A Case of Laying Hens Mycosis Caused by Fusarium proliferatum.

In this article, we present the first case report of a chicken mycosis caused by F. proliferatum occurred on a private farm in the Russian Federation. Lesions on the skin of the legs and scallops were reported. The object of this study was samples of feed and pathological material from sick hens-layers. Mycological analysis included determination of the total number of fungi (TNF) and identification and determination of the toxicity and pathogenicity of the isolates. The identification of the isolate was carried out taking into account direct microscopy, morphological features, and the method of molecular genetic analysis. Microscopic fungi of the genus Penicillium and Rhizopus were isolated by mycological analysis of the feed. The test feed was nontoxic. Mycological examination of pathological material (scrapings from the combs and affected legs) identified an isolate of Fusarium proliferatum, which showed toxicity on biological objects (protozoa, rabbits) and pathogenicity (white mice). Dermal application of F. proliferatum suspension was accompanied by reddening of the rabbit skin. Intraperitoneal injection of fungal spores caused mycosis in white mice. Polymerase chain reaction (PCR) made it possible to identify this type of microscopic fungus (F. proliferatum) with high accuracy in the samples under study. The research results allow us to consider F. proliferatum as a cause of poultry disease against the background of predisposing factors in the form of desquamation of the stratum corneum of the skin against the background of immunosuppression and metabolic disorders caused by an imbalance in the diet.

Comparison of a monoclonal with a polyclonal antibody-based enzyme immunoassay stool test in diagnosing Helicobacter pylori infection before and after eradication therapy.

BACKGROUND: Detection of Helicobacter pylori antigen in stool samples has been a subject of controversy. However, it has been included in several clinical guidelines as a recommended non-invasive testing procedure in dyspeptic patients. AIM: To compare a monoclonal enzyme immunoassay for detection of H. pylori stool antigen (Amplified IDEIA HpStAR, DakoCytomation) with a polyclonal enzyme immunoassay (HpSA test, Premier Platinum HpSA, Meridian Diagnostics) in diagnosing infection and in determining H. pylori status after eradication treatment. METHODS: We evaluated stool samples of 198 patients diagnosed with H. pylori infection and of 41 patients without infection. The results of the monoclonal enzyme immunoassay HpStAR were compared with those of the polyclonal enzyme immunoassay HpSA. RESULTS: The sensitivity and specificity of HpStAR were 91.9% and 70.7%, while those of HpSA were 89.4% and 80.5%, respectively. In the 126 patients evaluated 6 weeks after eradication therapy, the overall agreement between urea breath test and HpStAR was 90.5% (P = 0.710) and between urea breath test and HpSA was 76.9% (P = 0.410). CONCLUSIONS: HpStAR is a rapid and easy-to-perform test with similar sensitivity to HpSA in the diagnosis of H. pylori infection, although it had lower specificity. In contrast, HpStAR is more accurate after eradication therapy than HpSA.

Use of a mycobacteriophage-based assay for rapid assessment of susceptibilities of Mycobacterium tuberculosis isolates to isoniazid and influence of resistance level on assay performance.

We standardized and assessed the performance of an in-house microtiter assay for determining the susceptibilities of Mycobacterium tuberculosis clinical isolates to isoniazid based on mycobacteriophage amplification technology. Seventy isolates (43 resistant and 27 sensitive according to the BACTEC 460 radiometric method and MIC determination) were studied. The isoniazid resistance molecular mechanism was previously determined by sequencing the entire katG gene and the mabA-inhA regulatory region. The sensitivity of the mycobacteriophage-based assay in detecting isoniazid resistance was 86.1%, the specificity achieved was 92.6%, and the overall accuracy was 88.6%. In order to assess the possible influence of resistance levels on the mycobacteriophage-based-assay sensitivity, the results were analyzed according to the isoniazid MICs. All the isolates exhibiting high-level resistance (MIC > or = 2 microg/ml) were scored as resistant by the mycobacteriophage-based assay (100% concordance), and 95% showed mutations or deletions in the catalytic domain of the katG gene. In contrast, 26.1% of the low-level-resistance strains (MICs, 0.25 to 1 microg/ml) were misclassified, and 66.7% had alterations in the mabA-inhA regulatory region. The mycobacteriophage-based assay could be used as a rapid method to detect the isoniazid susceptibility pattern, although data from those areas with high rates of low-level-resistance strains should be interpreted with caution. The features of the assay make it suitable for widespread application due to its low technical demand and cost.

