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PubMed1986

Identification and its vicissitudes in the psychoses. The importance of the concept of the 'maddening object'.

This paper describes 'psychotizing bonds' in terms of identification processes, the way they function in the constitution of the psychic apparatus and their relation to the deficient self. The author relates the pathogenic potentiality of the psychotic nuclei with the tendency of psychotic disorganization to be irreversible. Psychotic regression is considered in terms of pathogenic identification with forms of ego and superego functioning that belong to the primitive parental objects of infancy. Whereas normogenic identifications structure the subject's own ego resources, the pathogenic identifications which appear in the psychotic transference, form bonds that stifle spontaneity and force the self to be transformed into the other. These ideas lead thus to the concept of the 'maddening object'. Finally, the pathogenic identifications in the psychoanalytic process are examined. The patient should be 'rescued' from these bonds linking the self with the maddening objects. The analyst must hold the conviction that a virtual and potential subject exists in the analysand, in spite of his psychotic condition. The deficient self which becomes manifest during the moments of dis-identification must be assisted.

Adult↗
PubMed2004

[Application of molecular biology techniques in the identification of pathogenic fungi and the diagnosis of fungal infection].

With the increasing incidence and mortality of fungal infection, the requirements for strict diagnostic approaches became a very urgent issue. Because of the traditional detective techniques, such as culture, gave poor diagnostic outcomes, the molecular biological techniques are expected to develop the potential diagnostic approaches. During the past decades, we have carried out serial studies on the molecular properties of pathogenic fungi, and we would like to review as following. Firstly, we applied several molecular tools in classification and identification of pathogenic fungi. We performed random amplification of polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) and other techniques in studying the typing, to classify and identify the properties of Dermatophytes, Candida spp., Cryptococcus neoformans, Dematiaceous fungi, and Aspergillus spp. Interestingly, we found the same T. rubrum strain might infect different sites of the host, while a site-specificity displayed in T. mentagrophytes. This finding indicated the genetic discrepancies among the fungi. Beside, we also found that the E. dermatitis strains with different virulences possessed some discrepancies at gene level. We then developed a PCR-based molecular procedure to identify the novel species in Exophiala spp. As the applicable strategy, we also investigated the rDNA sequence properties in several fungi. And as a result, we submitted for the first time to GenBank the complete sequence of Aspergillus fumigatus rDNA/ITSI/ITSII, which provided the basis for designing the species-specific probes and for its further clinical applications. Secondly, we have tried to develop the molecular diagnostic approaches based on our DNA sequence data which were used for identification studies previously. By analyzing the DNA sequence of Aspergillus fumigatus rDNA/ITSI/ITSII, we developed a nested PCR method to detect Aspergillus fumigatus genes. Our preliminary results indicated that this PCR-based molecular approach has great importance in the diagnosis of invasive aspergillosis. We also designed the species-specific probes and then established several in situ hybridization procedures. We found these hybridization methods could get the positive rate up to 81% (13/16), which suggests that these methods have potential diagnostic value for invasive candidiasis and aspergillosis. Based on our experiences, we would conclude that the molecular biological techniques possess great value to investigate the biological properties of pathogenic fungi, and we are looking forward to see more and more molecular tools will be used in the pathogenic mechanisms of fungal infections and antifungal activity studies.

Aspergillosis↗
PubMed1988

Evaluation of a ten-minute chromogenic substrate test for identification of pathogenic Neisseria species and Branhamella catarrhalis.

A ten-minute chromogenic substrate test was evaluated for its ability to rapidly identify pathogenic Neisseria spp. and Branhamella catarrhalis. Identifications obtained with this system were compared to those obtained using conventional procedures. The test correctly identified 98.9% of 90 Neisseria gonorrhoeae, 98.3% of 60 Neisseria meningitidis, 96.2% of 26 Neisseria lactamica, and 100% of 36 Branhamella catarrhalis strains. Eight Neisseria subflava strains that grew on modified Thayer-Martin agar were prolyl aminopeptidase positive and were misidentified as Neisseria gonorrhoeae. Other strains of saprophytic Neisseria spp. also reacted with the chromogenic substrates. The system was accurate and reliable for identifying the commonly encountered pathogenic species. In light of recent reports describing new species and atypical Neisseria strains, however, careful attention to the salient features of both common and atypical organisms is necessary for proper use of rapid enzymatic identification tests.

Chromogenic Compounds↗
PubMed1994

The use of lipid-linked oligosaccharides (neoglycolipids) in the identification of carbohydrate receptors for microbial pathogens.

