Highest Number Of Positive Results Research Articles

Performance of the Roche Elecsys® IGRA SARS-CoV-2 test for the detection and quantification of virus-reactive T cells in COVID-19-vaccinated immunosuppressed patients and healthy subjects

PurposeComparing the performance of commercially available SARS-CoV-2 T-cell immunoassay responses may provide useful information for future observational or intervention studies as well as to their potential customers.MethodWhole blood was collected from a total of 183 subjects fully vaccinated against COVID-19: 55 healthy controls (Group 1), 50 hematological patients (Group 2), 50 chronic kidney disease patients (Group 3), and 28 elderly nursing home residents (Group 4). Samples were tested with the Roche Elecsys® IGRA (Interferon-gamma release assay) SARS-CoV-2 test (Roche Diagnostics, Rotkreuz, Switzerland), the Euroimmun SARS-CoV-2 test (Euroimmun, Lubeck, Germany), the SARS-CoV-2 T Cell Analysis Kit (Miltenyi Biotec, Bergisch Gladbach, Germany), and a flow-cytometry for intracellular cytokine (IFN-γ) staining-based immunoassay (FC-ICS).ResultsOverall, the Roche Elecsys® assay returned the highest number of positive results (151/179; 84.3%), followed by the Euroimmun test (127/183; 69%), and the FC-ICS (135/179; 75%). The Kappa coefficient of agreement was best between IGRAs (0.64). Most discordant results across assays involved patients from Group 2. Overall, IFN-γ concentrations measured by both IGRAs correlated strongly (rho = 0.78; 95% CI 0.71–0.84; P < 0.001) irrespective of the study group. The frequencies of SARS-CoV-2-reactive IFN-γ T cells and IFN-γ concentrations measured by the IGRAs correlated moderately for CD4+ T cells, however, weakly for CD8+ T cells. SARS-CoV-2-experienced participants displayed stronger responses than SARS-CoV-2-naïve when IGRAs, rather than FC-ICS, were used.ConclusionThe SARS-CoV-2 immunoassays evaluated in the present study did not return interchangeable qualitative or quantitative results either in seemingly healthy individuals or in immunosuppressed patients.

Voluntary counselling and testing points (VCTs) in Poland as a complementary and accessible alternative to public healthcare facilities: A comparative analysis of epidemiological data from the VCTs and the National Institute of Public Health NIH-National Research Institute (2015-2022).

Voluntary counselling and testing points (VCTs) offer anonymous and free HIV tests in Poland. They also play an essential role in educational initiatives focused on the prevention and diagnosis of HIV and other sexually transmitted infections. However, no comprehensive data is available that summarizes the results of the work carried out by these VCTs. Therefore, our aim was to conduct a comparative analysis of epidemiological data obtained from VCTs and data reported by the epidemiological surveillance undertaken by the National Institute of Public Health NIH-National Research Institute (NIPH NIH-NRI) covering the period from 2015 to 2022. This retrospective analysis was conducted on data from 258 071 people attending VCTs in Poland in 2015-2022. On average, 32 259 individuals underwent testing each year, with a notable increase in the number of people being tested in November. The average positivity rate was 1.39% (3576/258 071). The Masovian voivodeship conducted the most tests and had the highest number of positive results. The comparative analysis of the frequency of detecting positive results in VCTs and those reported in NIPH NIH-NRI data revealed that, on average, 31.49% (3576/11 356) of positive results in Poland between 2015 and 2022 were identified through tests conducted at VCTs. The positive results identified in VCTs constituted approximately one-third of all results reported by the National Institute of Public Health NIH-National Research Institute, highlighting the importance of VCTs. Moreover, the high availability of testing in the Masovian voivodeship resulted in better detection of HIV. The educational actions performed during European Testing Week increased the number of tests performed in November.

