Rapid Detection Kit Research Articles

Evaluation of the applicability of immunochromatography to the identification of live plague vaccines and the tularaemia allergen (Tularin)

The regulatory standards require that the identification of live plague vaccines and the liquid tularaemia allergen (Tularin) should be performed by immunofluorescence. A major drawback of the recommended method is its labour intensive nature. However, immunochromatography represents an alternative method that offers a number of advantages, including rapid testing and easy result interpretation. The aim of the study was to assess the applicability of immunochromatography to the identification of live plague vaccines and the liquid tularaemia allergen (Tularin).Materials and methods. The authors performed identification tests using samples of the pharmacopoeia standard for live plague vaccines, three commercial batches of a live plague vaccine, and two batches of the liquid tularaemia allergen (Tularin). These samples were tested using immunochromatographic assay (ICA) reagent kits for rapid detection and identification of Yersinia pestis (ICA System for Y. pestis) and Francisella tularensis (ICA System for F. tularensis) manufactured by the State Scientific Center for Applied Microbiology and Biotechnology.Results. The findings show that immunochromatography is an effective, rapid, and species-specific method to confirm the presence of Y. pestis in a sample of a live plague vaccine or F. tularensis in a sample of the liquid tularaemia allergen (Tularin). To perform identification tests by immunochromatography, the authors recommend diluting live plague vaccine samples to a concentration of 109 bacterial cells/mL and using undiluted samples of the liquid tularaemia allergen (Tularin).Conclusions. The study results may support the inclusion of ICA into the regulatory standards for live plague vaccines and the liquid tularaemia allergen (Tularin) as an alternative identification method.

Regeneration of T cells from human-induced pluripotent stem cells for CAR-T cell medicated immunotherapy.

Background: Chimeric antigen receptor (CAR) T cell treatment involves in vitro production of T cells from patient blood with synthetic receptors specific to a cancer antigen. They circumvent the major histocompatibility complex to recognize the tumor antigen, reducing hematologic malignancy remission rates by 80%. Considering the efficacy of CAR-T treatment, the present work aimed at generating functional clusters of differentiation (CD)8 + T cells from human induced pluripotent stem cells (hiPSC) and to generate hiPS-CAR-T cells with high antigen-specific cytotoxicity. Methods: The Alkaline phosphatase assay and MycoEasy rapid mycoplasma detection kit was implemented for detection of hiPSCs and mycoplasma, respectively. The CD34+ HSPCs were harvested in AggreWellTM 400 using a 37-micron reversible strainer. Likewise, the lymphoid progenitor and CD4+CD8+ DP T cells were also harvested. The Cell Counting Kit-8 (CCK-8) assay was used to mark cytotoxicity and ELISA was used to detect IFN-γ secretion. Further, flow cytometry and transwell chambers were used to assess cell cycle, and migration and invasion. Finally, the in vivo antitumor effects of the CAR-T cells were evaluated using experimental animals (mice). Results: Results revealed that a serum-free, feeder layer-free differentiation system significantly yielded hiPSC-based T cell immunotherapy with interleukin-2, interleukin-15, and activators at the differentiation stage to promote the maturation of these cells into human induced pluripotent stem (hiPS)-T cells. The infection of hiPSCs with the CD19 CAR lentivirus resulted in the production of the hiPSC-CAR-T cells. We validated the function of hiPS-CAR-T cells in vivo and in vitro experimentation which revealed no significant differences in cell morphology and function between hiPSC-derived hiPS-CAR-T cells and peripheral blood-derived CAR-T cells. Conclusion: This study developed a culture method that is efficient and clinically useful to make functional CD8+ T cells from hiPSC and to get hiPS-CAR-T cells with high antigen-specific cytotoxicity that are not very different from CAR T cells found in peripheral blood. As a result, our findings may open the way for the clinical use of hiPSC to create functional CD8+ T and hiPS-CAR-T cells cells for use in cell-based cancer therapy.

