Ortho-Chiron C100-3 HCV enzyme-linked immunosorbent assay to research radioimmunoassays for antibodies to the C33c and C22 HCV antigens and to the first reported set of HCV PCR primers designed from the NS3 domain. Plasma samples from 16 Japanese patients with non-A, non-B hepatitis (NANBH) and 16 immunoassay-positive blood donors from the United States were investigated. The 5PUT PCR primers were found to be superior to the NS3 primers in sensitivity and specificity (15 of 25 versus 3 of 25 of the C100 enzyme-linked immunosorbent assay-positive samples, respectively). Samples from two C100-negative patients with acute NANBH were found to react with the 5PUT primers but not with the NS3 primers. Also, two of three patients with chronic NANBH converted from reverse transcriptase PCR positive to negative after interferon treatment. Although the clinical significance of the presence or absence of HCV RNA in samples from patients is not fully understood, the use of probes and primers from the 5PUT region (as opposed to primers from other segments) should not lead to false-negative results due to nucleic acid sequence variations in viral isolates.
Actinobacillus spp. and related bacteria in infected wounds of humans bitten by horses and sheep
Authors: Peel M M , Hornidge K A , Luppino M , Stacpoole A M , Weaver R E
Published: January 2020

We describe the isolation of Actinobacillus lignieresii and an A. equuli-like bacterium from an infected horse-bite wound in a 22-year-old stable foreman and A. suis from a bite injury in a 35-year-old man who had been attacked by a horse. A. lignieresii was also isolated in pure culture from an infected sheep-bite wound in a rural worker. These species of the genus Actinobacillus are primarily associated with animals and animal diseases and are rarely isolated from humans. The purpose of this report is to raise awareness of the possible occurrence of Actinobacillus spp. in bite wounds inflicted by farm animals and to discuss the difficulties encountered in the identification of species of Actinobacillus and related bacteria.
Direct polymerase chain reaction test for detection of Helicobacter pylori in humans and animals
Authors: Ho S A , Hoyle J A , Lewis F A , Secker A D , Cross D , Mapstone N P , Dixon M F , Wyatt J I , Tompkins D S , Taylor G R
Published: January 2020

We designed a polymerase chain reaction (PCR) for amplifying the Helicobacter pylori gene encoding 16S rRNA. Primers for the specific detection of H. pylori were designed for areas of the 16S rRNA gene in which there is the least sequence homology between H. pylori and its closest relatives. The specificity of detection was confirmed by ensuring that the primers did not amplify DNA extracts from the campylobacters H. cinaedi, H. mustelae, and Wolinella succinogenes, which are the closest relatives of H. pylori, as determined by 16S rRNA sequencing. Serial dilution experiments revealed the detection of as little as 0.1 pg of DNA by PCR and 0.01 pg by nested PCR. H. pylori DNA was detected successfully in clinical paraffin-embedded and fresh gastric biopsy specimens from patients positive for the bacterium and also in fecal suspensions seeded with the organism. The DNA from the nonculturable coccoid form of H. pylori was also identified by the primers. Universal primers designed for highly conserved areas on the 16S rRNA gene enabled large amplification products to be produced for direct sequencing analysis. Gastric bacteria resembling H. pylori have been isolated from animals. DNA of these animal gastric bacteria amplified with H. pylori-specific primers yielded PCR products identical to those from human isolates of H. pylori, as confirmed by the use of a 20-base radiolabelled probe complementary to an internal sequence flanked by the H. pylori-specific primers. The results of PCR amplification and partial 16S rRNA gene sequence analysis strongly support the contention that the gastric organisms previously recovered from a pig, a baboon, and rhesus monkeys are H. pylori.
Monoclonal antibody against Porphyromonas (Bacteroides) endodontalis lipopolysaccharide and application of the antibody for direct identification of the species
Authors: Hanazawa S , Sagiya T , Kitami H , Ohta K , Nishikawa H , Kitano S
Published: January 2020

