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Biomedical subjects

D Vaira

Publications and source records attributed to D Vaira.

At least 55 records · Page 3Linked to original sources
PubMed2000

An assessment of the long-term preservation of the DNA of a bacterial pathogen in ethanol-preserved archival material.

To examine the potential for DNA recovery from spirit-preserved medical material, a set of specimens from the Hunterian Collection of the Royal College of Surgeons was investigated. Using a range of DNA extraction techniques and the PCR, no replicable positive amplifications were made from this material of either human or Helicobacter DNA. Experiments with modern stomach biopsies of H. pylori-positive patients suggest that the bacterial DNA is typically present in a much lower concentration (10(3)-fold) than that of the host. The potential for recovery of this organism from spirit specimens is therefore low. The absence of DNA in this material is probably due to several factors, chiefly the incomplete fixation of the specimen by the ethanol storage fluid. Studies such as this demonstrate the need for a good understanding of specimen history when working with archival material.

Archives↗
PubMed2000

Image analysis method to assess adhesion of Helicobacter pylori to gastric epithelium using confocal laser scanning microscopy.

We have used confocal scanning microscopy of FITC-labelled bacteria to assess binding of Helicobacter pylori to stomach sections and to assess the effect of inhibitors on binding to the Lewis antigens. We have quantified the binding using an image manipulation package that is readily available on the web. Our results demonstrate heterogeneity of binding of Helicobacter pylori to tissue sections and that binding can be inhibited using synthetic Lewis B oligosaccharide.

Bacterial Adhesion↗
PubMed2000

Accurate diagnosis of Helicobacter pylori. Stool tests.

From the limited data available, it seems that the H. pylori stool assay represents a highly accurate diagnostic tool to detect H. pylori infection before and shortly after therapy. As a test that is noninvasive, accurate, simple, and cost-effective, the H. pylori stool assay has the potential to become the preferred diagnostic tool in many different clinical settings from epidemiologic studies to pediatric investigations, from pre-endoscopic screening policies to posttherapy monitoring.

Antigens, Bacterial↗
PubMed2000

Helicobacter pylori seroconversion in asymptomatic blood donors: a five-year follow-up.

Several techniques have been developed to diagnose Helicobacter pylori infection and two noninvasive methods are available: carbon 13-urea breath test (UBT) and serology. Measurement of IgG serum antibodies by enzyme-linked immunosorbent assay (ELISA) is a reliable and inexpensive method for detection of infection. The aim of this study was to assess the seroconversion by different techniques after five to eight years. In 1990, 588 of 1,010 asymptomatic donors were found to be seronegative by ELISA, based on an H. pylori whole-cell suspension lysate (sensitivity and specificity: 92% and 97%). In 1995 serum samples from 418 of 588 seronegative donors were collected and retested using the same antigen. 411 of 418 samples were frankly negative, and 7 donors were found to be seroconverted. This group of seven sera represents the object of the study. They were retested by ELISA and western blotting using a different antigen (NCTC). To standardize our techniques, sera from 43 H. pylori positive and 47 H. pylori negative patients according to culture, histology, urease test, and UBT were used. The cutoff for ELISA-NCTC was 0.53 AI (absorbance index) (mean value + 2 SD), and for western blotting was negativity for CagA or <10 bands (sensitivity and specificity: 95% and 96%; 98% and 81% for ELISA and western blotting respectively). According to the results obtained in 1990 and 1995, seven donors were found to be seroconverted by ELISA using sonicated antigen; in five the seroconversion was confirmed by ELISA using NCTC antigen and in two there was concordance with WB. Four of the seven donors were contacted and asked to undergo UBT and a further serum sample was drawn to be reassessed in 1998. A seroconversion was found in all four donors by ELISA, while WB and UBT confirmed the seroconversion in only three of four donors. In conclusion the in-house ELISA used performed well compared to other theoretically better serologic assays and confirmed the low seroconversion rate for H. pylori infection in adult populations living in developed countries.

Adult↗
PubMed2000

Isotope ratio mass spectrometry (IRMS) versus laser-assisted ratio analyzer (LARA): a comparative study using two doses of.

