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Biomedical subjects

J E Clarridge

Publications and source records attributed to J E Clarridge.

At least 19 recordsLinked to original sources
PubMed1995

An outbreak of Burkholderia (formerly Pseudomonas) cepacia respiratory tract colonization and infection associated with nebulized albuterol therapy.

OBJECTIVE: To investigate an outbreak of Burkholderia (formerly Pseudomonas) cepacia respiratory tract colonization and infection in mechanically ventilated patients. DESIGN: A retrospective case-control and bacteriologic study. SETTING: Veterans Affairs medical center. PATIENTS: 42 mechanically ventilated patients who developed respiratory tract colonization or infection with B. cepacia and 135 ventilator-dependent controls who were not colonized and did not develop infections. MEASUREMENTS: Clinical and demographic data; benzalkonium chloride concentrations and pH levels in albuterol sulfate solutions; repetitive-element polymerase chain reaction (PCR)-mediated molecular fingerprinting on eight patient isolates and three environmental B. cepacia isolates that were available for study. RESULTS: 42 patients had B. cepacia respiratory tract colonization or infection. Observation of intensive care unit and respiratory care personnel showed faulty infection control procedures (for example, the same multiple-dose bottle of albuterol was used for many mechanically ventilated patients). More case patients (39 [92.9%]) than controls (95 [70.4%]; P = 0.006) received nebulized albuterol, and case patients (67.5 treatments) received more treatments than controls (18 treatments; P < 0.001). In-use albuterol solutions had pH values that were unstable, and benzalkonium chloride concentrations declined over time to levels capable of supporting bacterial growth. Medication nebulizers and in-use bottles of albuterol harbored B. cepacia. Molecular fingerprints of patient isolates and environmental B. cepacia isolates were identical using repetitive-element PCR. No further isolates of B. cepacia were identified after institution of appropriate infection control procedures. CONCLUSIONS: Multiple-dose medications and reliance on benzalkonium chloride as a medication preservative provide a mechanism for nosocomial spread of microorganisms, particularly if infection control procedures are not carefully followed. Repetitive-element PCR is a useful fingerprinting technique for molecular epidemiologic studies of B. cepacia.

Aged↗
PubMed1995

Transport and storage of Helicobacter pylori from gastric mucosal biopsies and clinical isolates.

Various transport and storage conditions for the recovery of Helicobacter pylori from gastric biopsies were evaluated. Gastric mucosal biopsies from 16 Helicobacter pylori-infected patients were stored in cysteine-Albimi medium containing 20% glycerol in a refrigerator (4 degrees C) for 1 and 2 weeks and in a -20 degrees C laboratory freezer for 4 and 12 weeks. Two clinical isolates were stored in saline, Stuart's transport media, cysteine-Albimi broth with 20% glycerol, brucella broth with 20% glycerol and skim milk with 17% glycerol at room temperature, 4 degrees C, -20 degrees C and -70 degrees C. Storage at 4 degrees C for 1 and 2 weeks resulted in Helicobacter pylori recovery from 81% and 19% of biopsies, respectively. Storage at -20 degrees C yielded Helicobacter pylori recovery in 100% and 57% after 4 and 12 weeks, respectively. At room temperature after 6 h, the Helicobacter pylori titer was reduced. The best storage media for frozen isolates were skim milk/glycerol, brucella broth/glycerol and cysteine-Albimi/glycerol (in descending order). Recovery was better at -70 degrees C than -20 degrees C.

Biopsy↗
PubMed1995

Recovery of Coccidioides immitis from blood and abscess fluid using the BacT/alert system.

We report the recovery of Coccidioides immitis from the blood and abscess fluid of two separate patients by using two automated blood culture systems. In the first case, an aspirate from a neck abscess containing C. immitis spherules was serially diluted and inoculated into liquid media used by the BacT/Alert and the Bactec NR660 blood culture systems. BacT/Alert bottles inoculated with 10(5), 10(4), 10(3), 10(2), 10 and two spherules produced a positive signal at 19, 24, 35, 42, 57, and 62 h postinoculation, respectively. Bactec NR660 bottles containing > 10(2) shperules and 10 spherules produced a positive signal after approximately 72 and 96 h of incubation, respectively. In the second case, a blood specimen incubated in BacT/Alert blood culture both was signaled positive after 82 h of incubation. No organisms were detected by Gram stain of the broth, but C. immitis grew after blind subculture. Our observations demonstrate that these rapid blood culture systems are capable of supporting growth of C. immitis. To our knowledge, this report is the first to detect C. immitis by these blood culture systems.

