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Biomedical subjects

B Dwyer

Publications and source records attributed to B Dwyer.

At least 19 recordsLinked to original sources

Emerging epidemic of community-acquired methicillin-resistant Staphylococcus aureus infection in the Northern Territory.

OBJECTIVE: To investigate the epidemiology of WA MRSA (the recently recognised Western Australian strains of methicillin-resistant Staphylococcus aureus) in the north of the Northern Territory (NT). DESIGN: Retrospective survey of data from hospital records. SETTING: Royal Darwin Hospital (a tertiary referral hospital that serves the north of the NT) between January 1991 and July 1995. SUBJECTS: All inpatients with clinical MRSA infection. OUTCOME MEASURES: Incidence of MRSA infection, classification of MRSA as WA or EA (Eastern Australian) based on antibiotic susceptibility, patient demographic details (age, sex, ethnicity, region of residence), source of infection (nosocomial or community-acquired). RESULTS: There were 125 WA MRSA and 93 EA MRSA infections, comprising 7% of all S. aureus infections. The incidence of WA MRSA infections consistently increased, while that of EA MRSA initially fell and then increased. All EA MRSA infections were nosocomial, while 50% of WA MRSA infections were community-acquired. Rates of WA MRSA infections were highest in patients from the west region of the NT, adjacent to the Kimberley region of Western Australia (WA). Community-acquired WA MRSA infections were more likely to affect Aboriginals than non-Aboriginals (relative risk [RR], 25.86; 95% confidence interval [CI], 12.51-53.47, based on population data; RR, 15.43; 95% CI, 7.85-30.32, based on admission data), as were nosocomial EA MRSA infections (RR, 2.54; 95% CI, 1.44-4.47, based on population data; RR, 2.30; 95% CI, 1.52-3.46, based on admission data). CONCLUSIONS: Changes in the epidemiology of MRSA infection in the north of the NT are consistent with the hypothesis that community-acquired WA MRSA spread into and across the NT from the Kimberley region of WA. Alternatively, crowded living conditions, hygiene difficulties and increasing use of broad spectrum antibiotics may have led to independent emergence of WA MRSA in both regions. Current infection control policies and their use in rural Aboriginal communities must be reassessed.

Adolescent

Chromoblastomycosis: a retrospective study of six cases at the Royal Darwin Hospital from 1989 to 1994.

Chromoblastomycosis, a localized chronic cutaneous and subcutaneous infection of the skin caused by pigmented fungi, is most common in the world's tropical and subtropical zones. The condition rarely occurs in Australia. We present 6 cases of chromoblastomycosis seen at the Royal Darwin Hospital, Northern Territory, from 1989 to 1994 and affecting predominantly male Caucasians ranging from 38 to 71 yrs of age. Clinically the lesions were verrucous or nodular. They mimicked basal or squamous cell carcinoma, nevi or solar keratoses. Histopathologic findings were nonspecific. The only pathognomonic finding was the presence of brown spores or sclerotic bodies within granulomata or within microabscesses in the skin.

Adult

Genetic variation in Australian spotted fever group rickettsiae.

Rickettsiae were isolated by cell culture of buffy coat blood from six patients with spotted fever from southeastern Australia and Flinders Island in Bass Strait. The isolates were genetically compared with two previous Rickettsia australis patient isolates. The genus-specific 17-kDA genes from the isolates were compared after DNA amplification and restriction fragment analysis of the amplified DNA. This comparison revealed that mainland rickettsial isolates from southeastern Australia were identical to two previous isolates of R. australis from northeastern Australia. Rickettsial isolates from Flinders Island were distinct from the mainland isolates. The 16S rRNA gene sequences from the isolates were determined and compared. The Flinders Island rickettsial agent was most closely related (0.3% structural divergence) to Rickettsia rickettsii, Rickettsia conorii, and Rickettsia slovaca. The Flinders Island rickettsial agent was 1.3 and 2.1% structurally divergent from R. australis and Rickettsia akari, respectively. The 16S rRNA gene sequence from the Flinders Island agent shows that this rickettsia is more closely related to the rickettsial spotted fever group than is R. australis. We conclude that there are two populations of spotted fever group rickettsiae in Australia and propose that the genetically distinct causative organism of Flinders Island spotted fever be designated Rickettsia honei. The extent of distribution and animal host reservoirs remain to be elucidated.