즉석섭취식품에 존재하는 Salmonella spp.와 Listeria monocytogenes의 검출을 위한 SureTectTM와 표현형 및 유전자형 방법의 비교

The objective of this study is to compare and assess the effectiveness of real-time polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), and the selective agar plate method for the detection of Salmonella spp. and Listeria monocytogenes in ready-to-eat (RTE) foods. In RTE foods, the detection performance of the three methods (RT-PCR [SureTectTM kit and PowerChekTM kit], LAMP [3M MDS], selective agar) were similar at 0-10, 10-50, 50-100, and 100- CFU/mL of Salmonella spp. and L. monocytogenes. We found that with RT-PCR, the Ct value of salad was significantly higher (p<0.05) than other RTE foods, indicating that fiber plays a critical role as an obstacle to the rapid detection of Salmonella spp. However, the Ct value displayed a mixed pattern according to the inoculation level of L. monocytogenes. The use of rapid detection kits and machines mostly depends on the user’s choice, with accuracy, ease of use, and economy being the primary considerations. As an RT-PCR kit, SureTectTM and PowerChekTM showed high accuracy in detecting Salmonella spp. and L. monocytogenes in RTE foods, showing that they can replace the existing RT-PCR kits available. Additionally, LAMP also showed excellent detection performance, suggesting that it has the potential to be used as a food safety management tool.

Submerged leaves of live indoor foliage plants adsorb H1N1 influenza virus from suspension

ABSTRACT Control of hazardous indoor particles using plants has attracted interest due to the increasing worldwide air pollution and spread of pandemic-causing viruses. However, the interaction between human pathogenic viruses (HPVs) and live plants has not been examined largely due to issues in detecting tiny amounts of infectious viruses in a carrier (such as an aerosol) and the lack of suitable examination methods. In this study, as a novel evaluation method, the effect of submerged leaves of live plants on HPVs in water was examined, using the H1N1 influenza virus as a model. Selected plant foliage of a live plant was immersed in a small bag containing HPV water suspension. In an initial screening test, the activities of 20 different plant species on the virus suspension were evaluated using a rapid virus detection kit. Ten plant species had the capability to decrease virus concentrations in the water suspension within 72 h. Among the experimental plant species, Epipremnum aureum showed the highest virus decreasing characteristics when examined using both the kit and quantitative real time polymerase chain reaction. The capacity of immersed leaf of live E. aureum to decrease viral content was enhanced when the plant-containing pot was electrically grounded to the earth (approximately 70% decrease in virus concentration). The foliage sample analysis showed that virus adsorption to the plant foliage surface could be the major reason for the decrease in the suspension. These results suggest that the proposed method can be applied to select plants to further investigate plant–HPV interactions.

An update on lateral flow immunoassay for the rapid detection of SARS-CoV-2 antibodies.

Over the last three years, after the outbreak of the COVID-19 pandemic, an unprecedented number of novel diagnostic tests have been developed. Assays to evaluate the immune response to SARS-CoV-2 have been widely considered as part of the control strategy. The lateral flow immunoassay (LFIA), to detect both IgM and IgG against SARS-CoV-2, has been widely studied as a point-of-care (POC) test. Compared to laboratory tests, LFIAs are faster, cheaper and user-friendly, thus available also in areas with low economic resources. Soon after the onset of the pandemic, numerous kits for rapid antibody detection were put on the market with an emergency use authorization. However, since then, scientists have tried to better define the accuracy of these tests and their usefulness in different contexts. In fact, while during the first phase of the pandemic LFIAs for antibody detection were auxiliary to molecular tests for the diagnosis of COVID-19, successively these tests became a tool of seroprevalence surveillance to address infection control policies. When in 2021 a massive vaccination campaign was implemented worldwide, the interest in LFIA reemerged due to the need to establish the extent and the longevity of immunization in the vaccinated population and to establish priorities to guide health policies in low-income countries with limited access to vaccines. Here, we summarize the accuracy, the advantages and limits of LFIAs as POC tests for antibody detection, highlighting the efforts that have been made to improve this technology over the last few years.