Specific oligosaccharide chains on the host cell surface act as receptors for many microbial pathogens. Identification of receptor structures is an important step in the understanding of the pathogenesis of infection. Glycolipid receptors have been identified by direct binding assays. However, technical difficulties have prevented demonstration of bacterial binding to the oligosaccharides of glycoproteins; these have been identified mainly by inhibition assays. By a novel technique developed in our laboratory, oligosaccharides released from glycoproteins are linked to lipids to form neoglycolipids. These can be used in bacterial binding assays. The feasibility of this approach has been demonstrated using type 1 fimbriated Escherichia coli binding specifically to neoglycolipids rich in mannose residues. The application of the method has resulted in a demonstration of a new type of adhesive specificity for E. coli and differences in the binding specificities of E coli and Pseudomonas aeruginosa. Further application of this technique by generating oligosaccharides purified from mucus glycoproteins from patients with cystic fibrosis to use in binding assays with P aeruginosa is currently being undertaken. The basic knowledge gained by this approach may in time see the development of novel therapy in the form of receptor blocking agents.

Bacteria↗
PubMed2005

McRAPD as a new approach to rapid and accurate identification of pathogenic yeasts.

Despite advances in antifungal prophylaxis and therapy, morbidity and mortality incurred by yeasts remain a significant burden. As pathogenic yeast species vary in their susceptibilities to antifungal agents, clinical microbiology laboratories face an important challenge to identify them rapidly and accurately. Although a vast array of phenotyping and genotyping methods has been developed, these are either unable to cover the whole spectrum of potential yeast pathogens or can do this only in a rather costly or laborious way. Random amplified polymorphic DNA (RAPD) fingerprinting was repeatedly demonstrated to be a convenient tool for species identification in pathogenic yeasts. However, its wider acceptance has been limited mainly due to special expertise and software needed for analysis and comparison of the resulting banding patterns. Based on a pilot study, we demonstrate here that a simple and rapid melting curve analysis of RAPD products can provide data for identification of five of the most medically important Candida species. We have termed this new approach melting curve of random amplified polymorphic DNA (McRAPD) to emphasize its rapidity and potential for automation, highly desirable features for a routine laboratory test.

Candida↗
PubMed2005

Natural history of pulmonary complications in children after bone marrow transplantation.

We sought, in children after bone marrow transplantation (BMT), (1) to determine the natural history and incidence of pulmonary complications, (2) to evaluate the diagnostic yield of fiberoptic bronchoscopy and bronchoalveolar lavage (BAL); and (3) to determine the effect of bronchoscopy with lavage on patient outcome. The study design was a retrospective review in a tertiary care university hospital of all children undergoing BMT over a 5-year period. Patients were separated into 2 study groups: children with and without pulmonary complications. Pulmonary complications were defined as new or persistent pulmonary infiltrates on chest radiograph or chest computed tomography scan, respiratory symptoms, hypoxemia, or hemoptysis. Three hundred sixty-three pediatric patients underwent BMT between January 1, 1995, and December 31, 1999. Ninety patients (25%) developed pulmonary complications and were evaluated with bronchoscopy and BAL. Patients with pulmonary complications had a higher mortality (65% versus 44%; P < .01). The median posttransplantation survival for children with pulmonary complications was 258 days, compared with 1572 days in patients without pulmonary complications. The incidence of pulmonary complications was increased in patients with allogeneic BMT (P < .01). The time-dependent onset of severe (grade III to IV) graft-versus-host disease increased the relative risk of pulmonary complications by 2.0 (95% confidence interval, 1.1-3.7; P = .02). Pulmonary complications increased the time-dependent relative risk of mortality by 3.5 (95% confidence interval, 2.5-4.8). The diagnostic yield of bronchoscopy with lavage was 46% in patients undergoing BAL. Diagnostic bronchoscopy did not enhance either 30- or 100-day survival. Pathogen identification did not decrease mortality (P = .45). Pulmonary complications occur in 25% of children undergoing BMT and increase the risk of death in the first year after BMT. Although pathogen identification does not confer a survival advantage, rigorous, prospective screening may allow for earlier identification of pathogens and thereby provide a benefit to this uniquely vulnerable population.

Adolescent↗
PubMed2006

A microbial diagnostic microarray technique for the sensitive detection and identification of pathogenic bacteria in a background of nonpathogens.