RANGE OF MOTION AFTER BONE BLOCK PROCEDURES FOR SHOULDER INSTABILITY: SYSTEMATIC REVIEW.

to determine the surgical indications for glenoid bone grafting associated with better postoperative ranges of motion. This systematic review was conducted according to PRISMA. The included studies were subdivided according to the criteria used to indicate glenoid bone graft surgery: group for radiological indications only (Group R), group for radiological indications associated with clinical indications (Group R + C), and group for arthroscopic indications (Group A). The extracted and evaluated data were the range of motion of the shoulder. in the electronic search conducted in October 2022, 1567 articles were selected. After applying the inclusion criteria, 14 articles were selected for the systematic review. Regarding the ranges of motion, group A had the highest number of statistically positive results together with group R. Group A showed positive results in elevation parameters, loss of lateral rotation in adduction, and medial rotation in abduction. Group R showed positive results in lateral rotation in adduction and loss of lateral rotation in adduction. On the other hand, Group R + C was the one that presented the highest number of statistically negative results, in the following parameters: elevation, lateral rotation in abduction, loss of lateral rotation in adduction, and medial rotation in abduction. the subgroups presented variable results in the evaluated parameters; however, the groups with arthroscopic and radiological indications showed the highest number of positive results, with the latter group showing the best results regarding lateral rotation. Level of Evidence II, Systematic Reviews.

Using Mass Spectrometry to Assess Disease Status in Treated Multiple Myeloma Patients

Introduction As multiple myeloma (MM) is a bone-marrow based disease, the majority of MRD studies have focused on sensitive bone-marrow based assays. However, to effectively monitor patients this would require sequential invasive procedures at semi-frequent intervals. Sensitive, non-invasive, blood-based assays which can detect the presence of MRD are needed to optimize the management of MM patients. Mass spectrometry (MS) is currently being investigated for its clinical utility at different points in the monoclonal gammopathy pathway, from screening to MRD detection. This study aimed to compare the sensitivity and prognostic utility of MS with other blood-based M-protein detection methods and bone marrow flow cytometry at active disease, best response and MRD timepoints. Methods MS analysis took place at The Binding Site laboratory (The Binding Site, UK) using a pre-commercial quantitative immunoprecipitation (QIP-MS) system. Mass spectra was acquired over an m/z range of 10,000 to 30,000 and analysis was performed by two independent readers. Both transplant eligible and non-transplant chemotherapy only patients were recruited at varying stages of disease and treatments. Median follow up was 18 months. A total of 190 serum samples from 87 individual patients were tested for the presence of M-protein by conventional methods (serum electrophoresis (SPEP), immunofixation (IFE), free light chain (FLC)) and MS. A bone marrow flow cytometry (NGF) assay was performed up to a sensitivity of 10 -6 at pre- and post-transplant for a subset of patients. Disease response was determined according to the IMWG criteria. Results and Discussion When compared to conventional techniques MS showed increased sensitivity over SPEP, IFE and FLC (Table 1). The 1 M-protein detected by SPEP and IFE but not by MS was a post-transplant sample which showed multiple oligoclonal peaks by MS where the original M-protein could not be determined due to not having a diagnostic MS result available. When evaluating the 37 samples from 19 individual patients which were MS positive and IFE negative it was found that 7/19 (37%) patients in this group relapsed during the course of the study. As SPEP, IFE and FLC analyses are often performed together, the utility of using MS alone was compared to an algorithm of SPEP/IFE/FLC tests, where if any of the conventional tests showed abnormality this was considered positive. Using this scenario, MS outperforms the conventional tests in the 190 samples evaluated - with 7 samples being MS positive only and 1 sample (discussed above) being negative by MS but positive by all SPEP/IFE/FLC. As expected, upon analysing the percentage of samples within each IMWG clinical response group which had a positive or negative MS results the percentage of samples with a negative MS result increased with greater depth of response whilst those in partial response or worse had no negative MS samples. However, even in the deepest response group, complete response, 77% of samples were MS positive, showing how MS outperforms the IMWG criteria. Progression free survival (PFS) analysis at the best response time point showed a significant survival advantage of having a MS negative result (p=0.007). When comparing MS against NGF results at bone marrow testing time points agreement at pre-transplant was 88%, with all samples being NGF positive but 22/25 being MS positive and 1 of these 3 discordant patients relapsing during the study. At post-transplant agreement was only 56%. However, evaluating PFS at the post-transplant time point showed that MS status had a significant effect on survival (p=0.02) (Figure 1) where IFE (p=0.44) did not and NGF only approached significance (p=0.051). Conclusion This study aimed to evaluate the viability of using MS to identify M-proteins in treated MM patients. In this study the MS method had a higher number of positive results than any of the conventional methods, and outcompeted the algorithm of SPEP/IFE/FLC in detecting the presence of M-protein. This study has shown a survival benefit of being MS negative at both post-ASCT and best response time points across a range of treatments. There were a high number of patients who were MS positive throughout monitoring and it will be important to determine how such a sensitive method can be used to provide clinically useful information to clinicians to give the best balance between treatment and quality of life for patients.