A paper stick test kit for rapid detection of total microbial contaminants in fresh milk

The way to determine the total microbial contaminants in fresh milk is by the Total Plate Count (TPC) method. The TPC method requires 48 hours, high analytical skills, and high analytical costs. The aim of this research was to design a rapid detection tool for microbial contaminants in fresh milk called Paper Stick Test Kit (PSTK), which is fast, cheap, and easy. The stages of research designing PSTK are: (a) making PSTK, (b) testing the optimal time for using PSTK, and (c) making standard color maps. The results of this research are: (a) the best PSTK is made from cardboard with color indicators made from a mixture of bromothymol blue 0.3 g/250 ml solvent and NaOH 0.01 N; drying PSTK is best done in an oven with a temperature of 100oC, for 20 minutes; (b) the optimal time of application of PSTK in fresh milk is 20 seconds; and c) standard color maps are based on the correlation between microbial contaminant levels in fresh milk and color levels in PSTK dipped in microbial contaminated fresh milk.

Development of a universal kit for rapid detection of aggressive chemicals, rocket fuel components and uranium components and uranium compounds in various environments

The materials of the article are devoted to the issues of ecology and safety at enterprises and railway transport. The urgency of developing a universal kit for rapid detection of aggressive chemicals, rocket fuel components and uranium compounds in various environments is due to the fact that the presence on the surface and in water can lead to poisoning and chemical burns of working personnel and the public. It is also caused by the need to make managerial decisions when eliminating the consequences of accidents at food and related industries. The article discusses the methods of rapid detection of aggressive chemicals and suggests scientific, methodological and technological approaches to the development of a universal set of tools for group rapid detection of aggressive chemicals and uranium compounds in water and on the surfaces of objects using aerosol devices with original indicator formulations. The place of research is Moscow and the Moscow region

Seroepidemiological Study of Dengue Virus Infection Suspected Cases in Burkina Faso

Dengue fever is a mosquito-borne disease caused by four related but antigenically distinct dengue viruses. In the last half-century, the prevalence of dengue fever has increased dramatically (up to 30%) worldwide, especially in Sub-Saharan Africa. In Burkina Faso, the last dengue epidemic occurred in 2016, and the epidemiological situation is still poorly documented at the national level. Therefore, the present study aimed to conduct a sentinel seroepidemiology of suspected dengue virus infections from 2017 to 2020 at Charles de Gaulle University Pediatric Hospital Center of Ouagadougou. This investigation was designed to be a descriptive and analytical study. Patients with suspected cases of dengue fever were recruited from January 2017 to December 2020 during general medicine consultation and referred to Charles de Gaulle University Pediatric Hospital Center of Ouagadougou for the serological diagnosis of dengue. Venous blood was collected in dry or EDTA tubes and tested for DENV NS1 antigen, anti-dengue IgM, and anti-dengue IgG using SD Bioline Dengue Duo Rapid Detection Kit (Standard Diagnostic Inc., Korea). A total of 3400 blood samples from clinically suspected dengue cases were analyzed, of which 1784 (52.5%) were males, and 1616 (47.5%) were females. Among the 3400 patients included in the study, 661 (19.4%) were tested at least positive for NS1 antigen, anti-dengue IgM or anti-IgG. Among them, individuals positive for IgG suggesting past dengue virus infection were found in more than a third of 262 (7.7%) of the cases. Approximately, 80.0% (2705/3400) dengue suspected cases and 85.5% (341/399) early or primary dengue infections were recorded in the last four months of the study (September to December 2020), with a peak in mid-October and mid-November. There was no statistically significant difference in seroprevalence between males and females (p = 0.7), but an increasing seropositivity trend with age, from 11.6% for the patient under 5 years group to 39.7% for 30 years and over age group were noted. This study puts in evidence a considerable level of transmission of dengue viruses in Burkina Faso and provides baseline seroprevalence data. The recurrent outbreaks of dengue infection in multiple geographical areas need comprehensive surveillance and a diagnostic system to identify the actual burden and pinpoint the risk factors. A larger study is, therefore, needed to determine the actual prevalence of dengue in Burkina Faso and map the serotypes.

