Search PubMed⌕ Search

Biomedical subjects

V J Gill

Publications and source records attributed to V J Gill.

At least 37 records · Page 2Linked to original sources

Cytomegalovirus infection in children with human immunodeficiency virus infection.

OBJECTIVES: To determine retrospectively the prevalence of positive cytomegalovirus (CMV) cultures in pediatric patients with human immunodeficiency virus infection. METHODS: We reviewed the records of 273 children with human immunodeficiency virus infection referred to the Pediatric Branch of the National Cancer Institute for whom CMV cultures were performed between January, 1991, and October, 1994. RESULTS: Of this group 189 patients (69%) had negative CMV cultures and 84 (31%) had positive cultures. The prevalence of CMV-related disease was 9% for the entire group, including 4 (2.1%) patients with negative CMV cultures. Among the 84 patients with positive CMV cultures, 21 (25%) had evidence of CMV disease. Patients with positive CMV cultures had a statistically significant decrease in survival in the presence of severe immunocompromise defined as an age-corrected CD4 count of < 21%. Nine of 35 (26%) autopsies performed demonstrated evidence of CMV disease, including 7 patients with disseminated CMV disease. CONCLUSIONS: Although CMV disease appears to be less frequent in children than adults, CMV infection still contributes significantly to morbidity and mortality in this population, especially when combined with severe immunosuppression.

AIDS-Related Opportunistic Infections↗

Lack of clinical utility of bronchoalveolar lavage cultures for cytomegalovirus in HIV infection.

This study assessed the presence of cytomegalovirus (CMV) in bronchoalveolar lavage (BAL) in three subpopulations of HIV-infected patients and correlated its presence with clinical status during 3 mo of follow-up. Nineteen asymptomatic volunteers, six patients with CMV retinitis, and 46 patients with acute pulmonary symptoms underwent BAL and were assessed for CMV by cytopathology, conventional shell vial cultures, and antigen detection. Transbronchial biopsies were also obtained when possible and evaluated for histopathologic changes of CMV. All patients were followed for approximately 3 mo. Cytomegalovirus was detected in BAL in nine of 19 (47%) asymptomatic volunteers, in all six patients with CMV retinitis, and in 33 of 46 (72%) patients with pulmonary symptoms. Only one symptomatic patient with a positive CMV BAL culture developed clinically significant CMV pulmonary disease; this patient developed disseminated CMV and died. The only other death occurred in a patient with CMV retinitis who developed staphylococcal bacteremia. None of the asymptomatic volunteers or patients with CMV retinitis developed evidence of CMV pneumonia or any other organ disease with CMV. Cytomegalovirus is frequently detected in BAL from HIV-infected patients regardless of their pulmonary symptoms and its presence does not clinically predict significant pulmonary morbidity or mortality in 3 mo of follow-up.

AIDS-Related Opportunistic Infections↗

The laboratory evaluation of opportunistic pulmonary infections.

The patient population at risk for opportunistic pulmonary infections has increased during the last decade. The spectrum of organisms causing opportunistic infections has also grown. With an ever broader list of potential diagnosis, a specific diagnosis of the cause of pulmonary disease becomes more important. Recent microbiologic advances have helped to facilitate the laboratory diagnosis of some of these agents. Immunoassays are available for the detection of antigen in nasopharyngeal secretions (respiratory syncytial virus, influenza) in serum (Cryptococcus species), and in urine (Legionella or Histoplasma species). Rapid-culture techniques are available for the culture and detection of various viruses, including cytomegalovirus. Molecular probes can now assist in the rapid identification of Mycobacterium tuberculosis and some fungi. In the near future, polymerase chain reaction-based techniques may assist in the detection of Pneumocystis carinii and Legionella, Chlamydia, Mycoplasma, and Mycobacteria species. An expeditious evaluation of pulmonary disease requires an understanding of the differential diagnosis of likely causes of pulmonary disease in specific immunosuppressed patient populations, an understanding of the most appropriate specimens to process for these diagnoses, and an understanding of the limitations (sensitivity and specificity) of these diagnostic tests. An understanding of the most appropriate specimens and tests in a given institution should allow for early, relatively specific treatment of many potentially life-threatening infections.