Adult

Cytotoxin production by Helicobacter pylori from patients with upper gastrointestinal tract diseases.

A cytotoxin produced by some Helicobacter pylori strains has recently been identified. The cytotoxin induces intracellular vacuolization of cultured cells. The aim of the present study was to examine the frequency of occurrence of cytotoxin-producing strains of H. pylori from subjects with upper gastrointestinal disease including nonulcer dyspepsia, gastric and duodenal ulcer disease, gastroesophageal reflux disease, and gastric cancer. Broth culture filtrates of clinical isolates of H. pylori recovered from 175 patients were used to inoculate Vero and HeLa cell monolayers for the detection of vacuolating cytotoxin activity. The results obtained demonstrated that the highest percentage of strains producing cytotoxin were found in subjects with peptic ulcer disease (gastric ulcer, 65%; duodenal ulcer, 66%; P < 0.01 compared with nonulcer dyspepsia, 38%). Of the 11 patients with gastroesophageal reflux disease, 4 of 5 patients in this group who had esophageal ulcers, were found to be infected with strains that produced cytotoxin. Three of the four patients with carcinoma of the stomach were also found to be infected with cytotoxic strains of H. pylori. With increasing severity of mucosal damage in subjects with a normal upper gastrointestinal tract, macroscopic gastritis, duodenitis, and peptic ulceration, there were corresponding increase in the proportion of strains producing cytotoxin; these increases were 32, 46, 50, and 66%, respectively. H. pylori strains from subjects with ulcer disease commonly produced vacuolating cytotoxin, suggesting that it may be a virulence factor in the pathogenesis of peptic ulcer disease.

Animals

Comparison of Pneumocystis carinii detection by toluidine blue O staining, direct immunofluorescence and DNA amplification in sputum specimens from HIV positive patients.

Pneumocystis carinii pneumonia (PCP) is the commonest opportunistic infection in AIDS patients. By using the polymerase chain reaction (PCR), specific DNA sequences can be amplified and used in diagnosis of infections such as PCP where the causative pathogen is both difficult to grow and present in low numbers. Twenty HIV positive patients were investigated for PCP. Twenty sputa (15 induced and 5 expectorated) had toluidine blue O staining, direct immunofluorescence and PCR performed for Pneumocystis carinii in a blinded fashion. PCR was performed using primers pAZ102-E 5' GATGGCTGTTTCCAAGCCCA 3' and pAZ102-H 5' GTGTACGTTGCAAAGTACTC 3' from the gene coding for Pneumocystis carinii mitochondrial ribosomal RNA with a specific 346 base-pair sequence being amplified from positive specimens. Ten of the patients had Pneumocystis carinii shown by conventional tests and PCR. Another 3 patients were positive only by PCR, all had evidence of infection with Pneumocystis carinii; the first was positive by subsequent conventional stains, the second was treated for bacterial bronchitis but had a non-resolving chest infection with PCP found on postmortem after 4 mths, the third had a typical interstitial infiltrate on CXR and responded to empiric PCP treatment. PCR is more sensitive than toluidine blue O staining and direct immunofluorescence in detecting Pneumocystis carinii in sputum from HIV patients and may become the diagnostic method of choice for PCP.

AIDS-Related Opportunistic Infections

A pseudoepidemic of Mycobacterium chelonae: contamination of a bronchoscope and autocleaner.