The aim of the present study was to develop a monoclonal antibody that recognizes the shared antigen of Porphyromonas endodontalis so that we could use the antibody in direct identification and detection of P. endodontalis in infectious material from apical periodontal patients. We established a hybridoma cell line producing monoclonal antibody (BEB5) specific for P. endodontalis. BEB5 antibody reacted with all of the P. endodontalis strains tested, but not with any of the other black-pigmented Porphyromonas and Bacteroides spp. The antibody reacted specifically with the lipopolysaccharide (LPS) of three P. endodontalis strains of different serotypes (O1K1, O1K2, and O1K-). Western blotting (immunoblotting) analysis confirmed the specificity of the antibody to these LPSs, because the antibody recognized the typical "repetitive ladder" pattern characteristic of LPS on sodium dodecyl sulfate-polyacrylamide electrophoretic gels. These observations demonstrate that P. endodontalis LPS is the shared antigen of this species. The antibody can specifically identify P. endodontalis on nitrocellulose membrane blots of bacterial colonies grown on agar. The antibody is also capable of directly detecting the presence of P. endodontalis in infectious material by immunoslot blot assay. These results indicate that LPS is the shared antigen of P. endodontalis and that BEB5 antibody against LPS is a useful one for direct identification and detection of the organisms in samples from apical periodontal patients.
Respiratory syncytial virus infection and prevalence of subgroups A and B in Hawaii
Authors: Reese P E , Marchette N J
Published: January 2020

Respiratory syncytial virus was isolated from hospitalized children in Hawaii in each month of the year during the period January 1987 to August 1989. Subgroup A and subgroup B strains cocirculated, with subgroup A predominating. There was an alternating early-season and late-season peak incidence cycle as reported elsewhere.
Isoenzyme changes in Candida albicans during domestication
Authors: Lehmann P F , Wu L C , Mackenzie D W
Published: January 2020

Isoenzyme analysis was performed on multiple strains of two commonly used reference cultures of Candida albicans (B311 and NCPF3153). Whereas strains originating from C. albicans B311 showed no variation in isoenzyme profiles, some strains derived from NCPF3153 were identical to B311 strains but others showed variation in their glucose-6-phosphate dehydrogenase isoenzymes. The results are compared with those from previous analyses with these strains and show that C. albicans can undergo genetic alterations during prolonged maintenance in laboratories.
Specific amplification of Rickettsia tsutsugamushi DNA from clinical specimens by polymerase chain reaction
Authors: Furuya Y , Yoshida Y , Katayama T , Kawamori F , Yamamoto S , Ohashi N , Tamura A , Kawamura A
Published: January 2020

Polymerase chain reaction (PCR) was used to detect Rickettsia tsutsugamushi-specific DNA in clinical specimens. The primer pair used for PCR was designed from the nucleotide sequence of the gene encoding the 56-kDa antigen of the Gilliam strain. Theses primers led to a 78-bp fragment by amplifying the genomic DNAs from five serovariants, i.e., the Gilliam, Karp, Kato, Kawasaki, and Kuroki strains of R. tsutsugamushi, and also the DNA from blood clots of patients with scrub typhus, even at the early stage of onset of the disease. This indicates that this method is suitable for the diagnosis of scrub typhus.
Gas-liquid chromatographic analysis of cellular fatty acids for identification of gram-negative anaerobic bacilli
Authors: Stoakes L , Kelly T , Schieven B , Harley D , Ramos M , Lannigan R , Groves D , Hussain Z
Published: January 2020

A commercially available, computer-assisted microbial identification system (MIS) employs gas-liquid chromatographic analysis of cellular fatty acids for bacterial identification. MIS was compared with conventional identification systems. Of 225 gram-negative anaerobes tested, MIS identified 72.4% of the strains to the species level, 88.9% to the appropriate group, and 93.3% to the correct genus.
Cytomegalovirus detection in transplant patients: comparison of different methods in a prospective survey
Authors: Wunderli W , Auracher J D , Zbinden R
Published: January 2020

In a prospective survey, all transplant patients at the hospital of the University of Zurich were screened for cytomegalovirus (CMV) infection. CMV infections were detected in a total of 40 of 104 transplant recipients; 31 could be diagnosed by CMV immunoglobulin M serology, 27 could be diagnosed by viremia, 11 could be diagnosed by antigenemia, and 13 could be diagnosed by the start of virus secretion. Combined application of serology and the detection of viremia showed the highest sensitivity (39 of 40 cases). Of the patients with severe clinical symptoms, six of seven had primary CMV infections caused by a positive transplant. Therefore, it is strongly indicated that patients with known risk factors should be regularly surveyed by a combination of methods.
Failure of Micro Media FOX Microdilution System to detect decreased susceptibility of Streptococcus pneumoniae to penicillin
Authors: Carroll K , Cohen S , Saxon B , Hamilton L
Published: January 2020