Towards SDG30: Fighting HCV with a screening program in Lombardy

Abstract Background The third Sustainable Development Goal, specifically in target 3.3, identifies fighting HCV as a priority. It is estimated that in Italy more than 200.000 patients eligible for HCV treatment are not being treated and 20% are not aware of the infection. Lombardy region to promptly identify and treat patients affected, developed a screening program to be offered to all patients born between 1969 and 1989. Healthcare facilities were divided into first and second-level centers and the campaign was designed in 2 phases: at the first level centres patients were screened with blood tests, if positive they were contacted by second level structures to get a diagnostic test and if confirmed positive to start the therapeutic approach. Methods The screening program started in June 2022 and consisted of an opportunistic offer of free testing for serum anti-HCV antibodies in the context of any healthcare performance offered to eligible patients. Data were uploaded and collected on a web database from which second-level centers could obtain data of positive patients to contact them and set appointment for confirmation tests. Results As of 31st December 2022 120,193 individuals underwent HCV screening in 75 screening centers in Lombardy. The mean age was 44 years, 65.2% were females, and 83.7% were tested at outpatient blood collection centers. 604 (0.50%) patients tested positive with mean age 47 (±5). HCV-RNA was detectable in 125 out of 441 (28.3%) anti-HCV positive subjects with available HCV-RNA. Patients with active HCV infection were aged 46 (±6) and were mostly males (56.8%). Overall, the prevalence of active infection in the general population was 0.10% and was higher in elderly. Conclusions The prevalence of active HCV infection in the general population born between 1969 and 1989 in Lombardy was low. Considering the higher prevalence of active infection in the elderly population, an extension of the screening offer to patients born between 1949-70 could be evaluated. Key messages • Prevalence of active HCV infection in screened population is low. • Extending the screening offer may result in higher number of positive results and more people directed towards treatment.

A performance comparison of two (electro) chemiluminescence immunoassays for detection and quantitation of serum anti-spike antibodies according to SARS-CoV-2 vaccination and infections status.

The information provided by SARS-CoV-2 spike (S)-targeting immunoassays can be instrumental in clinical-decision making. We compared the performance of the Elecsys® Anti-SARS-CoV-2 S assay (Roche Diagnostics) and the LIAISON® SARS-CoV-2 TrimericS IgG assay (DiaSorin) using a total of 1176 sera from 797 individuals, of which 286 were from vaccinated-SARS-CoV-2/experienced (Vac-Ex), 581 from vaccinated/naïve (Vac-N), 147 from unvaccinated/experienced (Unvac-Ex), and 162 from unvaccinated/naïve (Unvac-N) individuals. The Roche assay returned a higher number of positive results (907 vs. 790; p = 0.45; overall sensitivity: 89.3% vs. 77.6%). The concordance between results provided by the two immunoassays was higher for sera from Vac-N (ϰ: 0.58; interquartile ranges[IQR]:0.50-0.65) than for sera from Vac-Ex (ϰ: 0.19; IQR:-0.14 to 0.52) or Unvac-Ex (ϰ: 0.18; IQR:0.06-0.30). Discordant results occurred more frequently among sera from Unvac-Ex (34.7%) followed by Vac-N (14.6%) and Vac-Ex (2.7%). Antibody levels quantified by both immunoassays were not significantly different when <250 (p = 0.87) or <1000 BAU/ml (p = 0.13); in contrast, for sera ≥1000 BAU/ml, the Roche assay returned significantly higher values than the DiaSorin assay (p < 0.008). Neutralizing antibody titers (NtAb) were measured in 127 sera from Vac-Ex or Vac-N using a S-pseudotyped virus neutralization assay of Wuhan-Hu-1, Omicron BA.1, and Omicron BA.2. The correlation between antibody levels and NtAb titers was higher for sera from Vac-N than those from Vac-Ex, irrespective of the (sub)variant considered. In conclusion, neither qualitative nor quantitative results returned by both immunoassays are interchangeable. The performance of both assays was found to be greatly influenced by the vaccination and SARS-CoV-2 infection status of individuals.