BACKGROUND: At fibreoptic bronchoscopy the potential exists for contamination of bronchoscopes and microbiological specimens. Patients may also be cross infected with acid fast bacilli (AFB). During a five month period, 12 bronchial wash specimens of 65 patients undergoing bronchoscopy, one bronchoscope and an autocleaner, were contaminated with the AFB, Mycobacterium chelonae (MCH). AIM: To eradicate AFB contamination of bronchoscopy specimens by identifying sources of contamination and modifying disinfection procedures. METHODS: To identify the source of contamination, samples for AFB culture were taken from three bronchoscopes, the autocleaner and water taps. To eradicate MCH contamination, the bronchoscopes were soaked in 2% glutaraldehyde overnight and flushed with 70% alcohol. Disinfection procedures were altered by using sterile water and containers in cleaning. Autocleaner use was discontinued. RESULTS: The autocleaner, one bronchoscope and 12 bronchial wash specimens were contaminated with MCH. All contaminants had similar electrophoretic banding on probing of their DNA fragments, suggesting a common clone of origin. After the alterations in disinfection procedures and despite prolonged soaking in 2% glutaraldehyde, three further contaminated wash specimens were isolated from one bronchoscope. Only after ethylene oxide sterilisation of this bronchoscope was the contamination overcome. Since then no further MCH contamination has occurred. No patient required treatment and there has been no clinical evidence of mycobacterial disease. CONCLUSION: To avoid contamination of bronchoscopy specimens with MCH, use of autoclearners should be discouraged and sterile water and containers used in cleaning procedures. If MCH contamination occurs in this setting, the bronchoscope and dismantled valve mechanism should undergo ethylene oxide sterilisation.

Bronchoalveolar Lavage Fluid

The epidemiology of tuberculosis in Victoria.

AIMS: To describe the epidemiology of Mycobacterium tuberculosis (MTb) in Victoria, Australia. METHODS: We analysed data from the Victorian Mycobacterium Reference Laboratory database and MTb notifications to Health and Community Services Victoria, for the period 1948 to 1992. RESULTS: The annual number of notified tuberculosis cases peaked at 1143 in 1954 and declined to 266 in 1992. The incidence rate decreased from 47 to 6.2 per 100,000 in the same period. The pattern of tuberculosis has changed significantly over the last two decades, reflecting the high prevalence of MTb in recent immigrants, particularly in those from South-East Asia. Among people born overseas the number of cases increased from 40% of notifications in 1970 to 80% in 1990; they are younger, more likely to be female and to have extrapulmonary disease, than their Australian-born counterparts. Between 1987 and 1991, the mean annual incidence of tuberculosis in people born in Australia and South-East Asia respectively was 1.5 and 47.5 per 100,000. People born in Vietnam, Cambodia and the Philippines had rates between 100 and 400 per 100,000. These rates have remained stable during the last six years. Almost 50% of migrant patients presented within five years, and 30% within two years, of arrival in Australia. The estimated mean annual incidence of MTb in homeless men between 1984 and June 1992 was 80 per 100,000. Resistance to all first-line drugs increased from 10.8% of all isolates in 1981 to 14.7% in 1990. The incidence of multi-drug resistant MTb (MDR-MTb) has been consistently less than 2% of isolates per year in the past 15 years. There were 14 cases of MTb and HIV between 1985 and June 1992. The mean annual incidence of MTb in people with AIDS is 1579 per 100,000 (range, 570-2420 per 100,000), with a relative risk of 236 (95% confidence interval [CI], 134-414). MDR-MTb has not been documented in HIV-infected individuals in Victoria. CONCLUSION: Tuberculosis remains an important public health concern. Groups at high risk include people born overseas, the homeless, and people infected with HIV. Physicians caring for AIDS patients should consider the diagnosis of MTb, since its incidence in AIDS patients is so high. Our data suggest possible shortcomings in current methods of screening and prophylactic treatment of migrants and refugees. The rising rate of drug resistance cannot be ignored. We should consider strategies to ensure judicious use of anti-tuberculous drugs by physicians, and optimal patient compliance. These issues are critical to the future of tuberculosis control in Victoria.

Adolescent

Spotted fever group rickettsial infection in south-eastern Australia: isolation of rickettsiae.

Flinders Island spotted fever (FISF), a spotted fever group (SFG) rickettsial disease first described in 1991, occurs in south-eastern Australia. The isolation of the aetiological agent is described for the first time having been obtained from the blood of two patients. An additional 22 cases are also reported. Of these patients four had positive initial serology, and 20 showed seroconversion (using Rickettsia australis as antigen). Acute phase blood specimens taken from seven patients caused neonatal mice to seroconvert to R. australis and a blood specimen from one of these patients (and one other) yielded rickettsiae. A field survey for possible reservoir and vector animals on Flinders Island, Tasmania and in Gippsland, Victoria (both in south-eastern Australia) yielded 217 vertebrates and 1445 invertebrate ectoparasites, mostly ticks. Ixodes cornuatus from humans and dogs in Gippsland produced seroconversion to SFG rickettsia when inoculated into mice but no invertebrate pools from Flinders Island produced seroconversion in mice. Haemolymph from an individual I. cornuatus removed from a human in Gippsland, yielded a SFG rickettsia on tissue culture. Sera from several species of native vertebrates, especially the bush rat, Rattus fuscipes, were positive for antibodies to SFG rickettsia.