Clinico-Epidemiological Profile and Utility of Diagnostic Techniques in Immunocompetent Children with Cryptosporidium Diarrhoea

Purpose: To assess the Clinical and Epidemiological profile of children 6 months to 5 years having Cryptosporidium diarrhoea and to compare the diagnostic accuracy of rapid diagnostic test (RDT) against modified acid-fast stain technique to detect Cryptosporidium in stool. Materials and Methods: Details of demography, presenting complaints, and detailed examination of children presenting with acute or persistent diarrhoea were recorded in a predesigned proforma. Two Stool samples were collected and examined same day by rapid antigen test and microscopy for presence of antigen or oocyst of cryptosporidium. Results: Out of the total of 280 samples tested, 40 (14.3%) samples were positive for Cryptosporidium. The prevalence of Cryptosporidium in children presenting with diarrhoea in our study came out to be 14.3%. Fever, vomiting and pain abdomen were the most common presenting complaints. Epidemiological factors like providing clean water, proper feeding and hygiene, and avoidance of overcrowding can help in reducing the burden of Cryptosporidium diarrhoea. Persistent diarrhoea was more common among cryptosporidium positive cases. Rapid detection kits enhance the detection rate and are sensitive as well as specific. It could be utilised as an aid in diagnosis of Cryptosporidium diarrhoea. Conclusion: The study shows that cryptosporidial diarrhoea is not uncommon in immunocompetent children. Improvement in personal hygiene, feeding practices and access to safe and potable water can help in reducing burden of this disease. Antigen detection kits aid in early diagnosis. Keywords: Paediatric cryptosporidiosis, Persistent diarrhoea, Sanitation, RDT in diarrhoea, Abdominal pain

Sulfur-fumigated ginger identification via brightness information and voting mechanism

Abstract As a food consumed worldwide, ginger is often sulfur-fumigated. Sulfur-fumigated ginger is harmful to health. However, traditional methods to detect sulfur-fumigated ginger are expensive and unpractical for the general public. In this paper, we present an efficient and convenient identification method based on image processing. First, rapid detection kits were employed to mark three levels of sulfur-fumigated gingers, and the RGB images of the gingers of each sulfur-fumigated level are collected. Second, the brightness and texture features were extracted from the images. Three machine learning methods, Support Vector Machine, Back Propagation Neural Network and Random Forest, were applied to establish prediction models. Third, the accuracy of each model was calculated and different weights were assigned for different models. Finally, models with different weights determined whether the ginger was sulfur-fumigated or non-sulfur-fumigated, and then the results were summarized to establish the final identification model. The experimental results show that the proposed method is robust. When the training set occupies 90%, the prediction accuracy is up to 100%. When the training set only occupies 10%, the accuracy remains high at 80%. Meanwhile, the proposed method is more competitive than other methods in terms of accuracy.

Determination of Paddy Soil Ammonia Nitrogen Using Rapid Detection Kit Coupled with Microplate Reader.

Indophenol blue colorimetry has been widely used for determining soil ammonia nitrogen, but this method has some disadvantages, such as complex reagent preparation, high toxicity, and long colorimetric time. Hence, we aimed to develop a rapid soil ammonia nitrogen determination method using a rapid detection kit. In order to select a suitable extractant, different concentrations of KCl and NaCl solutions were used to extract soil. The ammonia nitrogen content in different types of soils was determined using a rapid detection kit (purchased from Zhejiang Luheng Environmental Technology Limited Company) coupled with a microplate reader. The kit method was compared with the traditional indophenol blue colorimetry method. The results showed no significant difference between the 1 mol·L-1 KCl extraction kit method and indophenol blue colorimetry (p > 0.05). The linearity of the working curve was smooth, the linear detection range was 0.0-2.00 mg·L-1, the average relative standard deviation was 7.00% (n = 5), the standard addition recovery rate was 89.31-118.23%, and the detection limit were was 0.074 mg·L-1. We concluded that the 1 mol·L-1 KCl extraction kit method can be applied to determine the ammonia nitrogen content of paddy soil with different chemical properties. The 1 mol·L-1 KCl extraction kit method has the advantage over indophenol blue colorimetry due to its simple reagent preparation, convenient operation, and shorter detection time (the coloring and colorimetric time for 96 samples was only 30 min using the kit method coupled with a microplate reader). Hence, it has the potential for application due to its rapid determination of soil samples in large quantities.