Twelve isolates of Streptococcus pneumoniae with decreased susceptibility by oxacillin screen were susceptible by the FOX panel (Micro Media Systems, Cleveland, Ohio), a commercial microdilution system designed for fastidious organisms. These organisms were found to be moderately susceptible or resistant by broth macrodilution and agar dilution methods. This discrepancy indicates that the FOX panel is not reliable for susceptibility testing of S. pneumoniae.
Rapid diagnosis of respiratory syncytial virus infection by using Pernasal swabs
Authors: Mackie P L , Madge P J , Getty S , Paton J Y
Published: January 2020

The sensitivity and specificity of direct immunofluorescence microscopy performed on Pernasal swab specimens and compared with those of nasopharyngeal aspirates were 93 and 99%, respectively. Posterior nasopharyngeal swabs applied immediately to microscope slides allow a rapid and simple screening procedure for the diagnosis of acute respiratory syncytial virus infections.
Immunodiffusion test for diagnosis and monitoring of human pythiosis insidiosi
Authors: Pracharktam R , Changtrakool P , Sathapatayavongs B , Jayanetra P , Ajello L
Published: January 2020

To facilitate the laboratory diagnosis of human cases of pythiosis insidiosi, an immunological test was evaluated. A soluble antigen was prepared from a human isolate of Pythium insidiosum, an aquatic, thermotolerant oomycete that causes infections in cattle, dogs, horses, and humans. Sera from seven proven cases of disseminated human pythiosis insidiosi were tested in an immunodiffusion test along with appropriate control sera from patients with a variety of actinomycotic, bacterial, and mycotic diseases as well as sera from uninfected individuals. Titers ranged from 1:1 to 1:32 in the seven serum samples from the disseminated cases of pythiosis insidiosi of varying severity. The heterologous sera gave negative reactions. The rapidity and specificity of the immunodiffusion test makes it a useful diagnostic tool for the serodiagnosis of P. insidiosum infections.
Ampicillin-resistant Enterococcus raffinosus in an acute-care hospital: case-control study and antimicrobial susceptibilities
Authors: Chirurgi V A , Oster S E , Goldberg A A , Zervos M J , McCabe R E
Published: January 2020

A prospective study identified 9 (32%) of 28 ampicillin-resistant (MIC greater than or equal to 16 micrograms/ml) enterococcus isolates as Enterococcus raffinosus. A case-control study found no significant differences with respect to underlying diseases, catheterization, or surgery between patients with ampicillin-resistant E. raffinosus and those with ampicillin-susceptible Enterococcus spp. Prior treatment with antibiotics and prolonged hospitalization were more frequent among patients with ampicillin-resistant E. raffinosus. Patients with the same strain (determined by plasmid analysis) were frequently hospitalized concurrently.
Homotypic and heterotypic serological responses to rotavirus neutralization epitopes in immunologically naive and experienced animals
Authors: Snodgrass D R , Fitzgerald T A , Campbell I , Browning G F , Scott F M , Hoshino Y , Davies R C
Published: January 2020

Gnotobiotic or specific-pathogen-free animals with no previous exposure to rotavirus were vaccinated with strain UK, serotype G6. The highest serological response was to homologous virus; significant but lower responses occurred to viruses with either VP4 or VP7 related to that of vaccine virus; responses to other viruses were of low titer or infrequent. Adult cows vaccinated with UK virus produced increased titers of antibody to all rotavirus serotypes. The increases in titer to homologous virus and to other natural and reassortant viruses sharing VP7 with the vaccine virus were significantly higher than those to all other viruses. These results suggest the presence of common epitopes which are not well recognized in primary infections.
Mycobacterium malmoense: an easily missed pathogen
Authors: Hoffner S E , Henriques B , Petrini B , Källenius G
Published: January 2020