Adolescent

DNA restriction fragment analysis to define an extended cluster of tuberculosis in homeless men and their associates.

A cluster of tuberculosis in men associated with shelters for the homeless in Melbourne, diagnosed between April 1984 and July 1991, was reviewed with respect to the genetic relatedness of the infecting strains. Relatedness was determined by examination of Southern blot analyses of restriction enzyme-digested genomic DNA, using probes for repetitive sequences. From the initial 24 cases selected, isolates were available from 19, and 18 were identical. A total of 571 other Australian and Solomon Islands strains were examined for relatedness to these strains. Nine additional case strains were found to be identical; all were from recent Melbourne residents and at least 7 were epidemiologically related to the original case cluster. The identification of a single infecting strain of Mycobacterium tuberculosis in these 27 cases suggests that reactivation disease in this group may not be as important as infection or reinfection from another case or cases.

Adult

Isolation of a DNA probe for identification of Mycobacterium kansasii, including the genetic subgroup.

In order to develop a DNA-based assay to identify all Mycobacterium kansasii clinical isolates, a specific DNA probe was isolated in plasmid p6123. A total of 145 M. kansasii clinical isolates were collected from several countries and were examined with three probes by DNA hybridization. Of the 145 isolates, 115 (79%) were positive with the previously described probe pMK1-9 (Z. H. Huang, B. C. Ross, and B. Dwyer, J. Clin. Microbiol. 29:2125-2129, 1991), 129 (88%) were positive with the commercial Accu-probe assay (Gen-Probe), and 145 (100%) were positive with the p6123 probe. Southern blot analysis of EcoRI-digested M. kansasii chromosomal DNA with p6123 revealed that all Accu-probe-positive M. kansasii strains exhibited a 3-kb fragment, whereas all Accu-probe-negative M. kansasii strains displayed DNA fragments of variable molecular sizes. These results indicate that, unlike the previously described probes for M. kansasii, the fragment cloned into p6123 identified all 145 biochemically typical strains tested and provides an ideal target for future DNA-based speciation assays.

Base Sequence

A new trichrome-blue stain for detection of microsporidial species in urine, stool, and nasopharyngeal specimens.

Detection of microsporidia in clinical specimens has relied on electron microscopy, histology, or staining. This article describes further alterations to the modified trichrome staining method which make it easier to identify microsporidial spores. The changes are a decrease in the phosphotungstic acid level and the substitution of a colorfast counterstain, aniline blue, for the fast green of the original stain. The modified stain provides good contrast between microsporidial spores and background material including human and fungal cells. Stool specimens from 139 human immunodeficiency virus-seropositive patients revealed that 5 patients were infected with Enterocytozoon bieneusi and 6 patients had larger spores. Thin-section electron microscopy of the larger spores showed a structure consistent with that of either Encephalitozoon or Septata species. Three of the patients with Encephalitozoon- or Septata-like species had disseminated infection, with spores detected in nasopharyngeal aspirates and urine samples.

AIDS-Related Opportunistic Infections

Rapid, simple method for typing isolates of Mycobacterium tuberculosis by using the polymerase chain reaction.

To develop a molecular typing method for Mycobacterium tuberculosis based on the polymerase chain reaction, oligonucleotide primers were designed to the ends of the insertion sequence IS6110 in an attempt to amplify DNA between clusters of this element on the genome. Although in many strains the copy number of this element is low and is distributed throughout the genome, most strains examined produced a banding pattern which varied between isolates including strains with one copy of IS6110. With strains isolated from patients in epidemiologic clusters of tuberculosis, the banding patterns were similar within each cluster but distinct from those in strains from different clusters. Similarly, multiple isolates from the same patient yielded a consistent banding pattern. Sequencing of four polymerase chain reaction products revealed that amplification was occurring between copies of IS6110 in two of the products and from a single copy of IS6110 to a nonspecific priming site in the other two. This technique provides a rapid and simple means of typing M. tuberculosis isolates for epidemiologic studies.