A portable kit for rapid detection of bromadiolone in human blood and urine via surface-enhanced raman scattering coupled with salt-induced liquid-liquid phase separation

Bromadiolone is the most frequently detected anticoagulant rodenticide in human biofluids, resulting in increased cases of poisoning. Here, we present a portable kit that could realize rapid fingerprint detection of DQ in human biofluids. This strategy was based on surface-enhanced Raman scattering (SERS) and salt-induced liquid-liquid phase separation (SILLPS) using a gold/silver core-shell nanoparticle (Au@Ag NPs) substrate. The kit included a portable Raman spectrometer and five tubes that contained Au@Ag NPs colloid, KI solution, saturated NaCl solution, acetonitrile, and Mg(NO3)2 solution. This strategy did not require pretreatment for urine samples. For blood samples, a 2-min SILLPS method was proposed with satisfactory recovery, providing unique insights into efficient pretreatment of complex biomatrix. The detection limits for urine and blood were calculated to be 3.21 ppb and 6.86 ppb, respectively. A clear trend could be observed between signal intensity and logarithmic concentration of bromadiolone ranging from 0 to 2 ppm with high reproducibility. Such a portable kit based on SILLPS-SERS can rapidly provide fingerprint information of poison. This simple-yet-practical “Pretreat & Detect” SERS kit may open up new opportunities for point-of-care detection of poisons and has great potential as a tool in field applications.

Evaluation of Water Ammonium Ion Test Kit and Its Feasibility for the Analysis of Protein in Food.

The traditional method for the determination of protein in food needs the operations of digestion, distillation, absorption, and titration; therefore, it is complicated and time-consuming and requires professional personnel. Is there a more convenient and faster detection method that can directly determine the ammonium ions in protein digestion solution to obtain the protein content of food and avoid the distillation–absorption–titration process? The feasibility of water ammonium ion test kits for food protein rapid detection was discussed here. After digestion, the protein in food transforms into ammonium ions in the digestion solution. Because of the variety of food, there are many different inorganic ions left in the food digestion solution, and at the same time, digestion agents are added in the digestion process and become potential interference factors in ammonium determination. Therefore, the detection accuracy of ammonium test kits needs to be evaluated first, including their anti-interference ability. The standard curve of ammonium was established by the test kit. When the ammonium concentration was 0.00–2.50 mg/L, the absorbance at 620 nm was linearly related to the ammonium concentration, the determination coefficient R2 was 0.9995, and the detection limit of this method was 0.01 mg/L. The influences of temperature, pH value, and reaction time on the test kit method were discussed. The precision was 0.90–3.33%; the repeatability was 1.71–4.86%; and the recovery rate of tap water, river water, and sea water was controlled within 90–103%. The anti-interference ability of the evaluated test kit was better than that of the national standard detection method. The test kit, combined with sample pretreatment and protein conversion formula, was used to detect protein in different types of food (milk powder, rice flour, wheat flour, soy, banana, milk, fish food, chicken food, and dog food). The results showed that there were no significant differences (ρ > 0.05) between the national method and the test kit method. The ammonium ion test kit method shortened the determination time and had higher sensitivity, showing its potential for the rapid determination of food protein.

A colorimetric electronic tongue for point-of-care detection of COVID-19 using salivary metabolites

The monitoring of profile concentrations of chemical markers in saliva samples can be used to diagnose COVID-19 patients, and differentiate them from healthy individuals. Here, this purpose is achieved by designing a paper-based colorimetric sensor with an origami structure, containing general receptors such as pH-sensitive organic dyes, Lewis donors or acceptors, functionalized nanoparticles, and ion metal complexes. The color changes taking place in the receptors in the presence of chemical markers are visually observed and recorded with a digital instrument. Different types and amounts of the chemical markers provide the sensor with a unique response for patients (60 samples) or healthy (55 samples) individuals. These two categories can be discriminated with 84.3% accuracy. This study evidences that the saliva composition of cured and healthy participants is different from each other with accuracy of 85.7%. Moreover, viral load values obtained from the rRT-PCR method can be estimated by the designed sensor. Besides COVID-19, it may possible to simultaneously identify smokers and people with kidney disease and diabetes using the specified electronic tongue. Due to its high efficiency, the prepared paper device can be employed as a rapid detection kit to detect COVID-19.