A Retrospective Population Study of 385 191 Positive Real-Time Reverse Transcription-Polymerase Chain Reaction Tests For SARS-CoV-2 from a Single Laboratory in Katowice, Poland from April 2020 to July 2022.

BACKGROUND This retrospective population study identified 385 191 positive real-time reverse transcription-polymerase chain reaction (RT-PCR) tests for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from a single laboratory in Katowice, Poland, from April 2020 to July 2022. MATERIAL AND METHODS The material was nasopharyngeal, nasopharyngeal swab or bronchial lavage, and bronchoalveolar lavage (BAL) to confirm or exclude SARS-CoV-2 infection with the RT-PCR technique. Personal data are use according to the Provisions on the Protection of Personal Data by the Gyn-Centrum laboratory. RESULTS In 9 months of 2020, the number of SARS-CoV-2 results was 88 986; in 2021, it was 168 439, and in the first 7 months of 2022, it was 12 786. In 2020, the highest number of positive results was recorded in the third quarter (83 094 cases); 2021, in the 1st, 2nd, and 4th quarters (58 712; 37 720; and 71 753 cases, respectively), and in 2022, in the 1st quarter (127 613 cases) of the year. A positive result was observed more often in women and people aged 30-39, followed by those 40-49 years. Patients aged 10-19 years comprised the smallest population of SARS-CoV-2-positive cases. CONCLUSIONS In the Polish population studied, from April 2020 to July 2022, the detection rates of SARS-CoV-2 positivity were significantly higher for women than for men and in the 30-49 age group for both sexes. Also, the infection detection rate of 385 191 out of 1 332 659 patient samples, or 28.9%, supports that the Polish society adhered to public health recommendations for infection control during the COVID-19 pandemic.

Electrophysiological Characteristics of Cervical Spinal Stenosis.

Objective To investigate electrophysiological characteristics of patients with cervical spinal stenosis (CSS) due to cervical disc herniation. Methods A total of 51 patients with CSS diagnosed in our hospital from January 2018 to March 2020 were selected. According to magnetic resonance imaging (MRI), the degree of spinal cord compression was divided into 1-3 grades, namely, group A (MRI grade 1), group B (MRI grade 2), and group C (MRI grade 3), with 17 cases in each group. Subsequently, we analyzed the correlation of the degree of spinal cord compression with the general information, clinical data, and electromyography (EMG) of patients. Results Compared with group A and group B, group C had the longest disease course [(48.06 ± 17.71) months], the lowest JOA score (4.59 ± 2.15), and the highest number of positive results of EMG (EMG: A/B/C, 25/51/77); there were significant differences among the 3 groups. And group C had the higher number of positive cases of both upper and lower limbs in SEP test compared with the other two groups (SEP: A/B/C: 12/18/29: χ2 = 7.559, P = 0.023). According to correlation analysis, MRI grading had no association with gender, age, and spinal canal diameter/volume but was positively correlated with disease course and negatively correlated with JOA score. Conclusion This study primarily verifies that higher MRI grade of CSS is associated with longer disease duration and lower JOA score and EMG. The obtained results secondarily demonstrate the correlation between abnormal neurological status and the MRI grade.

Residuos de antimicrobianos encontrados en aves de corral comercializadas en tiendas minoristas de la zona Metropolitana de Guadalajara, Jalisco

The increased demand to produce large quantities of meat and animal products for human consumption has promoted the indiscriminate use of antimicrobials. The increased use of these substances in the production of poultry, has negative consequences on Public Health due to the fact that the accumulation of antimicrobial residues in the organs and tissues of poultry might reach the consumer. The presence of antimicrobial residues can cause problems of hypersensitivity in humans, or the emergence of antimicrobial resistant pathogens. The purpose of this work was to assess the presence of antimicrobial residues in kidney and muscle tissue of poultry, commercialized in four municipalities of the Metropolitan Area of Guadalajara, Jalisco. The results show that kidney samples had a higher number of positive results compared to muscle tissue. Inhibitors of the folate pathway (sulfamethazine) were the antimicrobials with the highest number of positive results in kidney samples. In contrast, in muscle tissue, β-Lactam (penicillin) were the antimicrobials with the highest number of positive samples. Regarding the analysis of the results by municipalities, it was observed that one of them showed a greater number of positive samples for all the classes of antimicrobials evaluated. This work shows the presence of antimicrobial residues in kidney and muscle tissues of poultry, commercialized in retail sites. Therefore, it is necessary to increase efforts to monitor and control the use of antimicrobial in poultry farms.