Clinical Laboratory Techniques↗

Clinical evaluation of the BacT/Alert and isolator aerobic blood culture systems.

The BacT/Alert (BTA) (Organon Teknika, Durham, NC) and Isolator 10 (ISO) (Wampole Laboratories, Cranbury, NJ) blood culture systems were evaluated for their ability to detect aerobic and facultatively anaerobic microorganisms in blood of adult patients. For each culture 8 mL of blood was inoculated into both the aerobic standard BTA bottle and the ISO tube. Of 7,259 paired culture sets, 1,168 organisms were recovered, and 667 (57.1%) of these were considered clinically significant. This represented 540 clinically significant positive cultures from 266 patients. Of the significant isolates, 410 were recovered by both systems, 108 by BTA only and 149 by ISO only (P <.025). Overall, the BTA detected 77.7% of the significant isolates, whereas ISO detected 83.8%. The ISO recovered significantly more isolates of Staphylococcus aureus (P = .0001), coagulase-negative Staphylococcus spp (P <.01), and non-Enterobacteriaceae gram-negative rod species (P <.0025), whereas the BTA detected significantly more isolates of Streptococcus spp (P <.0025). Growth of S aureus (P <.0025), Enterococcus spp (P <.0025), and Streptococcus spp (P <.0075) was detected earlier by the BTA when laboratory coverage was available during the first shift only (7:30 AM to 4:00 PM), and additionally of Enterobacteriaceae (P <.0005) and other gram-negative rod species (P <.0001) if coverage was extended to 12:00 AM. Yeasts were detected more rapidly by the ISO (P <.0025). The ISO contamination rate (5.9%) was six times that of the BTA. Taking into account its ability to rapidly detect most organisms, its automated and thus labor-saving features, and the minimal contamination rate associated with its use, the BTA appears to be a reliable alternative to the ISO as a blood culturing system, although improvement in detection of staphylococci and non-Enterobacteriaceae gram-negative rods would be desirable.

Adult↗

Severe Burkholderia (Pseudomonas) gladioli infection in chronic granulomatous disease: report of two successfully treated cases.

Chronic granulomatous disease (CGD) is characterized by a defect in phagocytic cells that leads to recurrent superficial and deep pyogenic infections. Burkholderia (Pseudomonas) gladioli is a gram-negative bacillus in the pseudomallei group of pseudomonads that is known primarily as a plant pathogen. We report two cases of pneumonia, one accompanied by septicemia, caused by B. gladioli in patients with CGD and their successful treatment with antibiotics. We believe these represent the first reports of human disease caused by this organism. We conclude that B. gladioli should be considered a potential pathogen in patients with CGD.

Adult↗

PCR amplification of rRNA intergenic spacer regions as a method for epidemiologic typing of Clostridium difficile.

From January to March 1993, a suspected outbreak of antibiotic-associated diarrhea occurred on a pediatric oncology ward of the Clinical Center Hospital at the National Institutes of Health. Isolates of Clostridium difficile obtained from six patients implicated in this outbreak were typed by both PCR amplification of rRNA intergenic spacer regions (PCR ribotyping) and restriction endonuclease analysis of genomic DNA. Comparable results were obtained with both methods; five of the six patients were infected with the same strain of C. difficile. Subsequent analysis of 102 C. difficile isolates obtained from symptomatic patients throughout the Clinical Center revealed the existence of 41 distinct and reproducible PCR ribotypes. These data suggest that PCR ribotyping provides a discriminatory, reproducible, and simple alternative to conventional molecular approaches for typing strains of C. difficile.

Bacterial Typing Techniques↗

Comparison of pigment production and motility tests with PCR for reliable identification of intrinsically vancomycin-resistant enterococci.