The BACTEC system (Becton Dickinson, Sparks, Md.) was compared with culture on Löwenstein-Jensen egg medium for the detection of Mycobacterium malmoense in lymph node samples from children with lymphadenitis. It was found to be significantly more sensitive and rapid. The use of the BACTEC system is thus recommended as a complement to culture on solid media for the primary isolation of atypical mycobacteria, such as M. malmoense.
Enteroinvasive Escherichia coli: a cause of bacteremia in patients with AIDS
Authors: Bessesen M T , Wang E , Echeverria P , Blaser M J
Published: January 2020

A strain of enteroinvasive Escherichia coli was isolated from the blood of a patient with advanced human immunodeficiency virus disease on repeated occasions, associated with severe diarrheal illness. The isolate was killed in vitro by control sera but not by sera collected from the patient before or after his bacterial illnesses.
Arm abscesses caused by Clostridium botulinum
Authors: Elston H R , Wang M , Loo L K
Published: January 2020

Wound botulism is an uncommon disorder that continues to be rarely reported in the United States. A 34-year-old intravenous heroin user was admitted to the Loma Linda, Calif., Veterans Administration hospital with multiple abscesses on his forearms. His clinical course was compatible with botulism, and his culture and serum were positive for Clostridium botulinum toxin type A. Early culture and/or serum identification can lead to prompt diagnosis, treatment, and improvement in the morbidity and mortality rates of this disease.
Mycobacterium avium complex disease in patients with AIDS: seroreactivity to native and recombinant mycobacterial antigens
Authors: Morris S L , Bermudez L , Chaparas S D
Published: January 2020

Antibodies to Mycobacterium avium complex (MAC) antigens were measured by enzyme-linked immunosorbent assays and immunoblot analyses in sera from 20 patients with AIDS and disseminated MAC disease, 5 human immunodeficiency virus-seronegative patients with pulmonary MAC infections, and 20 healthy controls. Whereas enzyme-linked immunosorbent assay titers for healthy controls and patients with AIDS and MAC disease were comparable, human immunodeficiency virus-seronegative patients with MAC disease had higher anti-MAC antibody titers (P less than 0.01). Immunoblot analysis with the same sonic extracts indicated that each of the three groups had a limited heterogeneous response to M. avium antigens. No significant differences in immunoblot reactivities were detected. However, immunoblot studies with recombinant nontuberculous mycobacterial antigens revealed that sera from over 90% of the patients with MAC disease and only 25% of controls recognized a recombinant protein derived from a 35-kDa mycobacterial antigen. Although sonic extracts did not permit adequate discrimination of antibody reactivity in patients with MAC disease, recombinant antigens may be useful as indicators of disease.
Differentiation of Lactococcus lactis and Lactococcus garvieae from humans by comparison of whole-cell protein patterns
Authors: Elliott J A , Collins M D , Pigott N E , Facklam R R
Published: January 2020

We tested 12 reference and 24 clinical strains of lactococci for physiologic characteristics using a conventional test system, the Gen-Probe Enterococcus 2 chemiluminescence assay (Gen-Probe Inc., San Diego, Calif.), the Rapid Strep identification system (Analytab Products, Plainview, N.Y.), and whole-cell protein analysis. The Gen-Probe Enterococcus 2 chemiluminescence assay for Enterococcus identification was negative with all strains. Neither the conventional test nor the Rapid Strep identification system could differentiate between the two Lactococcus spp. most commonly isolated from humans. A simple procedure, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was developed for comparing the whole-cell protein patterns of Lactococcus spp. L. lactis and L. garvieae were differentiated by unique protein patterns.
Ability of National Committee for Clinical Laboratory Standards-recommended quality control strains from the American Type Culture Collection to detect errors in disk diffusion susceptibility tests
Authors: Yechouron A , Dascal A , Stevenson J , Mendelson J
Published: January 2020