Performance of a flow cytometry-based immunoassay for detection of antibodies binding to SARS-CoV-2 spike protein

The performance of a laboratory-developed IgG/IgA flow cytometry-based immunoassay (FCI) using Jurkat T cells stably expressing full-length native S protein was compared against Elecsys electrochemiluminiscent (ECLIA) Anti-SARS-CoV-2 S (Roche Diagnostics, Pleasanton, CA, USA), and Liaison SARS-CoV-2 TrimericS IgG chemiluminiscent assay (CLIA) (Diasorin S.p.a, Saluggia, IT) for detection of SARS-CoV-2-specific antibodies. A total of 225 serum/plasma specimens from 120 acute or convalescent COVID-19 individuals were included. Overall, IgG/IgA-FCI yielded the highest number of positives (n = 179), followed by IgA-FCI (n = 177), Roche ECLIA (n = 175), IgG-FCI (n = 172) and Diasorin CLIA (n = 154). For sera collected early after the onset of symptoms (within 15 days) IgG/IgA-FCI also returned the highest number of positive results (52/72; 72.2%). Positive percent agreement between FCI and compared immunoassays was highest for Roche ECLIA, ranging from 96.1 (IgG/IgA-FCI) to 97.7% (IgG-FCI), whereas negative percent agreement was higher between FCI and Diasosin CLIA, regardless of antibody isotype. The data suggest that FCI may outperform Roche ECLIA and Diasorin CLIA in terms of clinical sensitivity for serological diagnosis of SARS-CoV-2 infection.

Molecular detection of brucellosis in dromedary camels of Qatar by real-time PCR technique

Background and methodologyBrucellosis in livestock causes huge economic loses of developing countries and demonstrates a serious health risks to the patrons of infected dairy goods and meat. Prevention of brucellosis in cattle relies on the trustworthiness of the techniques utilized to detect the causative pathogen. In the current investigation, we described the exploration of the well-known real-time PCR technique based on the Brucella-specific IS711 with different tissue samples of dromedary camels in Qatar. ResultsThe findings of the real-time PCR unveiled the occurrence of Brucella spp in 60 % of lung tissues, 71.42 % of liver tissues, 72 % of spleen tissues, 25 % of kidney tissues, and 42.42 % of placental fluid samples. Among them, the liver tissues and spleen tissues possessed the highest number of positive results, whereas kidney tissues displayed the lowest number of positive results for Brucella spp. ConclusionThe findings of this investigation discloses that PCR technique is a sensitive and specific technique for the identification of Brucella spp. in dromedary camels. Most of the tissues samples showed the presence of Brucella spp. and also we concluded that there is a need for further studies to be conducted in order to identify specific species of Brucella.

Evaluation of rapid diagnostic tests and Enzyme Linked Immunoassay in the detection of malaria in ancient human remains

Malaria has had a significant impact on humans for millennia, yet it has proved difficult to conclusively identify. To-date, there is no absolute way to quantify and confirm the presence of malaria in ancient human remains. Clinically, rapid diagnostic tests (RDTs) are regularly used as a cost-effective and efficient method of identifying the presence of Plasmodium parasite antigens (i.e. malaria) in blood samples. In this study, we evaluate two different immunoassay methods used in the detection of malaria antigens on ancient human skeletal remains, utilising the current generation of RDTs, as well as a highly sensitive histidine rich protein 2 double-site sandwich Enzyme Linked Immunoassay (HRP2-ELISA). Our investigation using RDTs and ELISA immunoassay has produced mixed results, providing new input into, and raising questions regarding, the efficacy and reliability of these immunoassays in the detection of the Plasmodium parasite antigens in ancient bones. Specific protocols to adapt the tests to bone samples must be adhered to, and the sensitivity of the test is crucial to identification. Further optimization of these methods would provide a cost effective avenue of conclusively identifying malaria in the past, and should continue to be pursued. We found that the QDx Malaria Card tests DiaSys produced the highest number of positive results, with 50% of the samples testing positive for malaria with these particular tests.