Development of highly specific peptibody against Porcine epidemic diarrhea virus

Abstract Porcine epidemic diarrhea virus (PEDV) is an etiological agent of porcine epidemic diarrhea (PED), causing dehydration, vomiting, and acute watery diarrhea. Because it has a high mortality rate in neonatal piglets, PEDV is one of the critical pathogens affecting the swine industry. Since PEDV has shown rapid spreading in herds of swine, early diagnosis of this virus is very important to control the disease. Serological diagnosis test is a simple, rapid method for detecting PEDV infected pigs, and the development of PEDV-specific antibodies is essential for an accurate serological diagnostic test. Because conventional methods of development of monoclonal antibodies are time and hand-consuming work, recombinant antibodies are considered as an alternative strategy for the development of target-specific monoclonal antibodies. Peptibody is one of the promising recombinant antibodies that target-specific peptide is fused with the Fc domain of immunoglobulin G. The peptibody which have a specificity of peptides and immune function of Fc region as advantages have developed for therapeutic and diagnostic purpose. Here, we generated a PEDV-specific peptibody (PEDV-PB) that is fused with PEDV-specific peptide and mouse IgG2b Fc region. PEDV-specific peptide sequences were discovered by random phage display library and PEDV-PB was produced from Chinese hamster ovary (CHO) cells. PEDV-PB showed highly specific binding in several serological tests for PEDV, such as western blotting, immunofluorescence assay, and enzyme-linked immunosorbent assay. Our results suggest that PEDV-PB is very useful for the development of a rapid detection kit for PEDV.

Development of an Indirect ELISA Kit for Rapid Detection of Varicella-Zoster Virus Antibody by Glycoprotein E.

Varicella-zoster virus (VZV), a highly infectious agent that causes varicella (chickenpox), can also cause zoster (shingles), a disorder that is frequently associated with severe neuralgia. A reliable serological VZV diagnostic assay would be useful for identifying unprotected individuals and for surveilling post-vaccination immunoprotection status. Toward this goal, VZV membrane glycoprotein E (gE), the immunodominant VZV protein, served as target antigen in an indirect ELISA kit developed here to detect anti-VZV antibodies in clinical samples. For target antigen preparation, Chinese hamster ovary (CHO) cells were modified to express and secrete the VZV gE ectodomain, which was subsequently purified and used as coating antigen in an indirect ELISA. Ultimately, the optimal purified gE coating antigen concentration was determined to be 2 μg.ml−1 and the OD450nm detection cutoff value was 0.286. The coefficient of variation (CV) of intra-assay and inter-assay were <10 and 15%, respectively. A comparative test of 66 clinical samples showed that the coincidence rate was 93.9% between the indirect ELISA and a commercial varicella-zoster virus IgG ELISA kit. Thus, the indirect ELISA kit developed here may be useful for achieving rapid, sensitive, and specific detection of anti-VZV antibodies.

The Empowerment of Livestock Farmers in Subclinical Mastitis Test with GAMA Anti-Haptoglobin in “Sahabat Ternak” Etawah Crossbreed Goat Farm

Sahabat Ternak is one of the Etawah crossbreed (PE) goat farm groups in Sleman. This farm group focuses on goat milk production and processed goods. Problems that commonly arise in dairy goat farms are the cases of subclinical mastitis, which are quite high. This disease may cause a decrease in milk production and quality. The mastitis subclinical detection method which is often used by the farming community is the somatic cell count (SCC) and California mastitis test (CMT). However, both tests have low accuracy. Recently, a new method named GAMA Anti-Haptoglobin, which is more accurate and can be done easily has been established by livestock farmers. This community service aims to empower livestock farmers in applying GAMA Anti-Haptoglobin as a sensitive, rapid, and accurate subclinical mastitis detection kit in Sahabat Ternak goat farm. The method used in this activity consisted of discussion, socialization, and training for livestock farmers, as well as laboratory testing, evaluation of test findings, and treatment for PE goats. After the training, the livestock farmers were able to apply GAMA Anti-Haptoglobin mastitis detection method effectively. The implementation of this easy and accurate field mastitis detection method, as well as personnel with reliable skills, will support in the decrease of mastitis cases and increase milk production and quality, as well as community welfare.