A major challenge in microbial diagnostics is the parallel detection and identification of low-bundance pathogens within a complex microbial community. In addition, a high specificity providing robust, reliable identification at least at the species level is required. A microbial diagnostic microarray approach, using single nucleotide extension labeling with gyrB as the marker gene, was developed. We present a novel concept applying competitive oligonucleotide probes to improve the specificity of the assay. Our approach enabled the sensitive and specific detection of a broad range of pathogenic bacteria. The approach was tested with a set of 35 oligonucleotide probes targeting Escherichia coli, Shigella spp., Salmonella spp., Aeromonas hydrophila, Vibrio cholerae, Mycobacterium avium, Mycobacterium tuberculosis, Helicobacter pylori, Proteus mirabilis, Yersinia enterocolitica, and Campylobacter jejuni. The introduction of competitive oligonucleotides in the labeling reaction successfully suppressed cross-reaction by closely related sequences, significantly improving the performance of the assay. Environmental applicability was tested with environmental and veterinary samples harboring complex microbial communities. Detection sensitivity in the range of 0.1% has been demonstrated, far below the 5% detection limit of traditional microbial diagnostic microarrays.

Bacteria↗
PubMed1998

Streptococcus iniae, a human and animal pathogen: specific identification by the chaperonin 60 gene identification method.

It was recently reported that Streptococcus iniae, a bacterial pathogen of aquatic animals, can cause serious disease in humans. Using the chaperonin 60 (Cpn60) gene identification method with reverse checkerboard hybridization and chemiluminescent detection, we identified correctly each of 12 S. iniae samples among 34 aerobic gram-positive isolates from animal and clinical human sources.

Animals↗
PubMed1997

Development of a highly specific assay for rapid identification of pathogenic strains of Yersinia enterocolitica based on PCR amplification of the Yersinia heat-stable enterotoxin gene (yst).

The chromosomal gene yst, which encodes a heat-stable enterotoxin of Yersinia enterocolitica, is a useful diagnostic marker because it occurs only in invasive strains of this species. A homologous gene also occurs in some strains of Yersinia kristensenii. Sequence analysis of the yst genes from two different strains of Y. enterocolitica and from Y. kristensenii revealed a substantial number of mismatches at the 3' ends of the yst genes of the so-called American and European biotypes of Y. enterocolitica. Moreover, several mismatches and a deletion of 5 codons were found in the yst of Y. kristensenii. These findings were used to develop a PCR-based assay for yst of Y. enterocolitica which yielded a detectable product in as little as 50 min. The assay was 100% specific in terms of its ability to identify potentially pathogenic strains of Y. enterocolitica regardless of biotype or serotype. The PCR yielded an amplicon that was visible on agarose gel electrophoresis from as few as 100 CFU, or 10 CFU when the PCR was combined with dot blot hybridization with a digoxigenin-labeled oligonucleotide probe that corresponded to an internal sequence of yst. These results establish the value of the yst gene as a target for the identification of pathogenic bioserotypes of Y. enterocolitica and the usefulness of PCR for this purpose.

Amino Acid Sequence↗
PubMed1993

Digoxigenin-labelled inv- and ail-probes for the detection and identification of pathogenic Yersinia enterocolitica in clinical specimens and naturally contaminated pig samples.

A non-radioactive colony hybridization method was developed for the rapid detection of Yersinia enterocolitica in primary isolates and for differentiation between pathogenic and non-pathogenic strains. The method is based on, respectively, the presence of the inv-locus in all Yersinia spp. and the presence of the ail-gene in pathogenic Y. enterocolitica only. Hybridization results with ail-probes of 132 strains of Y. enterocolitica were in good agreement with pathogenicity phenotypes as indicated by a tissue culture invasion (TCI) assay and by serotyping. All TCI+ strains and only two TCI- strains were positive by hybridization with ail. Hybridization results with inv- or ail-probes of 150 primary isolates of human, animal or slaughterhouse origin were compared with those of conventional methods to detect and identify Y. enterocolitica. All samples that were positive for Yersinia spp. by cultivation (four of 66) or were positive for pathogenic Y. enterocolitica by cultivation and serotyping (six of 84) were also positive by hybridization with, respectively, the inv- or ail-probe. In three slaughterhouse swab samples, in which Yersinia spp. were not detected by cultivation (2%), strong positive hybridization signals were obtained with the inv- and/or ail-probe. Four other swab samples which were negative by cultivation produced weak positive signals by hybridization with inv- and/or ail-probes. These results indicate that the method can be used for (1) the identification of pathogenic Y. enterocolitica isolates and (2) the detection of Yersinia spp. in primary isolates of naturally contaminated samples.