Epidemiology and drug allergy results in children investigated in allergy unit of a tertiary-care paediatric hospital setting

Background and objectiveDrug Hypersensitivity Reactions (DHRs) are considered adverse effects of medications that resemble allergy symptoms. The reported positive clinical history of pediatric drug reactions is about 10%, however, after allergy investigations, only a small percent is confirmed as hypersensitivity.The aim of this study was to analyze the clinical history, allergy work-up results and sensitization profile of children and adolescents referred to our Allergy Unit for suspected DHRs.MethodsThe study evaluated data related to a group of children with a positive history of drug reactions during a two-year period. The allergy work-up consisted of in vivo and in vitro tests, in accordance with the recommendations of the ENDA/EAACI guidelines.ResultsData from a group of 637 patients [348 M (54.6%); 289 F (45.4%)] were retrospectively analyzed. Beta lactams (BLs) were the most common drugs involved in the reported clinical history, followed by non-steroidal anti-inflammatory drugs (NSAIDs). Severe cutaneous adverse reactions (SCARs) were most frequently observed during BL treatment. The confirmation of BL hypersensitivity was higher for immediate reactions (IRs) [9.4%; 5.1% through positive skin tests (STs) and 5.5% through drug provocation test (DPT)] compared to non-immediate reactions (non-IRs) (8.1%; 2.2% through STs and 6.2% through DPT). A higher number of positive results was obtained for BLs and macrolides when the tests were performed within 12 months after the index reaction (p < 0.05). During DPTs with amoxicillin-clavulanic acid, four hypersensitivity reactions (including one anaphylaxis) occurred despite negative STs.ConclusionOur data demonstrated that only 9.1% of patients resulted in being positive to allergy tests which is in line with the data in literature. An allergy work-up is mandatory for excluding suspected hypersensitivity.

Endometrial histopathology, bacteriology and cytology outcomes in mares with early embryonic death (EED): a field study.

Early embryonic death (EED) is one of the causes of infertility in the mare. We compared endometrial environment in 9 mares with EED and 13 mares in diestrus phase. Cotton swab (CS), cytobrush (CB) and uterine biopsy (B) samples were obtained for the cytological, bacteriological and histopathological examinations. In the first step we compared CS and CB methods to biopsy as a reference method, as B revealed the highest number of positive results in cytological and bacteriological examinations in both groups. In turn, we also compared cytological, bacteriological and histopathological findings between EED and control animals using the B sampling. Although the differences between these groups were not statistically significant (p≥0.05), there was a tendency to a higher prevalence of subclinical endometritis in the control group, than in the EED group (62% vs 22%). In general, positive bacteriological results were similar in both groups (62% vs 55%), whereas positive cytological results were higher in the control group (62% vs 22%; p≥0.05). In histopathological examination in EED mares endometrial degeneration was better expressed (all mares were with grades IIB and III on the Kenney-Doig scale); however, the differences between both groups were not statistically significant (p≥0.05). We could not confirm a clear difference in uterine environment between the two groups. Moreover, the uterine biopsy seemed to be the most reasonable sampling method for diagnosis of endometrial state.

Comparative evaluation of peripheral blood smear preparation techniques as a diagnostic method for hemoparasitosis in dogs (Canis lupus familiaris, Linnaeus, 1758)