Rapid detection of pneumococcal antigen in serum samples for diagnosing pneumococcal pneumonia.

OBJECTIVES: The aim of the study is to assess the usefulness of C polysaccharide and polysaccharide capsular antigen detection by immunochromatography (ICT) and enzyme immunoassay (EIA), respectively, in serum samples for diagnosing pneumococcal pneumonia. METHODS: Adult patients included in the study were classified in the following groups: In group 1 we studied 101 serum samples from patients with pneumonia due to Streptococcus pneumoniae. In 53 cases the pneumonia was bacteremic. The second group contained 113 serum samples from patients with no pneumococcal pneumonia. Group 3 was made up of 40 serum samples from healthy subjects with no clinical or radiological signs of pneumonia. RESULTS: Using ICT, antigen was detected in 50% of patients with pneumococcal pneumonia, in 64.3 and 40.9% of patients with bacteremic and non-bacteremic pneumococcal pneumonia, respectively. Using EIA, antigens were detected in 35.8% of patients with pneumococcal pneumonia, in 45 and 22.2% of patients with bacteremic and non-bacteremic pneumococcal pneumonia, respectively. CONCLUSIONS: In conclusion, the sensitivity of the tests is low. However, in special situations, where obtaining large volume of urine is difficult, they could be a complementary method in the rapid diagnosis of pneumococcal pneumonia.

Efficiency of the incorporation of the hepatitis A vaccine as a combined A+B vaccine to the hepatitis B vaccination programme of preadolescents in schools.

In 1998, the Department of Health of Catalonia (Spain) began universal vaccination of preadolescents against hepatitis A by replacing the simple hepatitis B vaccine with a combined hepatitis A+B vaccine. Economic analyses were made of the two alternative strategies: to continue with the simple hepatitis B vaccination or to replace the simple vaccine with a combined hepatitis A+B vaccine. The analysis was made from the societal perspective and the time horizon considered was 25 years. In the base case, (estimated annual hepatitis A incidence of 15 per 100,000 and incremental price of the hepatitis A+B vaccine over the simple hepatitis B vaccine of 1.98) the net present value of the programme was positive (+533,708) and the benefit-cost ratio was 2.58. If the estimated disease incidence were reduced by half, the programme would still be efficient.

Utility of an in-house mycobacteriophage-based assay for rapid detection of rifampin resistance in Mycobacterium tuberculosis clinical isolates.

A rapid in-house mycobacteriophage-based assay to identify multidrug resistance by detecting the rifampin susceptibility of Mycobacterium tuberculosis in a microtiter plate format was evaluated. The sensitivity, specificity, and overall accuracy of the assay were 100%. This test is rapid to perform and suitable for widespread implementation.

Usefulness of urinary antigen detection by an immunochromatographic test for diagnosis of pneumococcal pneumonia in children.

We evaluated an immunochromatographic assay detecting pneumococcal antigen in urine samples from children diagnosed with pneumococcal pneumonia. The sensitivity and specificity of the immunochromatographic test with nonconcentrated urine (NCU) were 86.7 and 62.9%, respectively; with concentrated urine (CU), they were 100 and 11.7%, respectively. Pneumococcal antigen was also detected in 42.5% of NCU and 87.1% of CU samples from nasopharyngeal carriers. This is a nonspecific test for the diagnosis of pneumococcal pneumonia in children, particularly the very young.

Rapid detection of several mycobacterial species using a polymerase chain reaction reverse hybridisation assay.

The ability of a polymerase-chain-reaction-based technique combined with a reverse hybridisation line-probe assay to detect and identify eight of the most clinically significant mycobacterial species directly from cultures in liquid medium was evaluated. The line-probe assay allows simultaneous identification of eight different mycobacterial species. Ninety-seven mycobacterial strains belonging to 22 different species were evaluated. All strains were previously inoculated into MB/BacT bottles (Organon Teknika, USA). The sensitivity and specificity of the test when applied on positive MB/BacT liquid cultures containing isolates from mycobacterial species included as specific probes on the line-probe assay strip was 100% and 100%, respectively. These results further support the potential clinical usefulness of this technique in the diagnosis of mycobacterial infections.