Adult↗
PubMed1995

Observations on the pathological spectrum and clinical course of microsporidiosis in men infected with the human immunodeficiency virus: follow-up study.

The purposes of this research were to determine whether microsporidian infection is accompanied by structural alterations of the duodenal mucosa and to characterize the clinical and morphological evolution of intestinal microsporidiosis. In a previous systematic electron microscopic evaluation of mucosal biopsy specimens from patients with human immunodeficiency virus infection, we detected Enterocytozoon bieneusi in the duodenal tissue of 31 men. This report concerns a follow-up study of these 31 men, who underwent repeated clinical and laboratory evaluations, including repeated duodenal biopsies. Eighteen men had chronic diarrhea at enrollment and 13 had no diarrhea. The CD4+ cell counts (per mm3) ranged from 10 to 660 and were normal for three men. E. bieneusi infection was accompanied by a wide spectrum of histopathology of duodenal tissue; over a mean follow-up of 15 months, no consistent change in duodenal mucosal morphology was observed, and diarrhea did not invariably develop. These observations indicate that diarrhea and severe villus injury do not invariably accompany infection. Further studies are needed to determine the relationship between E. bieneusi infection and clinical illness.

AIDS-Related Opportunistic Infections↗
PubMed1995

In vitro evaluation of the role of humoral immunity against Bartonella henselae.

The contribution of humoral immunity against Bartonella henselae was evaluated by examining the in vitro bactericidal activity of sera and the ability of these microorganisms to activate complement and stimulate phagocytosis and an oxidative burst in polymorphonuclear leukocytes. The organism was killed by complement-mediated cytolysis. Complement activation preferentially proceeded by the alternative pathway. The presence of specific antibodies did not increase the serum bactericidal activity or complement activation. However, phagocytosis and the subsequent production of oxygen radicals, evaluated by flow cytometry, were significantly enhanced in the presence of bacteria previously opsonized with immune sera.

Adult↗
PubMed1995

Genomic fingerprinting of Bartonella species by repetitive element PCR for distinguishing species and isolates.

Repetitive-element PCR (rep-PCR) with primers based on repetitive extragenic palindromic (REP) and enterobacterial repetitive intergenic consensus (ERIC) repeated DNA sequences was used for genomic finger-printing of Bartonella species. This technique was applied by using either extracted genomic DNA or preparations of whole bacterial cells directly. PCR fingerprints with either the REP-based primers (REP-PCR) or primers based on the ERIC repeat (ERIC-PCR) revealed species-specific band patterns for the various Bartonella isolates. DNA fingerprints obtained from rep-PCR of extracted genomic DNA or from preparations of whole cells yielded comparable patterns. ERIC-PCR banding patterns were less complex than those obtained by REP-PCR but allowed better discrimination between strains within species. By combining results of REP-PCR and ERIC-PCR, five different fingerprint profiles were identified among 17 isolates of Bartonella henselae, but only one profile was identified among the five isolates of Bartonella quintana. Other Bartonella species yielded distinct rep-PCR fingerprints. rep-PCR is a useful technique for identification of Bartonella organisms to the species level and offers the advantage of ease of performance, with only small quantities of cells needed for the whole-cell procedure. This technique also appears to be useful for subtyping B. henselae isolates.

AIDS-Related Opportunistic Infections↗
PubMed1995

Strategy to detect and identify Bartonella species in routine clinical laboratory yields Bartonella henselae from human immunodeficiency virus-positive patient and unique Bartonella strain from his cat.