This study was carried out to compare the measurements and the diagnostic accuracy of the traditional expensive IRMS and the new economical LARA system using two doses of [13C]urea + two different test meals in patients undergoing upper gastrointestinal endoscopy, both before and after anti-Helicobacter treatment. A total of 354 dyspeptic patients underwent endoscopy with gastric biopsies to diagnose H. pylori infection by CLO-test and histology. No patients had taken antibiotics, bismuth, or antisecretory drugs in the 4 weeks before testing. After overnight fasting, breath samples were collected simultaneously in both plastic and glass tubes at baseline and at 30 and 60 min after urea ingestion. In 237 patients 100 mg [13C]urea + Ensure and in 117 patients 75 mg [13C]urea + citric acid were given. The test was also performed with the two urea dosages and meals in 67 and 64 infected patients, respectively, four weeks after anti-Helicobacter therapy. H. pylori was considered eradicated when both biopsy-based tests were negative. A delta value >5 per thousand was considered positive. Breath samples with insufficient CO2 levels at both 30 and 60 min were excluded from final analysis (N = 37 in pre- and N = 8 in posttreatment). There was excellent agreement between overall delta values of the two machines with both [13C]urea 100 mg + Ensure and [13C]urea 75 mg + citric acid. The 95% CI of the difference against the mean was wider with the former (mean -1.3, +6.3, and -9.4) than with the latter urea dosage and test meal (mean -1.2, +5.2 and -8.1). LARA and IRMS were equally effective (P = NS) in distinguishing infected from uninfected patients before therapy using both doses of [13C]urea and test meals (sensitivity ranged from 95% to 99% and specificity from 95% to 97%). This good performance was maintained in the posttreatment phase (sensitivity ranged from 90% to 100% and specificity from 90% to 97%), without any statistical difference among the various combinations (P = NS). The LARA system is a valid alternative to IRMS in the diagnosis of H. pylori infection. Both machines provide highly reliable results after 30 min, so that the 60 min sample can be avoided. The dose of 75 mg + citric acid suffices to ensure an accurate UBT. The test performed with both devices and [13C]urea dosages is very effective also for posttherapy evaluation of H. pylori status.

Adult↗
PubMed2000

Review article:invasive and non-invasive tests for Helicobacter pylori infection.

There are two general ways in which a diagnosis of infection by Helicobacter pylori can be made: by using either an invasive or non-invasive procedure. The invasive procedures involve an endoscopy and biopsy. A biopsy is essential because often the mucosa may appear macroscopically normal but nevertheless be inflamed. A biopsy is obtained by histological examination, culture, polymerase chain reaction or detection of the presence of urease activity in biopsy material. The non-invasive tests that can be used to diagnose the infection are serology, detection of labelled metabolic products of urea hydrolysis in the breath (13CO2, 14CO2), the urine or the blood, and detection of H, pylori antigen in a stool specimen. At present no single test can be relied upon to detect definitely colonization by H. pylori, and a combination of two is recommended if this is feasible. The choice of the test to be used is not straightforward and may vary according to the clinical condition and local expertise.

Breath Tests↗
PubMed2000

Blinded, multicenter quality control study for the quantification of human immunodeficiency virus type 1 RNA in plasma by the Belgian AIDS reference laboratories.

OBJECTIVE: In order to evaluate the interlaboratory variation of HIV-1 RNA measurements in plasma, the Belgian AIDS reference laboratories organized a blinded multicenter quality control study. METHODS: Atest panel of coded spiked HIV-1 plasma samples reflecting the dynamic range of the assay was composed and distributed. The HIV-1 RNA concentration of these samples was determined by the eight Belgian AIDS reference laboratories by means of the Amplicor HIV-1 Monitor version 1.5 assay. RESULTS: Analysis of the results demonstrated that there was little interlaboratory variation for the high concentration range (4.0-5.7 log10 copies/mL), never exceeding 0.2 log10 copies/mL. However the standard deviation for the low concentration range (2.6-3.9 log10 copies/mL) reached up to 0.22 log10 copies/mL. CONCLUSIONS: Since interlaboratory variability never reached 0.5 log10 copies/mL and each of the laboratories was able to detect four-fold differences in plasma HIV-1 RNA levels, the Amplicor assay can be used in multicenter studies without a centralized analysis of samples. Furthermore, this well-characterized proficiency panel of spiked plasma samples could be used as a standard in the study of interassay comparisons.

