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1دورية أكاديمية
المؤلفون: Emese Pekker, Katalin Priskin, Éva Szabó-Kriston, Bernadett Csányi, Orsolya Buzás-Bereczki, Lili Adorján, Valéria Szukacsov, Lajos Pintér, Miklós Rusvai, Paul Cooper, Endre Kiss-Tóth, Lajos Haracska
المصدر: Biological Procedures Online, Vol 25, Iss 1, Pp 1-14 (2023)
مصطلحات موضوعية: Canine, Mesenchymal stem cells, Cell therapy, Pathogen testing, EMA guidelines, Validation, Medicine (General), R5-920, Biology (General), QH301-705.5
الوصف: Abstract Background The action of mesenchymal stem cells (MSCs) is the subject of intense research in the field of regenerative medicine, including their potential use in companion animals, such as dogs. To ensure the safety of canine MSC batches for their application in regenerative medicine, a quality control test must be conducted in accordance with Good Manufacturing Practices (GMP). Based on guidance provided by the European Medicines Agency, this study aimed to develop and validate a highly sensitive and robust, nucleic acid-based test panel for the detection of various canine pathogens. Analytical sensitivity, specificity, amplification efficiency, and linearity were evaluated to ensure robust assessment. Additionally, viable spike-in controls were used to control for optimal nucleic acid extraction. The conventional PCR-based and real-time PCR-based pathogen assays were evaluated in a real-life setting, by direct testing MSC batches. Results The established nucleic acid-based assays displayed remarkable sensitivity, detecting 100–1 copies/reaction of template DNA. They also exhibited high specificity and efficiency. Moreover, highly effective nucleic acid isolation was confirmed by the sensitive detection of spike-in controls. The detection capacity of our optimized and validated methods was determined by direct pathogen testing of nine MSC batches that displayed unusual phenotypes, such as reduced cell division or other deviating characteristics. Among these MCS batches of uncertain purity, only one tested negative for all pathogens. The direct testing of these samples yielded positive results for important canine pathogens, including tick-borne disease-associated species and viral members of the canine infectious respiratory disease complex (CIRDC). Notably, samples positive for the etiological agents responsible for enteritis (CPV), leptospirosis (Leptospira interrogans), and neosporosis (Neospora caninum) were also identified. Furthermore, we conducted biosafety evaluation of 12 MSC batches intended for therapeutic application. Eleven MSC batches were found to be free of extraneous agents, and only one tested positive for a specific pathogen, namely, canine parvovirus. Conclusion In this study, we established and validated reliable, highly sensitive, and accurate nucleic acid-based testing methods for a broad spectrum of canine pathogens.
وصف الملف: electronic resource
العلاقة: https://doaj.org/toc/1480-9222Test
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2دورية أكاديمية
المؤلفون: Shuaiwei Chen, Cui Wu, Chunyan Qian, Yanan Pang, Kaiming Guo, Ting Wang, Linlin Bai, Feng Qian, Zunzhong Ye, Zhenping Liu, Zhaohui Qiao, Yongming Wang, Rui Wang
مصطلحات موضوعية: Biophysics, Biochemistry, Medicine, Cell Biology, Molecular Biology, Biotechnology, Immunology, Computational Biology, Space Science, Biological Sciences not elsewhere classified, Mathematical Sciences not elsewhere classified, Information Systems not elsewhere classified, red ” strategy, high background interference, automatically analyzed using, care pathogen testing, care diagnosis versatile, based method coupled, positive detection rate, entire detection process, portable cartridge capable, based “ green, 2 variants using, method yielded results, care detection, “ green, portable cartridge, method achieved, simultaneous detection, multiplex detection
الوصف: Versatile, informative, sensitive, and specific nucleic acid detection plays a crucial role in point-of-care pathogen testing, genotyping, and disease monitoring. In this study, we present a novel one-pot Cas12b-based method coupled with the “Green-Yellow-Red” strategy for multiplex detection. By integrating RT-LAMP amplification and Cas12b cleavage in a single tube, the entire detection process can be completed within 1 h. Our proposed method exhibits high specificity, enabling the discrimination of single-base mutations with detection sensitivity approaching single molecule levels. Additionally, the fluorescent results can be directly observed by the naked eye or automatically analyzed using our custom-designed software Result Analyzer. To realize point-of-care detection, we developed a portable cartridge capable of both heating and fluorescence excitation. In a clinical evaluation involving 20 potentially SARS-CoV-2-infected samples, our method achieved a 100% positive detection rate when compared to standard RT-PCR. Furthermore, the identification of SARS-CoV-2 variants using our method yielded results that were consistent with the sequencing results. Notably, our proposed method demonstrates excellent transferability, allowing for the simultaneous detection of various pathogens and the identification of mutations as low as 0.5% amidst a high background interference. These findings highlight the tremendous potential of our developed method for molecular diagnostics.