We wished to develop a cost-effective, rapid strategy to detect and identify Bartonella species in the clinical laboratory and to determine the prevalence of Bartonella infection in the Houston veteran population. Bartonella colonies were identified by colony morphology, Gram stain, RapID ANA, repetitive extragenic palindromic-PCR (REP-PCR) and whole-cell fatty acid (CFA) analysis, and these methods were compared for their usefulness. A new test order for "Rochalimaea culture" (the genus Bartonella was previously known as the genus Rochalimaea) was instituted, and in addition, all blood specimens submitted for fungal culture (obtained in an isolator tube) were processed for Bartonella culture. Over a 16-month period we isolated Bartonella henselae from only 0.4% (2 of 533) of total cultures but from 1% (2 of 204) of human immunodeficiency virus-positive patients. After sufficient growth, identification of the Bartonella isolates to the species level could be obtained in 2 days. The REP-PCR allowed discrimination of all known species, whereas CFA analysis distinguished all except B. henselae and Bartonella quintana. The RapID ANA results failed to differentiate between B. henselae and B. quintana, and results for other species differed by only one or two tests. Blood obtained from a kitten which had been introduced into the household of one patient 2 months before the onset of fever yielded a Bartonella strain which was shown to be different from the strain from the patient and distinct from other Bartonella species by a combination of REP-PCR, CFA, and growth characteristics. Subsequent analysis of the citrate synthase gene sequence showed only an 86% similarity with any of the other known Bartonella species, suggesting that this isolate represents a distinct, previously uncharacterized species of Bartonella.

AIDS-Related Opportunistic Infections↗
PubMed1995

Comparison of agar based media for primary isolation of Helicobacter pylori.

AIMS: To determine the best medium for the primary isolation of Helicobacter pylori. METHODS: Sixty six gastric mucosal biopsy specimens frozen in 1 ml Cysteine Albimi media with 20% glycerol from 22 histologically proven H pylori infected patients were cultured on brain heart infusion agar (BHIA) with 7% fresh whole defibrinated horse blood, egg yolk agar (EYA), Columbia blood agar-cyclodextrin agar (CBA-Cd), and commercial trypticase soy agar (TSA) supplemented with 5% sheep blood. RESULTS: Successful primary isolation of H pylori was 96% with BHIA, 78% with TSA, 64% for EYA, and 32% with CBA-Cd. Colonies appeared earlier on BHIA (4.7 +/- 0.1 days, 5.3 +/- 0.4 days, 5.3 +/- 0.4 days, and 7.1 +/- 0.9 days for BHIA, TSA, EYA, and CBA-Cd) and there were more colonies on BHIA than on CBA-Cd, EYA or TSA (599 +/- 88, 104 +/- 66, 260 +/- 107, and 358 +/- 89, respectively). CONCLUSIONS: Success of a medium for passage of isolates apparently does not reliably predict usefulness for primary isolation. Freshly made BHIA with 7% horse blood medium is recommended for primary isolation. However, the easily obtainable TSA media would be the best alternative for routine clinical laboratories with no access to BHIA.

Agar↗
PubMed1994

Diagnosis of intestinal tuberculosis by polymerase chain reaction on endoscopic biopsy specimens.

It is often difficult to establish the diagnosis of intestinal tuberculosis because of close similarities with other conditions, in particular, Crohn's disease. In the present study, we used the polymerase chain reaction (PCR) assay on endoscopic biopsy specimens obtained from a patient with chronic diarrhea. Positive hybridization was obtained with the Mycobacterium tuberculosis probe and the patient was treated with anti-tuberculous drugs with complete resolution of the endoscopic abnormalities. This study demonstrates that polymerase chain reaction assay can be used on endoscopic biopsy specimens to diagnose intestinal tuberculosis.

Adult↗
PubMed1993

A new Brevibacterium sp. isolated from infected genital hair of patients with white piedra.

A new aerobic gram-positive non-sporeforming bacillus has been isolated from infected genital hair of patients with white piedra in association with Trichosporon beigelii. This species has been characterised morphologically, nutritionally, by DNA base composition, cell-wall analysis and cellular fatty-acid profile on the basis of 14 isolates. The G+C content of DNA is 63.05 mol%. Cell walls possess meso-diaminopimelic acid (Type IV) and the sugars glucose, galactose, xylose and ribose; mycolic acids are not present. The species has a distinct colonial and microscopic morphology, is strongly proteolytic and produces methanethiol. These findings and the cellular fatty-acid profile are compatible with the genus Brevibacterium. A new species is proposed based on the following characters: colonial and microscopic growth and morphology; conditions for rod-to-coccus cycle; ribose utilisation; and tellurite reduction. The type strain has been named Brevibacterium mcbrellneri E2cr (ATCC 49030). The strong proteolytic properties may be the mechanism of pathogenesis.