Belgium↗
PubMed2000

Noninvasive antigen-based assay for assessing Helicobacter pylori eradication: a European multicenter study. The European Helicobacter pylori HpSA Study Group.

OBJECTIVE: In a recently published multicenter study involving 501 patients undergoing esophagogastroduodenoscopy (EGD) throughout Europe, we showed the high accuracy of a recently developed simple test (HpSA) to detect Helicobacter pylori (H. pylori) antigens in stools of untreated patients. The aim of this study was to assess the diagnostic usefulness of HpSA compared with 13C UBT shortly after H. pylori eradication treatment. METHODS: Of the 501 patients enrolled in the validation study, 279 were found to be H. pylori-positive. These patients were given H. pylori eradicating regimen and asked to return for follow-up EGD with biopsies, 13C UBT and HpSA testing 4 wk after therapy. Follow-up results were available for 235 patients. Of these, 162 consented to all testing and 73 consented only to 13C UBT and HpSA testing. We assessed sensitivity and specificity of both HpSA and 13C UBT compared with biopsy-based methods in the 162 patients, who accepted follow-up EGD. We also assessed sensitivity and specificity of HpSA compared with 13C UBT, arbitrarily chosen as the gold standard, in the whole population of 235 patients. RESULTS: Sensitivity and specificity in 162 patients who consented to a second EGD were 93.8% (CI: 85.4-100%) and 96.9% (CI: 93.9-99.9%) for HpSA, and 90.6% (CI: 80.5-100%) and 99.2% (CI: 97.7-100%) for UBT. Using EGD-based methods as the gold standard, 130 of the 162 treated patients' H. pylori infection were eradicated (125 HpSA-negative, one borderline, and four false-positive; 129 13C UBT-negative, one false-positive), and 32 remained H. pylori-infected (30 HpSA-positive, two false-negative, 29 13C UBT-positive, three false negative). The overall eradication rate was 80.2%. The sensitivity and specificity of HpSA relative to UBT as the gold standard in the overall population (n = 235) were 95.6% (CI: 89.6-100%) and 94.7% (CI: 91.5-97.9%), respectively. CONCLUSIONS: HpSA has proven to be a useful method in posttreatment eradication testing for H. pylori. Its ease of use, speed, and noninvasive nature make HpSA testing an ideal method for post-treatment monitoring where a second EGD may not be justified.

Adult↗
PubMed1999

Diagnosis of Helicobacter pylori infection with a new non-invasive antigen-based assay. HpSA European study group.

BACKGROUND: Helicobacter pylori is a common human pathogen implicated in certain gastrointestinal diseases. In the search for new non-invasive techniques to diagnose H. pylori infection, we evaluated an EIA for H. pylori antigen in stool (HpSA). METHODS: In a prospective multicentre study, stool specimens from 501 patients (276 men, 225 women; age range 17-88 years, mean 52) undergoing gastroscopy in 11 centres throughout Europe were tested with HpSA and the carbon-13-urea breath test. At endoscopy, four biopsy samples were taken for histology (haematoxylin and eosin) and H. pylori detection (giemsa in both antrum and corpus, culture and rapid urease test). Patients were defined as positive for H. pylori if histology (antrum, corpus, or both) and urease test were positive, or if culture was positive. Patients classified as having H. pylori infection received an eradication regimen; 107 were reassessed 4 weeks after therapy. FINDINGS: Of 272 patients with H. pylori infection by the predefined criteria, 256 were positive by HpSA (sensitivity 94.1% [95% CI 90.6-96.6]). Of 219 patients without infection, 201 were negative by HpSA (specificity 91.8% [87.3-95.1]). INTERPRETATION: The stool assay was a reliable and easy-to-use tool for diagnosis of H. pylori infection. The test was accurate even shortly after treatment.

Adolescent↗
PubMed1999

Anti-CagA reactivity in Helicobacter pylori-negative subjects: a comparison of three different methods.