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3دورية أكاديمية
المؤلفون: Manjunath Keremane, Khushwant Singh, Chandrika Ramadugu, Robert R. Krueger, Todd H. Skaggs
المصدر: Plants, Vol 13, Iss 3, p 411 (2024)
مصطلحات موضوعية: citrus germplasm, budwood certification, Next-Generation Sequencing, pathogen testing, biological indexing, real time PCR, Botany, QK1-989
الوصف: Citrus is affected by many diseases, and hence, the movement of citrus propagative materials is highly regulated in the USA. Currently used regulatory pathogen detection methods include biological and laboratory-based technologies, which are time-consuming, expensive, and have many limitations. There is an urgent need to develop alternate, rapid, economical, and reliable testing methods for safe germplasm exchange. Citrus huanglongbing (HLB) has devastated citrus industries leading to an increased need for germplasm exchanges between citrus growing regions for evaluating many potentially valuable hybrids for both HLB resistance and multilocational performance. In the present study, Next-Generation Sequencing (NGS) methods were used to sequence the transcriptomes of 21 test samples, including 15 well-characterized pathogen-positive plants. A workflow was designed in the CLC Genomics Workbench software, v 21.0.5 for bioinformatics analysis of the sequence data for the detection of pathogens. NGS was rapid and found to be a valuable technique for the detection of viral and bacterial pathogens, and for the discovery of new citrus viruses, complementary to the existing array of biological and laboratory assays. Using NGS methods, we detected beet western yellows virus, a newly reported citrus virus, and a variant of the citrus yellow vein-associated virus associated with the “fatal yellows” disease.
وصف الملف: electronic resource
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4دورية أكاديمية
المؤلفون: Stephen M. Rich, Eric L. Siegel, Guang Xu
المصدر: Journal of Clinical Medicine, Vol 12, Iss 20, p 6522 (2023)
مصطلحات موضوعية: human-biting tick, pathogen testing, tick-borne disease, Medicine
الوصف: With expanding concern about ticks, there is a general sense of uncertainty about the diagnosis and treatment of tick-borne diseases. The diagnosis process is often based on clinical judgment in conjunction with laboratory testing and can be pathogen specific. Treatments may require disease-dependent approaches, and co-infections complicate or increase the severity of the clinical picture. Measuring exposure indices in the tick has become popular among providers and their patients, though this practice is not universally understood, and certain public health agencies have voiced concerns regarding interpretation and rigor of testing. As many providers subscribe to or recommend these services to aid in pretest risk and exposure assessments, this work sought to clarify the role of pathogen testing human-biting ticks as a complement to the diagnostic pipeline and raises points that must be addressed through future research and interdisciplinary conversation. Future work is needed to develop quality control oversight for tick testing laboratories. Studies on the integration of tick testing with human cases to see how these services affect health outcomes are also needed. Alongside these, improvements in the quality and availability of diagnostics are of critical importance.
