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N F Lightfoot

Publications and source records attributed to N F Lightfoot.

At least 37 records · Page 2Linked to original sources

Rapid identification of species within the Mycobacterium tuberculosis complex by artificial neural network analysis of pyrolysis mass spectra.

An artificial neural network (ANN) was trained to distinguish between Mycobacterium tuberculosis and M. bovis with averaged pyrolysis mass spectra from duplicate subcultures of four strains of each of these species, each pyrolysed in triplicate. Once trained, the ANN was interrogated with spectrum data from the original organisms (the "training set") and from 26 other mycobacterial isolates (the "challenge set") of the M. tuberculosis complex (MTBC). Eight strains of M. bovis and 13 of M. tuberculosis, whether sensitive or variously resistant to antituberculosis drugs, were identified in agreement with conventional identification. Four strains of "M. africanum" were identified as M. bovis. Of two atypical M. tuberculosis strains from South India, one was identified as M. tuberculosis and the other as M. bovis. Six strains of BCG proved heterogeneous; two gave equivocal identifications, three were identified as M. bovis and one was identified as M. tuberculosis.

Humans↗

Isolation and purification of Aeromonas sobria cytotonic enterotoxin and beta-haemolysin.

Aeromonas sp., grown in tryptone soya broth supplemented with yeast extract, 0.6%, pH 7.5, and incubated with agitation at 100 oscillations/min for 15 h at 37 degrees C produced optimal amounts of beta-haemolysin and cytotonic enterotoxin. More prolonged incubation resulted in the loss of enterotoxic activity and anion exchange chromatographic analysis indicated the presence of a moiety capable of breaking down the toxin. Anion exchange fast protein liquid chromatography resulted in a single peak of haemolytic activity and two peaks with enterotoxic activity. The cytotonic enterotoxin was purified from the fraction most active in the infant mouse assay; the second peak, which did not cross-react immunologically, may represent a second cytotonic enterotoxin. Neither peak was observed in the chromatographic fractions of filtrates from strains devoid of activity in the infant mouse assay. Purified enterotoxin, estimated to have a mol. wt of 15 kDa by SDS-PAGE, caused fluid accumulation in the infant mouse assay, was non-haemolytic to rabbit erythrocytes, caused an increase in cAMP activity in tissue culture cells and did not cross-react immunologically with components of cholera toxin or the whole toxin. Purified beta-haemolysin had an estimated mol. wt of 55 kDa, lysed rabbit erythrocytes and did not cause fluid accumulation in the infant mouse test.

Aeromonas↗

External quality assessment in water microbiology: statistical analysis of performance.

A UK-based scheme of water microbiology assessment requires participants to record counts of relevant organisms. Not every sample will contain the target number of organisms because of natural variation and therefore a range of results is acceptable. Results which are tail-end (i.e. at the extreme low or high end of this range) could occasionally be reported by any individual laboratory by chance. Several tail-end results might imply a laboratory problem. Statistical assessment is done in two stages. A non-parametric test of the distribution of tail-end counts amongst laboratories is performed (Cochran's Q) and, if they are not random, then observed and expected frequencies of tail-end counts are compared to identify participants who may have reported excessive numbers of low or high results. Analyses so far have shown that laboratories find high counts no more frequently than would be expected by chance, but that significant clusters of low counts can be detected among participants. These findings have been observed both in short-term and in long-term assessments, thus allowing detection of new episodes of poor performance and intermittent problems. The analysis relies on an objective definition of tail-end results. Working definitions are presented which should identify poor performance in terms of microbiological significance, and which allow fair comparison between membrane-filtration and multiple-tube techniques. Smaller differences between laboratories, which may be statistically significant, will not be detected. Different definitions of poor performance could be incorporated into future assessments.

Colony Count, Microbial↗

Application of pyrolysis mass spectrometry to the investigation of outbreaks of food poisoning and non-gastrointestinal infection associated with Bacillus species and Clostridium perfringens.

Eighteen isolates of Bacillus species and 15 of Clostridium perfringens, all of which had been associated with outbreaks of either food poisoning or non-gastrointestinal infection (NGI), were examined for relatedness by pyrolysis mass spectrometry (PyMS). The PyMS-analysis correctly clustered all the groups of epidemiologically related isolates of both genera, and distinguished all the single, epidemiologically unrelated isolates of the same species. PyMS is a simple, rapid and inexpensive technique which can provide useful and accurate inter-strain comparisons within both the Bacillus and Clostridium genera in complete accord with conventional serological typing results.

Bacillaceae Infections↗

Nosocomial infection with Clostridium difficile investigated by pyrolysis mass spectrometry.

