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Biomedical subjects

M Glennon

Publications and source records attributed to M Glennon.

32 records · Page 2Linked to original sources
PubMed1995

Evaluation of a PCR assay for detection of Mycobacterium tuberculosis in clinical specimens.

A polymerase chain reaction (PCR) assay for detection of M. tuberculosis was optimized for application to clinical specimens, which were prepared for amplification by boiling in buffer. The buffer contained a synthetic DNA fragment to determine if DNA amplification from the individual prepared specimens was subject to inhibition because of substances present in the specimen, or by the process of specimen preparation or storage. The PCR test was less sensitive than direct microscopy (75% as against 87.5%) and had a specificity of 97%. Invalid results due to inhibition of amplification occurred in 12% of specimens. Incorporation of the internal standard into the specimen preparation buffer ensures that any step in the process which inhibits DNA amplification is detected in the failure of amplification of the internal standard. The use of internal standard in this way should be considered in developing diagnostic protocols.

Base Sequence↗
PubMed1995

The sensitive detection of fluorescently labelled PCR products using an automated detection system.

The polymerase chain reaction plays a central role in many detection assays and methods to improve the sensitivity and specificity of these detection systems are constantly being explored. In this study we investigated the use of an automated laser fluorescent system (ALF) in the context of DNA-based diagnostics for pathogenic bacteria. PCR products were generated using species-specific primer sets, one of which was labelled with a 5' fluorescein. PCR products with a fluorescent label were detected on line with an ALF DNA sequencer and the sensitivity of detection was found to be comparable to that for DNA probe hybridization with a radioactive probe. The technology was successfully applied to the detection of Mycobacterium tuberculosis supplemented into sputum samples and to the detection of listeria in paraffin-embedded tissue samples.

Base Sequence↗
PubMed1995

Multiplex PCR for identifying mycobacterial isolates.

AIMS: To develop a multiplex polymerase chain reaction (PCR) method to facilitate identification of mycobacterial isolates. METHODS: Type strains of 14 species of mycobacteria and 56 clinical isolates were lysed by boiling in TE Triton. The lysate (5 microliters) was used directly in a PCR reaction incorporating three pairs of PCR primers expected to amplify fragments from the genome of (a) all mycobacteria, (b) Mycobacterium tuberculosis complex only and (c) M avium only. PCR products were visualised by electrophoresis on agarose gels. RESULTS: Multiplex PCR applied to 14 type strains yielded patterns on electrophoresis which permitted identification of the mycobacterial isolates as M tuberculosis complex, M avium or as mycobacteria other than the former. The identification of 56 clinical isolates by multiplex PCR was consistent with other methods and was accomplished in less than one working day. CONCLUSIONS: This method may facilitate rapid and convenient identification of most clinical isolates of mycobacteria by PCR and gel electrophoresis. Further evaluation is warranted.

Base Sequence↗
PubMed1994

PCR based fingerprinting of Enterobacter cloacae.

An outbreak of lower respiratory tract infection with Enterobacter cloacae occurred in an intensive care unit in a university teaching hospital. Random amplification of polymorphic DNA (RAPD) was used to assist in the investigation of the outbreak. The technique was readily applied to this organism and permitted differentiation between strains which had identical biochemical profiles and antibiograms. The versatility of this technique makes it attractive for use in hospitals where fingerprinting of any one of the many Gram-negative rods associated with nosocomial infection may be required from time to time.

Cross Infection↗
PubMed1994

The ribosomal intergenic spacer region: a target for the PCR based diagnosis of tuberculosis.

SETTING: DNA Diagnostics, The National Diagnostics Centre and University College Hospital, Galway, Republic of Ireland. OBJECTIVE: To investigate the possibility of developing a DNA probe to distinguish the members of the Mycobacterium tuberculosis complex and develop an assay for the detection of M. tuberculosis in sputum. DESIGN: The ribosomal intergenic spacer regions from the members of the M. tuberculosis complex were sequenced and analysed for the ability of this region to provide sequence diversity for DNA probe development. RESULTS: The 100% sequence homology in this region precluded the development of a probe to distinguish the individual members of the M. tuberculosis complex. However it was possible to develop a DNA probe that could specifically detect only the members of the M. tuberculosis complex. CONCLUSION: A specific DNA probe was developed for the detection of the M. tuberculosis complex and its application to M. tuberculosis detection in sputum was demonstrated.