Forty-eight clinical isolates identified as either Enterococcus faecium or Enterococcus faecalis with the MicroScan system (Dade International, MicroScan Inc., West Sacramento, Calif.) were further characterized by two supplementary biochemical tests (pigment production and motility). Twenty isolates (42%), all initially identified as E. faecium, were motile. Of these 20, 8 isolates (17%) produced yellow pigment and were identified as Enterococcus casseliflavus and the remaining 12 (25%) were nonpigmented and were identified as Enterococcus gallinarum. Identical identification results were obtained when PCR amplification of regions of the vanC gene was used as a technique for differentiating these organisms. The results of this study indicate that motility and pigment production tests together with commercial test systems are sufficient for reliable identifications of E. faecium, E. casseliflavus, and E. gallinarum.

Bacteriological Techniques↗

Development and evaluation of a rapid and simple procedure for detection of Pneumocystis carinii by PCR.

We report the development of a simplified PCR-based assay for the detection of Pneumocystis carinii DNA in clinical specimens. The adoption of a rapid DNA extraction procedure and the introduction of a type of enzyme-linked immunosorbent assay for PCR product detection enabled this procedure to be carried out in a single working day in a clinical microbiology laboratory. The PCR assay was prospectively compared with an immunofluorescent-antibody (FA) staining method for the detection of P. carinii in induced sputum and bronchoalveolar lavage (BAL) specimens. The results of the study showed that, for induced sputum specimens, FA staining had a sensitivity of 78% (32 of 41 specimens) and a specificity of 100% (166 of 166 specimens); PCR was 100% (41 of 41 specimens) sensitive and 98% (162 of 166 specimens) specific. For BAL specimens, FA staining was 100% sensitive (21 of 21 specimens) and 100% specific (133 of 133 specimens), and PCR had a sensitivity of 100% (21 of 21 specimens) and a specificity of 99% (132 of 133 specimens). These results strongly suggest that use of our PCR-based assay could effect clinically useful improvements in the sensitivity of induced sputum specimens for the detection of P. carinii.

Bronchoalveolar Lavage Fluid↗

Improved enrichment broth for cultivation of fastidious organisms.

An enrichment broth developed in our laboratory, fastidious broth (FB), was compared with two commercially available broth media, supplemented thioglycolate broth and enriched eugonic broth. FB supported the growth of a number of organisms that were not cultivatable in either of the other two media, including Corynebacterium jeikeium, Haemophilus influenzae, Neisseria gonorrhoeae, and Streptococcus pneumoniae. In addition, for several organisms that were able to grow in all three broths, including Neisseria meningitidis, Nocardia asteroides, and Actinomyces spp., both the time of incubation and the starting inoculum necessary to enable detection of growth were decreased significantly by using FB.

Bacteria↗

Identification of clinical isolates of nondiphtherial Corynebacterium species and their antibiotic susceptibility patterns.

Starting in 1982, our laboratory has performed species identification of coryneform bacteria isolated from blood cultures, intravenous (i.v.) catheter tips and sites, urines with high colony counts, and other potentially significant cultures, using predefined criteria. Of 283 isolates identified, Corynebacterium jeikeium was the most common (47%), followed by CDC group G2 (12%) and C. minutissimum (8%). Blood cultures and i.v. catheter-related sources were the most frequent sources (58% of total). Certain species or groups, like CDC group G2, were most frequently isolated from blood or i.v. catheter sites. CDC group G2 showed a progression to greater multiple antibiotic resistance during this 9-year period. Occasional multiresistant strains of other species were also encountered. By in vitro testing, we note vancomycin remains the most active agent against corynebacterialike organisms, and is the most reliable antibiotic to use while awaiting susceptibility testing results.

Blood↗

Detection of Chlamydia pneumoniae by polymerase chain reaction-enzyme immunoassay in an immunocompromised population.