Anti-Bacterial Agents↗
PubMed1993

Detection of Mycobacterium tuberculosis in clinical samples by two-step polymerase chain reaction and nonisotopic hybridization methods.

Detection of Mycobacterium tuberculosis in clinical specimens by the polymerase chain reaction (PCR) was compared with detection by culture. A 317-bp segment within the M. tuberculosis-specific insertion sequence IS6110 was amplified. The detection limit of the PCR assay for cultured mycobacteria was 50 cells per reaction by ethidium bromide-stained agarose gel electrophoresis and 5 cells per reaction by hybridization with an oligonucleotide probe conjugated with either digoxigenin or alkaline phosphatase (AP). This sensitivity was reduced fivefold in sputum specimens seeded with M. tuberculosis. Seventy-six clinical specimens were amplified and examined by the three detection methods. Both the digoxigenin and AP procedures were found to be more sensitive than agarose gel electrophoresis, but they were occasionally associated with a high background. An additional 308 specimens were examined only by agarose gel electrophoresis and the AP procedure. Of 71 specimens found to contain M. tuberculosis, amplified products were detected from 56 (79%) samples by agarose gel electrophoresis and/or the AP procedure. Of the additional 313 specimens that were culture negative for M. tuberculosis, 19 (6%) had amplified products detectable by agarose gel electrophoresis and/or the AP procedure. Compared with culture, PCR showed sensitivities and specificities of 55 and 98%, respectively, for agarose gel electrophoresis and 74 and 95%, respectively, for the AP procedure. Despite this low sensitivity, a rapid positive PCR result was accurate and clinically useful.

Alkaline Phosphatase↗
PubMed1993

Large-scale use of polymerase chain reaction for detection of Mycobacterium tuberculosis in a routine mycobacteriology laboratory.

We investigated the use of DNA amplification by the polymerase chain reaction reaction (PCR) for detection of Mycobacterium tuberculosis from clinical specimens. Two-thirds of each sample was processed for smear and culture by standard methods, and one-third was submitted for DNA extraction, amplification of a 317-bp segment within the insertion element IS6110, and detection by agarose gel electrophoresis, hybridization, or both. DNA was prepared from over 5,000 samples, with 623 samples being culture positive for acid-fast bacilli. Of 218 specimens that were identified as M. tuberculosis, 181 (85%) were positive by PCR. In the M. tuberculosis culture-positive group, PCR was positive for 136 of 145 (94%) and 45 of 73 (62%) of the fluorochrome smear-positive and -negative specimens, respectively. Of 948 specimens that were either culture positive for mycobacteria other than M. tuberculosis or culture negative, 937 specimens were negative by PCR and 11 (1%) specimens initially appeared to be false positive for M. tuberculosis. The reason for discrepant results varied; some errors were traced to the presence of an inhibitor in the specimen (7.3% in unselected specimens), nucleic acid contamination, low numbers of organisms in the specimen antituberculosis therapy, and possible low-level nonspecific hybridization. In comparison with culture, the sensitivity, specificity, and positive predictive value were 83.5, 99.0, and 94.2%, respectively, for PCR. When PCR was corrected for DNA contamination, the presence of inhibitor, and culture-negative disease, the values became 86.1, 99.7, and 98.4%, respectively. If the results for multiple specimens submitted from the same patient are considered, no patient who had three of more sputum specimens tested would have been misdiagnosed.

DNA, Bacterial↗
PubMed1993

A study of the frequency of recovery of unexpected Giardia lamblia and Cryptosporidium from duodenal aspirates taken during routine upper endoscopy.