Hemoparasitoses are diseases of great importance for animal health, because they cause variable clinical changes, leading to death if not treated. They are caused by intracellular blood parasites like Ehrlichia spp, Anaplasma spp, Babesia spp and Hepatozoon spp, which are of main concern. Hemoparasitoses are called tick born diseases and Rhipicephalus sanguineus is the most involved species, in what concerns to transmission. In addition to several clinical signs, these etiologic agents also cause hematological changes such as anemia. The diagnosis can be made through direct microscopic examination and, also, through more sensitive techniques such as serological and molecular techniques. The goal of this research was to evaluate the most suitable site for peripheral blood collection, used to perform blood smears for direct parasitological examination. Besides, to detect the prevalence of hemoparasites in dogs referred to the Veterinary Hospital of the Federal University of Mato Grosso, situated in Sinop, Mato Grosso, Brazil. From the 66 tested animals, 17 of them (25.75%) were considered positive. Regarding to the performed exams, the buffy coat smears obtained the highest number of positive results, followed by peripheral blood smears and, finally, the ear tip blood smears. Among the most commonly found etiological agents were Anaplasma platys (64.72%), followed by Hepatozoon spp. (17.64%), Babesia spp . (11.76%) and Ehrlichia spp . (5.88%). It is concluded that parasitological direct microscopic examination can be used as a reliable diagnostic method, despite its low sensitivity. Additionally, the observed prevalence of hemoparasites in tested canine population was significant enough to justify an improvement in routine control methods of transmission.

Diagnosis of amphimeriasis by LAMPhimerus assay in human stool samples long-term storage onto filter paper.

Amphimeriasis, a fish-borne zoonotic disease caused by the liver fluke Amphimerus spp., has recently been reported as an emerging disease affecting an indigenous Ameridian group, the Chachi, living in Ecuador. The only method for diagnosing amphimeriasis was the microscopic detection of eggs from the parasite in patients' stool samples with very low sensitivity. Our group developed an ELISA technique for detection of anti-Amphimerus IgG in human sera and a molecular method based on LAMP technology (named LAMPhimerus) for specific and sensitive parasite DNA detection. The LAMPhimerus method showed to be much more sensitive than classical parasitological methods for amphimeriasis diagnosis using human stool samples for analysis. The objective of this work is to demonstrate the feasibility of using dried stool samples on filter paper as source of DNA in combination with the effectiveness of our previously designed LAMPhimerus assay for successfully Amphimerus sp. detection in clinical stool samples. A total of 102 untreated and undiluted stool samples collected from Chachi population were spread as thin layer onto common filter paper for easily transportation to our laboratory and stored at room temperature for one year until DNA extraction. When LAMPhimerus method was applied for Amphimerus sp. DNA detection, a higher number of positive results was detected (61/102; 59.80%) in comparison to parasitological methods (38/102; 37.25%), including 28/61 (45.90%) microscopy-confirmed Amphimerus sp. infections. The diagnostic parameters for the sensitivity and specificity werecalculated for our LAMPhimerus assay, which were 79.17% and 65.98%, respectively. We demonstrate, for the first time, that common filter paper is useful for easy collection and long-term storage of human stool samples for later DNA extraction and molecular analysis of human-parasitic trematode eggs. This simple, economic and easily handling method combined with the specific and sensible LAMPhimerus assay has the potential to beused as an effective molecular large-scale screening test for amphimeriasis-endemic areas.

Control of Hymenolepis nana infection as a measure to improve mouse colony welfare.

After cannibalism had appeared in the reproductive units of a white mouse colony, treatment against confirmed Hymenolepis nana, a tapeworm with zoonotic potential, was performed on 67 mice in the reproductive and nursery units. Faecal droppings were evaluated by flotation and sedimentation methods. The sedimentation method revealed a higher number of positive results before, during and after the treatment, but the flotation method yielded some additional positive cases. In the reproductive unit, H. nana eggs were confirmed in 50% of the tested mice by the flotation and in 70% by the sedimentation method. In the nursery units, H. nana eggs were detected in 10.5% of the tested mice by the flotation and in 24.6% by the sedimentation method. A colony of mice was treated against the tapeworm H. nana with praziquantel and emodepside in doses of 2.574 mg praziquantel/100 g body mass and of 0.642 mg emodepside/100 g body mass. The content of the original pipettes (Profender®) was applied as a spot-on on the back of the neck in the area between the shoulders. The application was repeated three times at 14-day intervals. Seven days after the third therapy no H. nana was found in any of the tested mice in the reproductive or the nursery units. After the treatment, cannibalism was no longer observed. This treatment represented one of the steps aimed at improving animal welfare and preventing potential zoonotic disease. The public health significance of this cestode should receive more attention, especially among people who take care of mice, have them as pets, or feed them to reptiles.