Chlamydia pneumoniae has now been associated with pneumonia, bronchitis, pharyngitis, acute chest syndrome of sickle cell disease, and asthma. Because of the difficulty of primary isolation and tissue-culture adaptation of this organism, we used a previously developed polymerase chain reaction-enzyme immunoassay (PCR-EIA) to screen 132 culture-negative bronchoalveolar lavage (BAL) specimens from 108 immunocompromised patients (34% of whom were positive for human immunodeficiency virus) and 7 healthy volunteers. Thirteen specimens (9.8%) from 12 immunocompromised patients (11.1%) gave a positive result; one patient had two positive specimens obtained 3 days apart. No healthy volunteer had a PCR-EIA-positive BAL specimen. Twelve (11.1%) of the immunocompromised patients also had diagnostic levels of antibody. Four patients had positive results in both PCR-EIA and serological tests. Thus 20 (18.5%) of the 108 patients had laboratory evidence of C. pneumoniae infection. These data indicate that diagnosis of acute infection with C. pneumoniae can be established more rapidly and reliably by PCR-EIA than by culture or serology, particularly among immunocompromised patients, in whom serological changes in response to infection are relatively undependable. With an infection rate of 11.1% according to PCR-EIA, C. pneumoniae should be considered in the evaluation and treatment of pneumonia in immunocompromised patients.

AIDS-Related Opportunistic Infections↗

Esophageal disease in AIDS is associated with pathologic processes rather than mucosal human immunodeficiency virus type 1.

Twenty-five patients with AIDS and esophageal symptoms were evaluated for the presence of esophageal disease and human immunodeficiency virus type 1 (HIV-1) in the esophageal mucosa. A single infectious process caused by Candida albicans, cytomegalovirus, herpes simplex virus, varicella-zoster virus, or Mycobacterium avium-intracellulare complex or a single noninfectious process caused by Kaposi's sarcoma or reflux esophagitis was identified in 20 patients. Two processes were identified in 5 patients. HIV-1 mRNA was detected by in situ hybridization in mononuclear cells in esophageal lamina propria in 36% of patients. The presence of HIV-1 in the esophageal mucosa was not associated with a specific esophageal symptom, mucosal inflammation or ulceration, Kaposi's sarcoma, specific opportunistic infection, or response of the infection(s) to therapy. Esophageal disease in patients with AIDS appears to be associated with specific pathologic processes rather than the presence of HIV-1 in esophageal mucosa.

AIDS-Related Opportunistic Infections↗

Detection of microsporidian spores in clinical samples by indirect fluorescent-antibody assay using whole-cell antisera to Encephalitozoon cuniculi and Encephalitozoon hellem.

Three polyclonal mouse antisera, to Encephalitozoon cuniculi, Nosema algerae, and Nosema corneum, and two polyclonal rabbit antisera, to E. cuniculi and Encephalitozoon hellem, were used in an indirect fluorescent-antibody assay (IFA) with Enterocytozoon bieneusi, E. cuniculi, and Encephalitgozoon. hellem spores (spores of the last two were taken from culture). Enterocytozoon bieneusi cannot be cultured. By IFA, antisera to E. cuniculi and E. hellem reacted strongly and equally with each other's spores. The mouse antisera reacted strongly with the homologous species, but for these there was segmental and particulate or "dot" staining of heterologous microsporidian spores, indicating cross-reactions with more selected antigens. In fecal samples, cross-reactions with both mouse and rabbit antisera were sometimes seen with different yeast species, with species of streptococci, and species of gram-negative rods. There were no cross-reactions to staphylococci. Enterocytozoon bieneusi was easily identified in duodenal and colonic biopsies, duodenal and colonic fluids, and feces of symptomatic AIDS patients by IFA. In a study of 12 AIDS patients with diarrhea, the new IFA identified microsporidia in all of 11 fecal samples, three colon fluids, six duodenal fluids, and three duodenal biopsy touch preparations. Although the fecal sample of 1 of the 12 was negative, the patient's duodenal fluid contained microsporidian spores by IFA.

Animals↗

Rapid method for presumptive identification of Corynebacterium jeikeium.

Corynebacterium jeikeium causes systemic infections, particularly in immunocompromised hosts. A minitube assay has been developed for the presumptive identification of C. jeikeium. With our rapid sucrose-urea test and conventional biochemical tests, sixty isolates of gram-positive, catalase-positive bacilli were identified in our laboratory. Results indicated that our assay has a sensitivity of 100% and a specificity of 90%.

Algorithms↗

Improved diagnosis of Pneumocystis carinii infection by polymerase chain reaction on induced sputum and blood.