Abattoirs↗
PubMed2004

Identification of pathogenic dematiaceous fungi and related taxa based on large subunit ribosomal DNA D1/D2 domain sequence analysis.

The nucleotide sequences of the D1/D2 domains of large subunit (26S) ribosomal DNA for 76 strains of 46 species of pathogenic dematiaceous fungi and related taxa were determined. Intra-species sequence diversity of medically important dematiaceous fungi including Phialophora verrucosa, Fonsecaea pedrosoi, Fonsecaea compacta, Cladophialophora carrionii, Cladophialophora bantiana, Exophiala dermatitidis, Exophiala jeanselmei, Exophiala spinifera, Exophiala moniliae, and Hortaea werneckii were extremely small; as few as 0 changes were detected in C. bantiana, Fonsecaea and Exophiala species, 1 bp in C. carrionii and H. werneckii, and 2 bp in P. verrucosa. Inter-species nucleotide diversity between most species was higher. These data suggested that the D1/D2 domain is sufficiently variable for identification of pathogenic dematiaceous fungi and relevant species. The phylogenetic trees constructed from the sequence data revealed that most human pathogenic species formed a single cluster and that Cladosporium and Phialophora species were distributed polyphyletically into several clusters.

Ascomycota↗
PubMed1994

Transfer RNA profiling: a new method for the identification of pathogenic Candida species.

A new molecular taxonomic method applicable to the identification of medically important Candida species and other yeast species has been developed. It is based on the electrophoretic pattern of total tRNA samples (a 'tRNA profile') isolated from Candida species and generated using high-resolution semi-denaturing urea-polyacrylamide gel electrophoresis and methylene blue staining. Species-specific tRNA profiles for the species C. albicans, C. tropicalis, C. parapsilosis, C. guilliermondii, C. glabrata and Pichia guilliermondii were obtained. Detailed studies with the major human pathogen of the Candida genus, C. albicans, demonstrated that the tRNA profile for a given species was both reproducible and strain-independent; seven different C. albicans strains generated identical tRNA profiles. Minor strain-specific heterogeneities in the tRNA profiles of C. guilliermondii and C. parapsilosis were detected, but in neither case did they significantly alter the species-specific diagnostic tRNA profile. The potential of this method in clarifying taxonomic anomalies was demonstrated by the finding that Type I and Type II strains of C. stellatoidea generate very different tRNA profiles, with that of a Type II strain being identical to the C. albicans tRNA profile. This method offers a number of advantages over current electrophoretic karyotype methods for species identification, both within the Candida genus and with yeast species in general.

Candida↗
PubMed2005

Effect of pathogen-specific clinical mastitis on herd life in two New York State dairy herds.

The objective of this study was to estimate the effects of clinical mastitis (CM) (both with and without specific pathogen identification) occurring in different stages of lactation on length of herd life in two New York State dairy farms. The 2,697 cows in the study were followed for one lactation (the first-occurring one on or after 1 October 1999), until it ended because of a new lactation, culling, or end of study (31 March 2001 in one farm; 31 July 2001 in the other). A Cox proportional hazards model with time-dependent covariates, in SAS((R)), was used to measure, within a lactation, the effect of the first occurrence of CM (without specific pathogen identification) occurring 1--7, 8--66, 67--100, 101--225, or >or=226 days in milk (DIM), on how long cows remained in the herd. For the first occurrence of CM due to Streptococcus spp., Staphylococcus aureus, Staphylococcus spp., Escherichia coli, Klebsiella spp., and 'no pathogen isolated', the intervals were before and after the median DIM of first occurrence of each pathogen. There were too few cases due to Arcanobacterium pyogenes, and 'other pathogens grouped together' to split into intervals, so they were modeled as binary variables, i.e. as they occurred. CM was modeled using time-dependent covariates, to account for its differing effects throughout lactation on culling. Other variables controlled for were herd, parity, calving season, and other significant diseases. In the dataset, the lactational incidence risk of the first occurrence of CM was 18.2%; 20.0% of the cows did not survive the lactation that was studied. The overall annual culling percentage for both herds during the study period (including all cows, whether eligible for the study or not) was 35.6%. For cows with CM without pathogen identification, their highest hazard ratio (HR) of culling occurred from 67 to 100 DIM. All of the pathogens modeled markedly reduced herd life. On average over the entire lactation, cows with Staphylococcus spp. CM had the highest HRs for culling, although there were no significant differences among pathogens (at p=0.0018 (reflecting 28 pairwise comparisons)). For early-occurring (before median DIM of first occurrence) S. aureus CM, the daily rate of change of the HR of culling increased over time. The HRs for culling were particularly high for late-occurring (after median DIM of first occurrence) E. coli and Klebsiella spp. CM early in the interval, but the daily rate of change of the hazard of culling for these two pathogens decreased sharply over time. Treating CM as time-dependent therefore allowed us to measure in greater detail, its varying effects (of when it occurred) on herd life.