Preparation of polyclonal antibodies against chemically synthesized ApxIA and ApxIVA toxins and their diagnostic efficacy in the experimentally injected mice

ObjectivesApxIA and ApxIVA are the main virulence factors of Actinobacillus pleuropneumoniae (A. pleuropneumoniae). The preparation of their antibodies is of great significance for diagnosing and treating porcine pleuropneumonia. MethodsThis study used chemically synthesized ApxIA and ApxIVA peptides as immunogens to prepare polyclonal antibodies against ApxIA and ApxIVA. Their titers were determined by ELISA and Dot blotting. Histopathology of apparently infected organs was carried out. Finally, the immunofluorescence method detected the distribution of antigens in the visceral tissues of mice infected with ApxIA and ApxIVA toxins. ResultsThe results showed that ApxIA and ApxIVA polyclonal antibodies with titer as high as 1: 200000 and 1: 50000, respectively, were successfully obtained. The immunofluorescence results showed that these polyclonal antibodies could accurately detect ApxIA and ApxIVA antigens in mouse lungs with strong specificity. Histopathologically, there were perialveolar hemorrhages, pulmonary emphysema, and sloughing of bronchiolar epithelium. There was vacuolation and infiltration of chronic inflammatory cells in the liver parenchymal tissue. Atrophy of intestinal villi and glands and sloughing of villus epithelium, and congestion. There was increased urinary space and congestion in the proximal convoluted tubules. Lymphoid follicles increased in size in the spleen, the germinal center enlarged, and congestion in surrounding lymphoid tissue were observed. ConclusionThis study laid a foundation for the research and development of a rapid detection kit for A. pleuropneumoniae and provided conditions for diagnosing and treating porcine contagious pleuropneumonia.

Dengue Immunological Markers Evolution at Saint Camille Hospital in Ouagadougou (HOSCO) Burkina Faso

&lt;b&gt;Background and Objective:&lt;/b&gt; Dengue is a remerging vector-borne viral disease in Burkina Faso since the outbreak of 2013 and requires special attention from health authorities. This study reports the prevalence of dengue fever serological markers (NS1Ag, IgM and IgG) and infection dynamic from January, 2018 to December, 2020 among patients tested for dengue infection at Saint Camille Hospital of Ouagadougou (HOSCO). &lt;b&gt;Materials and Methods:&lt;/b&gt; The study population consisted of 6414 patients aged 0-97 years. Dengue virus infection was detected in serum or plasma using the SD bioline dengue duo rapid detection kit. &lt;b&gt;Results:&lt;/b&gt; The prevalence of dengue NS1Ag was 2.25% (45/2003), 18.43% (501/2719) and 2.42% (38/1569) in the study population in 2018, 2019 and 2020, respectively. The age groups over 50 years and 15-20 years were significantly more infected compared to the group 21-30 years respectively in 2019 (p = 0.030) and 2020 (p = 0.035). Patients tested positive for at least one of these markers (NSlAg, IgG and IgM) represented 26.01% (521/2003) and 38.98% (1060/2719). The peak of infection during 2018 and 2019 was observed between October and November. The present study reports a high seroprevalence of acute dengue virus infection. The presence of NS1Ag, IgM and IgG in patients suggests an active circulation of the dengue virus in Ouagadougou. &lt;b&gt;Conclusion:&lt;/b&gt; Data shows recurrent outbreaks of dengue infection in our country need strong surveillance and a suitable and affordable diagnostic system to clarify the burden, pinpoint the risk factors and for better case management.

Health risk assessment of antimicrobial residues in sheep carcasses marketed in Kuwait

A total of 450 samples comprising 150 each of muscles, livers and kidneys were collected from 150 sheep carcasses in Kuwait and tested by both Rapid Premi®Test kits for rapid detection of antimicrobial-positive samples, and the High-performance liquid chromatography (HPLC) for residual determination of amoxicillin, tetracycline, oxytetracycline, and tylosin. Premi test revealed that 82%, 64% and 100% of the muscle, liver and kidney samples examined, respectively were positive for antimicrobials. HPLC analysis revealed mean concentrations (µg/kg) of 45.26, 148.17, 103.18, and 71.80 for amoxicillin, oxytetracycline, tetracycline, and tylosin respectively in muscles; 64.43, 263.15, 177.04, and 112.94, respectively in livers; and 53.12, 368.21, 196.40, and 138.63, respectively in kidneys. Although many samples exceeded the maximum residue limit (MRL) of Codex Alimentarius, the assessment of dietary exposure to the antimicrobials tested through consumption of sheep meat and organs did not constitute any health hazards in the different age groups of Kuwaiti population.