Bacterial Typing Techniques

Mycobacteria in stool specimens: the nonvalue of smears for predicting culture results.

A previous recommendation suggests that stool be cultured for mycobacteria only if the smear is positive. We have correlated smear and culture results of 2,176 stool specimens submitted for mycobacterial culture. The sensitivity, specificity, and positive and negative predictive values for smears to predict culture results are 34, 99, 90, and 87%, respectively. We recommend that the stool smear not be used as a screening technique to decide which specimens from at-risk patients should be cultured because it lacks the necessary sensitivity.

AIDS-Related Opportunistic Infections

Characterization of Mycobacterium tuberculosis strains from Vietnamese patients by Southern blot hybridization.

A total of 41 Mycobacterium tuberculosis strains from patients of Vietnamese origin were analyzed by Southern blot hybridization with two different probes, IS6110 (Otal, I., et al., J. Clin. Microbiol. 29:1252-1254, 1991; Ross, B. C., et al., J. Clin. Microbiol. 30:942-946, 1992; Thierry, D., et al., J. Clin. Microbiol. 28:2668-2673, 1990; van Soolingen, D., et al., J. Clin. Microbiol. 29:2578-2586, 1991) and pTBN12 (Ross, B. C., et al., J. Clin. Microbiol. 30:942-946, 1992). The restriction fragment patterns of nine of these strains were virtually identical when the pTBN12 probe was used; five strains had a single copy of IS6110, and four strains failed to hybridize with the IS6110 probe. This relatively high frequency of strains with no or one copy of IS6110 suggests that the usefulness of IS6110 for epidemiological study may be limited in certain populations.

Australia

Comparative evaluation of detection assays for Chlamydia trachomatis.

The performances of a commercial nucleic acid hybridization test (Gen-Probe Pace 2 Chlamydia trachomatis) and two commercial enzyme immunoassays (EIAs) (Abbott Chlamydiazyme and Pharmacia Chlamydia EIA) were evaluated against cell culture for the detection of C. trachomatis infection, with cervical swabs obtained from 1,037 women visiting a public sexual health center. The positivity rate by cell culture was 4.7%. Sensitivity and specificity for each test were as follows: Gen-Probe, 95.8 and 98.3%; Chlamydiazyme, 80.4 and 99.3%; Pharmacia EIA, 80.8 and 99.1%. Analysis of discrepant results with probe confirmation assay (Gen-Probe) and direct immunofluorescence (Syva Microtrak) revealed 12 cases of C. trachomatis infection for which culture was negative, resulting in the definition of a true-positive case as opposed to a culture positive. The positivity rate by true-positive definition was 5.9%, and sensitivity and specificity for each test were as follows: Gen-Probe, 96.7 and 99.6%; Chlamydiazyme, 77.5 and 100%; Pharmacia EIA, 77.0 and 100%; cell culture, 80.0 and 100%. We conclude that the Gen-Probe Pace 2 C. trachomatis test is a sensitive and specific alternative to cell culture for the detection of C. trachomatis.

Bacteriological Techniques

Identification of an insertion sequence-like element in a subspecies of Mycobacterium kansasii.

Analysis of a genomic DNA clone library of a strain from the genetic subspecies of Mycobacterium kansasii determined the existence of a repetitive insertion sequence-like element. The element is 947 bp long and is present in a minimum of 1 to 11 copies per genome. Similar to insertion sequences, it contains a 3-bp (TAG) direct repeat at its extremities and a transcription promoter-like sequence. In addition, for one of the clones sequenced, a potential cointegrate formation, a characteristic frequently observed with insertion sequences, was revealed. This insertion sequence does not contain short inverted repeats near the ends or a large open reading frame to code for a transposase enzyme. Its host range is restricted to a previously described genetic subspecies of M. kansasii and is not present in typical M. kansasii or other mycobacterial species. When used as a probe for Southern blot hybridization, significant heterogeneity between different isolates of the M. kansasii subspecies was observed. This repeated element will be useful in further studies on the characterization, diagnosis, and epidemiology of M. kansasii.

Base Sequence