Animals↗
PubMed1983

Identification of pathogenic Neisseria species with the RapID NH system.

The RapID NH system (Innovative Diagnostics Systems, Inc., Decatur, Ga.) is a 4-h test used in the identification of Neisseria and Haemophilus species. The system was evaluated for accuracy and reliability and compared with conventional (cystine proteose peptone agar; Prepared Media Laboratory, Tualatin, Ore.) carbohydrate degradation tests with Neisseria gonorrhoeae and N. meningitidis, as well as a variety of Neisseria, Branhamella, and Moraxella species. The RapID NH system correctly identified all N. gonorrhoeae, N. meningitidis, and N. lactamica isolates, but the level of accuracy varied considerably for the remaining organisms. One strain of N. subflava was misidentified as a pathogenic Neisseria strain. The RapID NH tests were concluded in 4 h, whereas the cystine proteose peptone agar tests required up to 48 h for results to be useful. The RapID NH system is an accurate, reliable, and useful method for the identification of pathogenic Neisseria species. It has been proven that it shortens identification time and specimen turnaround time by at least 24 h.

Humans↗
PubMed1985

Rapid enzyme system for the identification of pathogenic Neisseria spp.

Gonochek II is a combination of three enzyme substrates in one tube which will give a rapid identification (30 min) of those pathogenic Neisseria spp. which can be isolated on Thayer Martin or similar selective media. Eighty isolates were tested by Gonochek II and a carbohydrate utilization method; total agreement was achieved between the two methods.

Aminopeptidases↗
PubMed1994

Utility of newer techniques for classification and identification of pathogenic anaerobic bacteria.

Results of genetic and biochemical analyses have broadened our understanding of taxonomic relationships among groups of anaerobic bacteria and have led to a better understanding of the pathogenesis of infection. Conventional bacteriologic methods are still of prime importance for the detection and identification of anaerobic pathogens. The use of nucleic acid probes has so far been restricted to research laboratories. A polymerase chain reaction-generated probe would be most useful for the rapid detection of toxigenic Clostridium difficile in feces. Probes are needed for detection of periodontopathogenic bacteria in dental plaque. Use of nucleic acid probes may become a useful adjunct to classic methodology in reference and teaching laboratories.

Bacteria, Anaerobic↗
PubMed2004

[Molecular taxonomy and identification of pathogenic fungi based on DNA sequence analysis].

Although approximately 80,000 fungi are known, less than 1% are associated with human infection. However, their taxonomy has long been insufficient. During the last decade, DNA sequence analysis was introduced to the taxonomy of pathogenic fungi. Taxonomic advances in the field of medical mycology are helping to identify the causative agents of infectious diseases accurately, facilitating diagnosis and treatment. For example, Malassezia furfur was long considered the major microflora in atopic dermatitis, yet recent studies have indicated that this is not the case, as M. furfur is taxonomically heterogeneous and consists of five species. DNA sequence analysis resolved its taxonomic heterogeneity. Similar examples can be seen in "Candida albicans and C. dubliniensis" and "Trichosporon cutaneum and T. asahii". DNA sequence analysis also enables accurate identification of fungi. At present, almost all pathogenic fungi can be identified by determining the D1/D2 26S rDNA and ITS region of rRNA gene. This paper describes the practical taxonomy and identification of pathogenic fungi based on DNA sequence analysis.

Fungi↗
PubMed1994

Haemophilus parainfluenzae endocarditis: application of a molecular approach for identification of pathogenic bacterial species.

Haemophilus parainfluenzae is both a human oropharyngeal commensal bacterium and a cause of serious invasive disease. The fastidious growth characteristics of this organism and the poor specificity of traditional methods for species identification are likely to have led to inaccuracies in the diagnosis of infections caused by H. parainfluenzae and related organisms. We report a case of H. parainfluenzae endocarditis in which confusion related to microbial identification was resolved by the analysis of 16S ribosomal RNA sequences. Rapid identification was facilitated by amplification of 16S ribosomal DNA directly from cultured cells with use of the polymerase chain reaction and by direct DNA sequence determination of the amplified product. This procedure is potentially useful for the identification of fastidious bacterial pathogens by reference laboratories.

Adult↗