UNLABELLED: In this study, our objective was to determine whether the microscopic examination of duodenal aspirates recovered during routine upper endoscopy in unselected, immunocompetent patients, would disclose unsuspected Giardia lamblia or Cryptosporidium. DESIGN: Prospective, cross-sectional study to determine the prevalence of duodenal parasitosis. SETTING: Endoscopy unit in a university hospital. PATIENTS: One hundred and thirty-one unselected, immunocompetent patients undergoing upper endoscopy. MEASUREMENTS: Microscopic examination of duodenal aspirates, obtained during upper endoscopy. No pathogenic or commensal parasite was identified in any patient. CONCLUSIONS: No case of unsuspected parasitosis was found in the present study. On the basis of our findings, we do not recommend routine collection of duodenal aspirates during upper endoscopy.

Adult↗
PubMed1992

Characterization of a novel Rochalimaea species, R. henselae sp. nov., isolated from blood of a febrile, human immunodeficiency virus-positive patient.

Isolation of a Rochalimaea-like organism from a febrile patient infected with human immunodeficiency virus was confirmed. Analysis of 16S rRNA gene sequences, together with polymerase chain reaction and restriction endonuclease length polymorphism analysis of a portion of the citrate synthase gene, demonstrated that the agent is closely related to members of the genus Rochalimaea and that the isolate is genotypically identical to the presumptive etiologic agent of bacillary angiomatosis. However, the same genotypic analyses readily differentiated the new isolate from isolates of other recognized Rochalimaea species as well as other genera of bacteria previously suggested as putative etiologic agents of bacillary angiomatosis and related syndromes. We propose that the novel species be referred to as Rochalimaea henselae sp. now.

Adult↗
PubMed1990

Use of the RapID-ANA system and sodium polyanetholesulfonate disk susceptibility testing in identifying Haemophilus ducreyi.

Haemophilus ducreyi has traditionally been difficult to identify. We have utilized simple test methods to identify 19 fresh isolates obtained during a recent outbreak of chancroid in Houston and six strains of H. ducreyi from other outbreaks. Tests were performed from growth on chocolate agar after 48 h of incubation at 35 degrees C with increased humidity and CO2. All isolates exhibited typical colonial morphology and Gram stain. Isolates were catalase negative and oxidase and nitrate positive (in enriched broth). The RapID NH system failed to identify these strains because of negative reactions with alkaline phosphatase and nitrate reductase. However, by using the RapID-ANA system, all strains were positive for alkaline phosphatase and arginine, glycine, and serine aminopeptidases. Their biochemical profiles were distinct from those obtained with 66 strains representing 13 species similar to H. ducreyi. We also investigated the use of sodium polyanetholesulfonate (SPS) disk susceptibility to identify and differentiate H. ducreyi from other species. All H. ducreyi isolates were susceptible, as evidenced by the presence of a zone of inhibition with an average size of 15 mm around the SPS disk. With the exceptions of Neisseria gonorrhoeae, Gardnerella vaginalis, and Capnocytophaga spp., no other strain showed any evidence of inhibition. The latter three organisms can be easily differentiated from H. ducreyi by various features including reactions in the RapID-ANA. We conclude that, by considering simple growth and biochemical characteristics, SPS susceptibilities, and reactions in RapID-ANA, it is possible for more clinical laboratories to definitively identify this organism.

Bacteriological Techniques↗
PubMed1989

Streptococcus bovis septicemia and large bowel neoplasia.

Streptococcus bovis septicemia is a relatively uncommon entity that is associated with an increased incidence of colonic neoplasms. Three of four patients with S. bovis endocarditis subsequent to septicemia underwent colonoscopy. The fourth patient underwent a barium enema and a proctoscopic examination. Polyps were found in three patients, and adenocarcinoma of the colon in one. Patients with S. bovis endocarditis should be considered at high risk for colonic neoplasms. Screening colonoscopy is recommended for these patients, and follow-up colonoscopy may be warranted.

Adenocarcinoma↗
PubMed1987

Nonpathogenic nematodes in gastrointestinal aspirates obtained during endoscopy.

The gastroenterologist must be alert to diagnosing parasitic diseases when collecting intestinal fluid samples or cytologies during endoscopy. Between 1979 and 1982, we identified nematodes in 10 endoscopic specimens. None were Strongyloides stercoralis. We emphasize the importance and technical aspects of distinguishing nonpathogenic nematodes from Strongyloides.

Colonoscopy↗