Base Sequence↗
PubMed1991

Role of normal serum in the binding of lipopolysaccharide to IgG fractions from rabbit antisera to Escherichia coli J5 and other gram-negative bacteria.

Because lipopolysaccharide (LPS) bound to lipoprotein is less active than unbound LPS in multiple assay systems, the binding of radiolabeled LPS to lipoproteins in sera prepared from normal rabbits and rabbits made hyperimmune to Escherichia coli J5 were compared. LPS-lipoprotein binding in hyperimmune sera to E. coli J5 was not greater than that in normal serum as assessed by ultracentrifugation, but more LPS was precipitated from hyperimmune antisera than normal sera under conditions designed to precipitate LPS-lipoprotein complexes with calcium and dextran. Radiolabeled LPS was precipitated by delipidated antisera and fractions of IgG purified by anion exchange chromatography, but the precipitation was dependent on the presence of normal serum in the reaction mixture. These data suggest that a fluid-phase RIA done in the presence of normal serum may facilitate the detection of IgG in antisera raised to E. coli J5 that binds to heterologous smooth LPS.

Animals↗
PubMed1991

The 16s/23s ribosomal spacer region as a target for DNA probes to identify eubacteria.

Variable regions of the 16s ribosomal RNA have been frequently used as the target for DNA probes to identify microorganisms. In some situations, however, there is very little sequence variation observed between the 16s rRNA genes of closely related microorganisms. This study presents a general method to obtain species-specific probes using the spacer (intergenic) region between the 16s and 23s rRNA genes. The overall strategy is analogous to that which has previously been developed for the variable regions of the 16s rRNA genes. Sequence analysis of the 16s rRNA and 23s rRNA coding sequences flanking the spacer regions resulted in the design of PCR primers that can be used to amplify the spacer regions of a wide range of eubacteria. Sequencing the amplified spacer region then gives rise to the information that can be used to select specific DNA sequences for use as a DNA probe or for the generation of specific PCR primers to a microorganism of interest. In this study the approach to develop specific DNA markers for members of the genus Clostridium is described in detail. A specific DNA oligonucleotide probe and PCR primers have been designed for Clostridium perfringens that distinguish it from other organisms in the genus.

Bacteria↗
PubMed1990

Garfish injury.

Explore the source record for details and available documents.

Adult↗
PubMed1990

Yaws in the Solomon Islands.

Yaws is a chronic, relapsing, non-venereally transmitted disease caused by Treponema pertenue. As a result of the WHO mass treatment campaign of the late 1950s, the prevalence in the Solomon Islands fell dramatically. Here the disease was thought to have been eradicated until an outbreak occurred in 1981. In 1984 a mass treatment survey following modified WHO guidelines was carried out. Subsequent to this campaign, yaws recurred and in 1987 a further treatment survey was required. Two observations were made as a result of our recent experience in controlling yaws in the Solomon Islands. (1) The disease appears to be attenuated. (2) WHO control policy may now be an inappropriate method for dealing with yaws in the Solomon Islands and should be replaced by a method which is integrated into the existing primary health care (PHC) structure.

Adolescent↗
PubMedDate not supplied

Eating difficulties in patients with probable dementia of the Alzheimer type.

Seventy-three institutionalized patients suffering from probable dementia of the Alzheimer type (DAT) were surveyed for the presence of eating difficulties. Among 71 patients fed by natural means four different groups emerged: (1) patients who fed themselves (n = 17, 23.9%), (2) patients who had to be fed but posed no other eating problem (n = 13, 18.3%), (3) patients who refused food although they were able to swallow it (n = 18, 25.4%), and (4) patients who choked on liquid and/or solid food, some of whom also refused food (n = 23, 32.4%). Patients who fed themselves were in a less advanced stage of the disease than those who did not, and their average body weight was equal to the ideal weight. The remaining three groups, ie, those with different eating problems, did not differ in mean severity of DAT, and their body weights were significantly lower. The mortality rate during 2 years following the survey was similar in all four groups of patients, although tube feeding was used in only one case. The mortality rate was also similar in patients whose body weights were 20% or more below the median weight for their age, and in patients whose relative body weight was higher. The results of this study suggest that eating difficulties occur in a majority of institutionalized DAT patients, but can be managed without resorting to tube feeding.

Activities of Daily Living↗