Detection of Streptococcus pneumoniae antigen by a rapid immunochromatographic assay in urine samples.

STUDY OBJECTIVES: Evaluation of a newly available rapid (15 min) immunochromatographic membrane test (ICT) to detect Streptococcus pneumoniae in urine samples, in order to assess its utility in the diagnosis of bacteremic and nonbacteremic pneumococcal pneumonia. DESIGN: Retrospective study. SETTING: We studied urine samples from 51 patients with bacteremic and nonbacteremic pneumonia due to S pneumoniae diagnosed by blood culture and pneumococcal polysaccharide capsular antigen detection by counterimmunoelectrophoresis in urine samples, 16 patients with probable pneumococcal pneumonia, 71 patients with nonpneumococcal pneumonia, and 16 patients with pneumonia but no pathogen identified. Urine samples were collected and frozen at - 20 degrees C until used. The ICT test was performed following the instructions of the manufacturer. MEASUREMENTS AND RESULTS: S. pneumoniae antigen was detected in 41 of 51 patients with pneumococcal pneumonia (80.4%); results were positive in 23 of 28 bacteremic cases (82.1%) and in 18 of 23 nonbacteremic cases (78.3%). From patients with a diagnosis of presumptive pneumococcal pneumonia, antigen was detected in seven urine samples (43.7%) and also in one case of the 16 patients with pneumonia but no pathogen identified. The specificity of the ICT test was 97.2%. CONCLUSION: The ICT assay is a valuable tool for the diagnosis of pneumococcal pneumonia, especially for the nonbacteremic cases.

Assessment of a new test to detect Legionella urinary antigen for the diagnosis of Legionnaires' Disease.

Given that the rate of mortality by Legionella pneumonia increases in incorrectly treated patients, rapid diagnosis and early antibiotic treatment are needed. We have assessed the performance of a new enzyme immunoassay (EIA) test (Bartels Inc. Trinity Biotech Company, Wicklow, Ireland) to detect Legionella pneumophila antigen in urine comparing it to Binax EIA (Binax, Portland, Maine). We also evaluated the capability of both EIAs to detect extracted soluble antigens of Legionella strains. Using nonconcentrated urine samples (NCU) the sensitivity of Bartels EIA was 74.1% (66/89) and the sensitivity of Binax EIA was 51.7% (46/89). The sensitivity of both EIA tests were 91.5% (54/59) using concentrated urine samples (CU). Specificity of both EIA tests was 100% in NCU and CU. Bartels EIA was able to detect all serogroup L. pneumophila antigens, achieving a higher sensitivity in the case of L. pneumophila serogroup 1 soluble antigen. The new EIA was found to be a useful test for the rapid diagnosis of Legionella pneumonia, being a better alternative to the Binax EIA if NCU is used.

Accuracy of an enzyme immunoassay for the detection of Helicobacter pylori in stool specimens in the diagnosis of infection and posttreatment check-up.

OBJECTIVE: The aim of this study was to assess the reliability of a newly developed enzyme immunoassay for Helicobacter pylori-specific antigen detection in stools (HpSA) compared to other standardized diagnostic techniques such as histology (H), rapid urease test (RUT) and 13C-urea breath test (UBT) to diagnose H. pylori infection and to evaluate its usefulness in determining H. pylori status after treatment. METHODS: One hundred eighty-eight patients referred to our department for upper gastrointestinal endoscopy were included. H. pylori infection was confirmed in all patients by HpSA test in stools, RUT, UBT, and H. Patients were defined as positive for H. pylori if RUT and UBT or H were positive. A total of 142 symptomatic patients received eradication treatment and were reassessed 6 wk after therapy; for 70 of these patients, stool samples were also collected at 24 h and 6 months after finishing eradication treatment. In the posttreatment follow-up, UBT was used as gold standard. RESULTS: The sensitivity of HpSA test for the diagnosis of H. pylori infection using a cut-off value of 0.130 was 89.5% and its specificity 77.8%. This specificity was lower than that obtained with UBT, H, and RUT. In the early follow-up the sensitivity of HpSA test was null. At 6 weeks and at 6 months post-treatment its sensitivity was 70.4% and 50% and its specificity was 81.6% and 79.3%, respectively. CONCLUSIONS: The HpSA stool test, using a cut-off value of 0.130, may be useful for the primary diagnosis of H. pylori infection, with sensitivity similar to that obtained with other standard tests, but with less specificity. HpSA test is not useful for early monitoring of treatment efficacy. At 6 wk and at 6 months posttreatment, HpSA test lacks accuracy as compared to UBT for evaluating the outcome of the eradication treatment.