Fifty-eight isolates of Clostridium difficile from two distinct outbreaks were examined for inter-strain similarity by pyrolysis mass spectrometry (PMS). The first outbreak began on a geriatric acute unit and spread to a long stay geriatric facility. PMS analysis showed that most isolates from both sites were indistinguishable. Isolates obtained in the preceding year from the long stay facility were found to be closely similar to these outbreak isolates. In the second, smaller outbreak on a female medical ward in another general hospital, PMS again showed that a single strain was probably responsible. Representative isolates from these two different outbreaks were shown to be distinct. The ability to compare rapidly large numbers of isolates of C. difficile makes PMS attractive for initial screening in suspected outbreaks, providing important information for outbreak management and allowing conventional typing methods to be concentrated on relevant isolates.

Aged↗

Rapid differentiation of Mycobacterium xenopi from mycobacteria of the Mycobacterium avium-intracellulare complex by pyrolysis mass spectrometry.

Thirty four cultures of slow growing, Tween-80 negative mycobacteria were analysed by pyrolysis mass spectrometry. The results showed that pyrolysis mass spectrometry could positively distinguish strains of Mycobacterium xenopi from those of the Mycobacterium avium-intracellulare (MAI) complex. Pyrolysis mass spectrometry may be a useful technique for the rapid characterisation of non-tuberculous mycobacteria in such clinical settings as their isolation from immunocompromised patients-for example, those with AIDS.

Hot Temperature↗

Legionella pneumophila species identification using a commercial latex agglutination kit: a potential cross-reaction problem with serogroup 12.

Seventy-nine strains of legionella spp. and 13 non-legionella isolates were tested using a commercial latex agglutination kit. Cross-reactions were observed for all strains of Legionella pneumophila serogroup 12 examined between latex particles coated with rabbit antibodies raised against L. pneumophila serogroup 1 and those coated with rabbit antibodies to serogroups 2-14. Strains of L. pneumophila serogroup 1 did not cross-react with latex particles directed against L. pneumophila serogroups 2-14. This suggests that L. pneumophila serogroup 12 shares common surface antigens with L. pneumophila serogroup 1, and that conversely serogroup 1 isolates do not share major surface antigen determinants with serogroups 2-14. All other isolates tested gave expected results.

Antigens, Bacterial↗

Differentiation between mycobacteria of the Mycobacterium tuberculosis complex by pyrolysis mass spectrometry.

32 isolates belonging to the Mycobacterium tuberculosis complex were examined by pyrolysis mass spectrometry (PyMS). This technique demonstrated that recent clinical isolates of M. africanum were indistinguishable from those of M. bovis and together formed a homogeneous group distinct from M. tuberculosis. Isolates of BCG were heterogeneous and more closely related to laboratory-adapted strains of M. tuberculosis than to recent isolates of either M. tuberculosis or M. bovis. PyMS is a simple and inexpensive technique which gives interesting information on the relationships between members of the M. tuberculosis complex and can make the clinically important distinction between strains of M. bovis and M. tuberculosis accurately and much more rapidly than conventional techniques.

Mass Spectrometry↗

Strain differentiation of nosocomial isolates of Pseudomonas aeruginosa by pyrolysis mass spectrometry.

Pyrolysis mass spectrometry (PyMS) was used for rapid interstrain comparison of Pseudomonas aeruginosa isolates from a small outbreak of sternal wound infections in a cardiothoracic surgical ward. The PyMS results were compared with those obtained by conventional O-serotyping. All the isolates were phage non-typable. Evidence obtained from PyMS of identity/non-identity between isolates correlated completely with that obtained by O-typing and correctly identified those isolates comprising the epidemic strain, in one instance in advance of O-serotyping. The speed and versatility of PyMS make it an attractive technique for the initial screening of isolates from nosocomial outbreaks of infection with P. aeruginosa and other organisms.

Bacterial Typing Techniques↗

Inter-strain comparison by pyrolysis mass spectrometry in the investigation of Staphylococcus aureus nosocomial infection.

Pyrolysis mass spectrometry (PyMS) was used to examine isolates of Staphylococcus aureus from an outbreak of wound infections on a cardiothoracic surgical unit, some of which were thought to have been related to a point-source in the operating theatre. The PyMS results were compared with the results of phage typing. Both methods suggested that a single strain of S. aureus, of phage pattern 29/52/52A/79/80/81, was responsible for some of the wound infections, but PyMS also identified two patients with phage non-typable isolates. Phage typing indicated four staff members as possible carriers of the epidemic strain, but PyMS indicated only two. Epidemiological enquiry confirmed that one of the two members of staff identified by both methods was likely to have been the source of the theatre-based infection. PyMS is a rapid and relatively inexpensive technique for the investigation of nosocomial S. aureus infection and was more discriminatory than phage typing in this instance.