Emerging evidence suggests that infection by CagA-positive Helicobacter pylori strains is related to the development of more serious gastroduodenal diseases, thus conferring to the determination of anti-CagA antibodies a relevant clinical significance in serological screenings. The detection of anti-CagA positivity in sera negative for anti-H. pylori antibodies raises the question of whether this apparently nonsense result is merely due to a false positive reaction. To address this issue, we compared three different methods for the detection of anti-CagA antibodies. In all, 272 selected sera from patients with precisely defined H. pylori status (positive or negative concordance of five tests, ie, histology by Giemsa in both antrum and corpus, rapid urease test, culture, [13C]urea breath test, IgG ELISA) were tested for anti-CagA reactivity by three different techniques (western immunoblotting, ELISA, and recombinant immunoblotting assay). In order to assess the sensibility and specificity of each tests, we considered as "true" anti-CagA positive sera those with two out of three positive results. Sera from 70% of H. pylori-positive patients and 10% from H. pylori-negative patients turned out to be "true" positives for anti-CagA antibodies. The three methods showed similar excellent results, in terms of both sensitivity and specificity, always over 93%. It is confirmed that a proportion of patients with a negative conventional serology against H. pylori possess anti-CagA antibodies in their sera. In this paper we demonstrate that it can happen even in patients without any biological signs of actual H. pylori infection. The possibility that this can be due to a false positive laboratory result is very likely ruled out by the accuracy of the three methods used. The clinical management of these patients needs further study on larger series.

Adult↗
PubMed1999

High prevalence of Helicobacter pylori infection in shepherds.

It has been suggested that Helicobacter pylori infection may, in some instances, be a zoonosis. The aim of this study was to evaluate the prevalence of H. pylori infection in Sardinian shepherds and their families in relation to exposure to sheep and sheep dogs. Sardinian shepherds and a control group of blood donors completed detailed questionnaires regarding demographics, childhood and current economic status, and the presence of symptoms related to the upper gastrointestinal tract. H. pylori status was determined by a sensitive ELISA for anti-H. pylori IgG and by western blot for anti-CagA IgG. A subgroup of shepherds had upper gastrointestinal endoscopy with biopsy to assess the severity of the gastritis. H. pylori infection in Sardinian shepherds approached 100% and was positively related to animal contact (98% of shepherds, 73% of family members without regular direct animal contact compared to 43% of blood donors) (P < 0.001). Importantly, the family members shared the same childhood with the shepherds but choose different careers (e.g., teachers, nurses, business) and did not have regular contact with sheep. In conclusion, the prevalence of H. pylori infection in Sardinian shepherds is among the highest in the world and is associated with direct contact with sheep and sheep dogs. These results suggest that the cycle of H. pylori infection might, in certain circumstances, include phases in the environment, animals (sheep or dogs) and human beings.

Adolescent↗
PubMed1999

Laser assisted ratio analyser 13C-urea breath testing, for the detection of H. pylori: A prospective diagnostic European multicentre study.

BACKGROUND: Novel technology based on laser optogalvanic spectroscopy called the LARA (Laser Assisted Ratio Analyser) system was developed to measure 12C/13C ratios in breath samples using stable 13C isotopes, to detect Helicobacter pylori infection. AIM: To determine the sensitivity and specificity of the 13C-LARA-urea breath test in the detection of H. pylori infection in a prospective European multicentre trial; FDA-and EMEA-approved. METHODS: Consecutive dyspeptic patients underwent diagnostic gastroscopy with biopsies for culture and histopathology, to detect H. pylori infection (gold standard). Subsequently, the LARA-urea breath test was performed using either a system without a cold trap (part I) or a system with a cold trap (part II). In both instances baseline, 30-min and 60-min breath samples were collected. The optimum cut-off level for 12C/13C ratios was determined by Receiver Operator Characteristics analysis. RESULTS: In part I, 544 out of 604 patients were evaluable (low CO2: 47; withdrawn: 13). 284 out of 544 patients (52%) were H. pylori-positive according to the gold standard. The sensitivity of the LARA-urea breath test was 95% and the specificity 94%. In part II, 257 out of 272 were evaluable (low CO2: 14; withdrawn: 1). Sensitivity and specificity were 93% and 96%, respectively. CONCLUSION: The LARA-technology represents an accurate and non-invasive testing system for the detection of H. pylori infection. Its major advantages are the use of stable 13C isotope, the high throughput of samples and the easy means of collecting, storing and transporting the samples, thus making the system convenient to both patient and clinician.

Adolescent↗