[The evaluation of a new immunoenzyme analysis for the detection of Helicobacter pylori infection in stool samples]. / Evaluación de un nuevo enzimoinmunoanálisis para la detección de la infección por Helicobacter pylori en muestras fecales.

BACKGROUND: Detection of bacterial antigen in stool specimens (HpSAT) is a new promising tool for diagnosing Helicobacter pylori infection. OBJECTIVE: To evaluate diagnostic accuracy of HpSAT in the diagnosis of Helicobacter pylori infection. PATIENTS AND METHODS: We evaluate the presence of Helicobacter pylori infection by the rapid urease test and the 13C-urea breath test in endoscopic biopsies. Patients who were positive for both tests were considered to have Helicobacter pylori infection. Patients negative for both tests were considered free of infection. The presence of Helicobacter pylori infection was also determined in stool specimens by means of HpSAT. RESULTS: The sensitivity of HpSAT was 92.8%, the specificity 92.3%, the positive predictive value 98.1% and the negative predictive value 75%. CONCLUSIONS: HpSAT is a reliable tool for diagnosing Helicobacter pylori infection.

Evaluation of a rapid immunochromatographic assay for the detection of Legionella antigen in urine samples.

A new immunochromatographic membrane assay for detecting Legionella pneumophila serogroup 1 antigen in urine samples (Binax Now Legionella Urinary Antigen Test; Binax, USA) was evaluated. Its sensitivity, specificity and level of agreement with the Binax enzyme immunoassay were compared using nonconcentrated and concentrated urine samples. The overall agreement between the two tests was 98.1%; the specificity of both was 100%. The sensitivity of the immunochromatographic assay was 55.5% in nonconcentrated urine and 97.2% in concentrated urine in comparison with the enzyme immunoassay, using concentrated urine as the reference test. This immunochromatographic assay screens successfully for Legionella pneumophila serogroup 1 soluble antigen in concentrated urine samples.

Evaluation of Meridian ImmunoCard Mycoplasma test for the detection of Mycoplasma pneumoniae-specific IgM in paediatric patients.

The Meridian ImmunoCard Mycoplasma kit, a 10-min card-based enzyme-linked immunosorbent assay (ELISA) designed to detect immunoglobulin M (IgM) antibodies to Mycoplasma pneumoniae was evaluated. We compared the ImmunoCard with the Fujirebio Serodia Myco II particle agglutination test, as well as with the complement fixation (CF) test to detect M. pneumoniae antibodies in paediatric patients. The ImmunoCard test and Serodia Myco II test agreed in 93.95%, and ImmunoCard test and CF test agreed in 83.51% of the 182 specimens tested. Nine specimens gave negative particle agglutination titres in the acute phase sample, and 28 specimens gave negative CF titres in the acute phase sample, although in the ImmunoCard test they were positive. These results may indicate that the ImmunoCard assay detects lower IgM levels of antibodies than the Serodia Myco II and CF test. The ImmunoCard appears to be a good screening assay test for M. pneumoniae IgM in children in whom M. pneumoniae IgM is found frequently.

Comparison of the Binax Legionella urinary antigen enzyme immunoassay (EIA) with the Biotest Legionella Urin antigen EIA for detection of Legionella antigen in both concentrated and nonconcentrated urine samples.