Detection of Pneumocystis carinii by the polymerase chain reaction (PCR) may facilitate non-invasive diagnosis of P carinii pneumonia and study of its epidemiology. We have compared the sensitivity and specificity of two PCR methods with those of conventional staining for detection of P carinii in induced sputum, bronchoalveolar lavage fluid (BAL), and blood. Of 71 sputum samples, 17 were from patients with microbiologically confirmed P carinii pneumonia. A nested PCR method correctly identified the presence of P carinii in all 17 (100% sensitive, 95% confidence interval [CI] 81-100%) and found no organisms in 50 of 54 microbiologically negative samples (93% specific, 95% CI 82-98%). PCR with a single primer pair was 71% sensitive (44-90%) and 94% specific (85-99%). The sensitivity of conventional staining methods (direct and indirect fluorescence antibody and toluidine-blue-O tests) was significantly less (38-53%) than that of nested PCR (p less than 0.05). In BAL, neither PCR method was significantly better than the conventional staining methods. P carinii was detected in BAL or sputum from 10 immunocompromised patients without microbiological evidence of P carinii pneumonia, which suggests that symptom-free carriers or subclinical infection can exist. P carinii was detected by nested PCR in blood from 2 of 3 patients with disseminated pneumocystosis but in only 1 of 11 patients with P carinii infection restricted to the lungs. Nested PCR on induced sputum is more sensitive than conventional staining methods for the diagnosis of P carinii pneumonia and provides a non-invasive method of detecting disseminated disease.

Animals↗

Pneumocystis carinii and specific fungi have a common epitope, identified by a monoclonal antibody.

Because Pneumocystis carinii may be related to fungi, we evaluated the reactivities of monoclonal antibodies raised against P. carinii with a variety of fungi. Fifty-two fungi and six protozoa were evaluated by immunofluorescence. One of three monoclonal antibodies (MAbs) tested (MAb 7D7) reacted with 15 fungi but no protozoa. Saccharomyces cerevisiae showed the strongest reactivity by immunofluorescence. The reactive antigen was characterized for four fungi by the immunoblot technique. In all cases the antigen that was reactive with MAb 7D7 was larger than the P. carinii antigens that reacted with 7D7. In further studies with P. carinii, Aspergillus species, and S. cerevisiae, we found that MAb 7D7 reacted with a carbohydrate component in all organisms. The presence of an epitope that is common to P. carinii and a number of fungi further supports the fungal nature of P. carinii.

Antibodies, Monoclonal↗

Diagnosis of Pneumocystis carinii pneumonia by multiple lobe, site-directed bronchoalveolar lavage with immunofluorescent monoclonal antibody staining in human immunodeficiency virus-infected patients receiving aerosolized pentamidine chemoprophylaxis.

The yields of both induced sputum examination and bronchoalveolar lavage (BAL) have been reported to be decreased for breakthrough episodes of Pneumocystis carinii pneumonia in human immunodeficiency virus-infected patients receiving aerosolized pentamidine chemoprophylaxis. This study assessed whether the yield of a single middle or lower lobe BAL could be increased by the utilization of two techniques: (1) indirect immunofluorescent staining with a combination of two murine monoclonal anti-Pneumocystis antibodies in addition to routine toluidine blue O and cytopathologic staining, and (2) the performance of multiple lobe, site-directed BAL (i.e., both upper lobe and middle or lower lobe lavage, including the lobe with the greatest radiographic abnormality). Results of 252 fiberoptic bronchoscopies performed at the National Institutes of Health and the Los Angeles County-University of Southern California Medical Center were analyzed. P. carinii pneumonia was documented in 21 episodes in patients who did not receive prior anti-Pneumocystis chemoprophylaxis and in 41 episodes in patients who received aerosolized pentamidine. Monoclonal antibody staining and multiple lobe, site-directed BAL resulted in similar diagnostic yields for P. carinii in the nonprophylaxis (100%) and aerosolized pentamidine (98%) groups. If BAL had been performed without monoclonal antibody staining and multiple lobe, site-directed lavage, then the yield would have decreased to 95% in the nonprophylaxis group and to 80% in the aerosolized pentamidine group.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS-Related Opportunistic Infections↗