The National Committee for Clinical Laboratory Standards (NCCLS) recommends, as a quality control for the disk diffusion susceptibility test, the use of three strains from the American Type Culture Collection: Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, and Escherichia coli ATCC 25922. This study assesses the capacity of these strains to detect errors in the overall method. ATCC strains were tested by comparing testing by the standard NCCLS-recommended procedure (ST) with testing under the following conditions: incubation at 25 degrees C, Mueller-Hinton agar depths of 2 mm (AD2) and 8 mm (AD8), agar pHs of 6.5 and 8, inocula with McFarland standards of 0.25 (0.25M) and 4.0 McFarland (4.0M), direct inoculation without preincubation of inoculum (DI), and a 2-h delay between inoculation and disk application (2HR). The frequency of zone measurements outside the NCCLS-recommended control zone limits were as follows: ST, 0%; AD2, 18%; AD8, 9.6%; pH 6.5, 7.9%; pH 8, 5.3%; 0.25M, 3.5%; 4.0M, 24%; DI, 3.4%; 2HR, 1.8%; 25 degrees C (only E. coli and P. aeruginosa were evaluable), 28%. These results suggest that the quality control strains are only partially effective in detecting single extreme laboratory errors and that careful laboratory supervision is necessary even in the setting of properly monitored quality control strains.
Comparison of the Vitek Immunodiagnostic Assay System with an indirect immunoassay (Toxostat Test Kit) for detection of immunoglobulin G antibodies to Toxoplasma gondii in clinical specimens
Authors: Sandin R L , Knapp C C , Hall G S , Washington J A , Rutherford I
Published: January 2020

The value and convenience of testing for specific anti-Toxoplasma gondii antibodies have led to the development of various antibody detection methods, such as the enzyme immunoassay. Two enzyme immunoassays, the Vitek Immuno-Diagnostic Assay System (VIDAS; Vitek Systems, Hazelwood, Mo.) and the Toxostat Test Kit (TST; Whittaker Bioproducts, Walkersville, Md.), were compared for their ability to detect T. gondii immunoglobulin G antibodies in fresh human sera. Specimens were tested according to the instructions of each manufacturer. Of 304 serum specimens tested, the results for 282 (93%) agreed in both assays (77 were positive and 205 were negative). The results for the remaining 22 (7%) specimens were discrepant; 20 of the 22 specimens tested low positive or equivocal by Toxostat and negative by VIDAS. Upon retesting, 8 of these 22 specimens were in concordance, increasing the level of agreement to 95.3%. Intra-assay reproducibility was tested with four to nine replicates of each of seven samples (four positive, one negative, and two equivocal) by both tests and six replicates of one standard by VIDAS. Coefficients of variation (CVs) for VIDAS were 6, 8, 10, 15, 18, 19, and 23% for the samples and 14% for the standard. For Toxostat the CVs were 2, 8, 10, 10, 13, and 16%. In general, CVs for VIDAS were slightly larger than the CVs for Toxostat. Day-to-day variability over 3 days was tested by VIDAS for only six samples (three positive, two negative, and one equivocal) and four standards; CVs ranged from 2 to 15% for the samples and from 6 to 13% for the standards. In conclusions, VIDAS is a rapid, convenient, non-labor-intensive, and easily performed test for the detection of immunoglobulin G antibodies to T. gondii in serum specimens.
Molecular relatedness of Staphylococcus epidermidis isolates obtained during a platelet transfusion-associated episode of sepsis
Authors: Shayegani M , Parsons L M , Waring A L , Donhowe J , Goering R , Archinal W A , Linden J
Published: January 2020

Staphylococcus epidermidis was isolated from the blood of a 25-year-old pregnant woman following the administration of eight units of platelets. She had developed chills and a fever of 41.4 degrees C soon after the transfusions were completed. S. epidermidis was also obtained from all eight platelet units, as well as from the packed-erythrocyte unit associated with the first unit of platelets. The isolation of the same organism from these epidemiologically related sources provided us with the opportunity to phenotypically and genetically characterize the isolates. Several typing methods, including four molecular techniques, were used to increase our chances of finding any differences between the isolates under investigation. Phenotypic analyses demonstrated that S. epidermidis isolates from the patient, platelet units, and erythrocyte unit reacted in exactly the same manner in 15 biochemical tests, exhibited slime production, and had the same antibiotic susceptibility pattern. Genetic analyses, which included plasmid profiles, plasmid cross-hybridization, field inversion gel electrophoresis, and ribotyping, substantiated the relationships between the S. epidermidis isolates from the patient, platelet units, and erythrocyte unit. Eight S. epidermidis control strains unrelated to the case were found to differ significantly from the platelet-related strain.