Evaluation of the Direct and Enrichment Iso 10272 Methods for the Detection of Campylobacter Spp. in Chicken Meat

In 2006, the International Organization for Standardization (ISO) published two standard protocols for the detection and enumeration of Campylobacter spp. in foods: the ISO 10272-2(direct method) and the ISO 10272-1 (enrichment method). The aim of this study was to compare the efficiency of these two methods in the detection of Campylobacter spp. and propose a modification in the enrichment method. Compared with the enrichment method, the direct method yielded a higher number of positive results for Campylobacter spp. and, consequently, presented higher sensitivity percentage. The isolation of Campylobacter spp. was more difficult when 10 mL of rinse was used in the enrichment method, which is currently recommended by the ISO protocol. Therefore, different rinse volumes were tested (2.5 and 5 mL). The most efficient recovery of Campylobacter spp. occurred when 2.5 mL of rinse were used in the enrichment method, most likely due to a lower number of microbial contaminants than that present in the 5 or 10 mL rinses. The proposed modification of the enrichment method will contribute to the food analysis by improving the detection of Campylobacter spp. in chicken carcass.

P205 Retesting for genital chlamydia infections in young people is acceptable and important

<h3>Background/introduction</h3> Young persons with a previous history of genital chlamydia are more likely to retest positive at a later date than those that have never tested positive. Current recommendations advise offering a repeat NAAT test to individuals under the age of 25, three months after completing treatment. However, a recall system has yet to be implemented in our clinic. <h3>Objectives</h3> We examined how many young people treated for genital chlamydia voluntarily returned for a further test, to assess whether asymptomatic retesting is both acceptable and worthwhile in this group. We aimed to identify a suitable timeframe in which retesting should be offered. <h3>Methods</h3> Retrospective case note review of individuals under 25 years who tested positive for genital chlamydia in a city wide sexual health service in January 2013. <h3>Results</h3> Of 214 individuals testing positive for genital chlamydia in January 2013, 50% (107/214) retested within 15 months, 29% of which were positive (31/214). Most young people returned 3 to 6 months following their initial diagnosis (37/107), but the highest number of positive results occurred between 7 and 9 months (10/31). Only 8 individuals (7.5%) were retested between 10 and 12 months, though a significant proportion (25%) retested positive. <h3>Conclusion</h3> The high rate of young people returning for chlamydia retesting after a positive diagnosis indicates that retesting is acceptable within this group. The high rate of subsequent positive tests suggests that retesting is important and worthwhile. The ideal timeframe to retest these individuals is 3–12 months following a positive test.

Growth Factor’s Treatment In Musculoskeletal Injuries

The use of platelet-rich plasma (PRP) is increasing as treatment of musculoskeletal system injuries due to sports practice. PURPOSE: To evaluate the security and the therapeutic results of this procedure in different sports injuries. METHODS: 254 patients affected by sports injuries were treated with utrasound-guided PRP injections in the Sports Medicine Clinic of CAR Sant Cugat, Spain from 2010 to 2014. The epidemiological data, the complications recorded and the degree of patient satisfaction (scored from 0 to 10) were retrospectively assessed. Before treatment, all patients signed the informed consent. RESULTS: 254 patients (184 men and 70 women) aged 14-47 years. 165 subjects were elite athletes, 81 soccer players and 57 gymnastics. In 31% of the cases, the treatment was applied to tendon injuries, in 19.5% to synovial pathology and in 14.6% to cartilage pathology. 32.3% of injuries affected the knee, 24.6% the shoulder, and 12.5% the ankle. Mild and self-limiting adverse reactions were recorded only in two patients. One of them manifested pain, oedema and strain sensation; while the other suffered from heavy pain post-infiltration, which disappeared with 1g of paracetamol vo administration. Median satisfaction score was 7 (P25-75 4-8). 48% patients were very satisfied with the treatment. Best results were obtained in tendon injuries (median 8, P25-75 6-10) and in foot injuries (median 9, P25-75 6-10). CONCLUSION: A high number of positive results were obtained from the PRP applications in a group of soft tissue injuries, without important adverse effects. Our results can be taken as a “Proof-of-concept”, although randomized clinic studies are needed in order to demonstrate scientific evidence. Also larger studies might be needed for recommend time between infiltrations, dose, frequency, content, and post-injections rehabilitation techniques.