Carrier State↗

Use of ciprofloxacin to control a Salmonella outbreak in a long-stay psychiatric hospital.

An outbreak of diarrhoea due to Salmonella virchow phage type 8 occurred in a major block of a large psychiatric hospital. The two other major blocks of the hospital remained unaffected. No source of infection was identified and the epidemiological investigations pointed to cross-infection as the mode of transmission. Infection control measures were implemented at an early stage but a total of 55 patients and four members of staff were affected. The disease was self-limiting and of short duration. During the course of the outbreak, Salmonella typhimurium phage type 66 also emerged as a pathogen and was isolated from the stools of symptomatic cases with diarrhoea. Owing to the size of the hospital, the lack of facilities for infection control, the nature of the patients' illnesses and an increasing number of affected patients, it was decided to treat all symptomatic and asymptomatic patients with oral ciprofloxacin 500 mg twice daily for 7 days. A quantitative stool culture for salmonellae was undertaken on days 3, 7, 10, 30, 60 and 90 from the start of treatment. All patients' and staff's stool cultures became negative on day 7 and remained negative for a period of 6 months during follow-up. We conclude that cross-infection outbreaks due to Salmonella spp. in psychiatric and geriatric hospitals can be controlled by isolation in a designated ward and supportive therapy, together with ciprofloxacin treatment.

Adult↗

Rapid inter-strain comparison by pyrolysis mass spectrometry in nosocomial infection with Xanthomonas maltophilia.

Seventeen strains of Xanthomonas maltophilia and one strain of Pseudomonas cepacia were examined by pyrolysis mass spectrometry (PYMS). The Xanthomonas strains comprised 11 clinical and environmental isolates from a suspected outbreak of colonization and infection on a heart-lung transplant intensive care unit, two strains from patients elsewhere in the same hospital and four strains from a national reference collection. The single isolate of Pseudomonas cepacia was from a sink in the same affected intensive care unit. A series of discriminant analyses performed on the PYMS-derived data showed that, whereas six strains of Xanthomonas from the respiratory tract, blood and ventilatory equipment of one of the affected patients were indistinguishable, all the other isolates were distinct. The results of PYMS rapid inter-strain comparison were in accord with those of an epidemiological investigation which suggested that the episode was due to unauthorized reuse of disposable nebulizers and not to cross-infection between patients. Pyrolysis mass spectrometry with rapid data analysis is a potentially useful technique for the investigation of nosocomial infections due to organisms such as X. maltophilia.

Bacteriological Techniques↗

Rapid inter-strain comparison by pyrolysis mass spectrometry of coagulase-negative staphylococci from persistent CAPD peritonitis.

Pyrolysis mass spectrometry (PyMS) was used as a method of rapid inter-strain comparison of 19 isolates of coagulase-negative staphylococci from episodes of CAPD peritonitis. Thirteen isolates were from multiple, but distinct, episodes of peritonitis in 6 patients and the remaining 6 isolates were from 6 patients with single episodes. The results, expressed in terms of identity/non-identity of strains, were compared with those obtained using an established typing system comprising an extended antibiogram, determination of biotype and plasmid profile analysis. The PyMS results for inter-strain comparison were in agreement with the reference typing scheme results. PyMS can be used in this setting to rapidly obtain evidence that persistent infection is/is not likely to be due to the same organism, although it cannot be used for formal typing. The results by both methods showed that serial, apparently distinct, episodes of peritonitis over periods as long as 120 days may be due to the same strain of coagulase-negative staphylococcus. Clinically based distinctions between recurrence of infection (same strain) and re-infection (different strains) may not always be supported by the microbiological evidence.

Coagulase↗

Incrimination of an environmental source of a case of Legionnaires' disease by pyrolysis mass spectrometry.

Following a case of Legionnaires' disease Legionella pneumophila of the same serogroup was isolated from the hot water system of the hotel which had been epidemiologically implicated as the source of the infection. Pyrolysis mass spectrometry (PyMS) was used to compare 11 isolates from the hotel water sample with the patient's strain. Epidemiologically unrelated isolates, both clinical and environmental, of the same serogroup and monoclonal antibody type of L. pneumophila were included in the same analysis, together with relevant reference strains. The patient strain was shown to be indistinguishable from seven of the hotel water isolates, but clearly different from other unrelated clinical isolates, the reference strains and some of the other environmental isolates. PyMS is a rapid and cheap method for inter-strain comparison for L. pneumophila.

Aged↗