We evaluated a newly commercial enzyme immunoassay (EIA) (Biotest Legionella Urin Antigen EIA; Biotest AG, Dreieich, Germany) for detection of antigens of all Legionella pneumophila serogroups with a relatively wide spectrum of cross-reactivity as well as antigens of other Legionella spp. by comparing its sensitivity and specificity with those of an EIA for detection of L. pneumophila serogroup 1 antigen (Legionella urinary antigen EIA; Binax, Portland, Maine). Both tests were performed with both concentrated and nonconcentrated urine samples. We also evaluated the capabilities of both EIAs to detect extracted soluble antigens of American Type Culture Collection (ATCC) Legionella strains (L. pneumophila serogroups 1 to 14, L. bozemanii, and L. longbeachae). The sensitivity of the Biotest EIA was 66.66% in nonconcentrated urine and 86.66% in concentrated urine. The sensitivity of the Binax EIA was 63.76% and 88.88% in nonconcentrated and concentrated urine, respectively. The specificity was 100% in nonconcentrated and concentrated urine for both assays. The Binax EIA and Biotest EIA detected extracted soluble antigens of L. pneumophila serogroups 1 to 14 and L. bozemanii ATCC strains. The cross-reactions observed with the Binax EIA were probably due to common epitopes directly related to lipopolysaccharide. Further studies are required to determine the usefulness of the Binax EIA for detection of urinary antigens from Legionella species and serogroups other than L. pneumophila serogroup 1. The Biotest EIA proved to be as rapid, sensitive, and specific as the Binax EIA for the diagnosis of legionellosis. Concentration of antigen present in urine increased the sensitivities of both techniques with no reduction in specificity.

Importance of sexual transmission of hepatitis C virus in seropositive pregnant women: a case-control study.

The mode of hepatitis C virus (HCV) transmission in patients who deny parenteral exposure is still not understood. Seroprevalence studies of anti-HCV in sexually promiscuous populations and in spouses of infected patients have given contradictory results. We investigated the role of sexual transmission of HCV in a case-control study of risk factors for infection in a series of 43 anti-HCV positive pregnant women and 172 matched controls (4 for each case). In the univariate analysis, the following factors were associated significantly with anti-HCV seropositivity: low social class, unmarried, history of abortion, wounds which were sutured, tattooes, sharing toiletries with the partner, sexual contact outside the partnership without condom use, blood transfusion, and intravenous drug abuse, but only the last 3 factors remained significantly associated with HCV infection in multiple logistic regression analysis. The relative risk of HCV infection increased according to the increased number of sexual partners. Thus sexual transmission must be considered a possible mode of infection in HCV infected persons without parenteral exposures.

[The efficacy of the prevaccination detection of anti-HAV in hepatitis A vaccination programs]. / Eficiencia de la detección prevacunal de anti-VHA en los programas de vacunación antihepatitis A.

BACKGROUND: The convenience of carrying out prevaccination detection studies of hepatitis A virus (HAV) markers depends on the relative costs of the detection and vaccination, as well as the prevalence of susceptible subjects in each population group to be vaccinated. The aim of this study was to analyze the efficacy of the systematic prevaccination detection of anti-HAV antibodies in Catalonia, Spain. METHODS: The following formula was applied: Unit cost of detection + (1-X) x Unit cost of vaccination of anti-HAV negative subjects = group vaccination cost, with X being the threshold of prevalence of marker under which detection no longer remains efficient. This prevalence was compared with the findings of a seroepidemiologic survey carried out in international travellers and food handlers. RESULTS: The unit cost of the detection of anti-HAV was calculated as 2,733 pesetas, and the unit cost of the vaccination as 9,963 pesetas obtaining a prevalence anti-HAV threshold of 27%. This prevalence corresponds to travellers under the age of 30 years and food handlers under the age of 25 years. CONCLUSIONS: The systematic detection of anti-HAV is only recommended in population groups in which prevalences higher than 27% may be expected. The vaccination is more efficient without a previous marker study under this threshold. According to this study, direct vaccination of food handlers under the age of 25 years (born after 1969) is recommended as is that of international travellers under the age of 30 years (born after 1964). In those over the age of these collectives, the prevaccination study is more efficient.