وصف الملف: electronic resource
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5دورية أكاديمية
المؤلفون: Rich, Stephen M., Siegel, Eric L., Xu, Guang
المصدر: Microbiology Department Faculty Publication Series
مصطلحات موضوعية: human-biting tick, pathogen testing, tick-borne disease
الوصف: With expanding concern about ticks, there is a general sense of uncertainty about the diagnosis and treatment of tick-borne diseases. The diagnosis process is often based on clinical judgment in conjunction with laboratory testing and can be pathogen specific. Treatments may require disease-dependent approaches, and co-infections complicate or increase the severity of the clinical picture. Measuring exposure indices in the tick has become popular among providers and their patients, though this practice is not universally understood, and certain public health agencies have voiced concerns regarding interpretation and rigor of testing. As many providers subscribe to or recommend these services to aid in pretest risk and exposure assessments, this work sought to clarify the role of pathogen testing human-biting ticks as a complement to the diagnostic pipeline and raises points that must be addressed through future research and interdisciplinary conversation. Future work is needed to develop quality control oversight for tick testing laboratories. Studies on the integration of tick testing with human cases to see how these services affect health outcomes are also needed. Alongside these, improvements in the quality and availability of diagnostics are of critical importance.
وصف الملف: application/pdf
العلاقة: https://scholarworks.umass.edu/micro_faculty_pubs/351Test; https://scholarworks.umass.edu/context/micro_faculty_pubs/article/1350/viewcontent/jcm_12_06522_v2.pdfTest
الإتاحة: https://doi.org/10.3390/jcm12206522Test
https://scholarworks.umass.edu/micro_faculty_pubs/351Test
https://scholarworks.umass.edu/context/micro_faculty_pubs/article/1350/viewcontent/jcm_12_06522_v2.pdfTest -
6دورية أكاديمية
المؤلفون: A. I. Barashkova, A. D. Reshetnikov, E. N. Popov
المصدر: Российский паразитологический журнал, Vol 15, Iss 3, Pp 17-22 (2021)
مصطلحات موضوعية: ixodes persulcatus, seasonal activity, tick collection, spermophilus parryii, central yakutia, pathogen testing, Biology (General), QH301-705.5
الوصف: The purpose of the research is to identify ixodid ticks and their pathogens in the left bank area of Central Yakutia. Materials and methods. The work was carried out in 2019–2020 in the left bank area of Central Yakutia. Nine ticks were collected in 2019, and 27 ticks in 2020. We studied forest shrub stations, steppe stations, meadow field stations, near-water stations and stations of settlements. To determine faunal and ecological characteristics of ectoparasites in the territory, we used standard collection methods. The tick species was determined using morphological keys by N. A. Filippova; the determination correctness was confirmed by the PCR method. The collected ticks were studied for causative agents of babesiosis and tick-borne viral encephalitis using PCR analysis. Results and discussion. One species of ixodid ticks, Ixodes persulcatus, inhabits the left bank area of Central Yakutia. Haemaphysalis concinna was not found in Yakutia. In 2008, a natural focus of blood protozoan disease of domestic reindeer appeared for the first time in Yakutia in its central zone. Recently, an increase in the number of I. persulcatus has been observed. Tick activity is recorded from the second decade of May to the second decade of August. The ground-squirrel Spermophilus parryii is the main host for the preimaginal stages. Pathogens were not detected when ticks were examined for causative agents of babesiosis and tick-borne viral encephalitis using PCR analysisd.
وصف الملف: electronic resource
العلاقة: https://vniigis.elpub.ru/jour/article/view/792Test; https://doaj.org/toc/1998-8435Test; https://doaj.org/toc/2541-7843Test
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7دورية أكاديمية
المؤلفون: Carelene Lakhan, Neela Badrie, Adash Ramsubhag, Lisa Indar
المصدر: Microorganisms; Volume 10; Issue 8; Pages: 1601
مصطلحات موضوعية: diarrhea, molecular diagnostic tool, enhanced foodborne pathogen testing, public health, Trinidad and Tobago
جغرافية الموضوع: agris
الوصف: In 2009, the burden of illness study for acute gastroenteritis in Trinidad and Tobago highlighted that ~10% of stool samples tested were positive for a foodborne pathogen. The study also noted that limited laboratory screening for pathogens contributed to a lack of etiology as public health hospitals only routinely tested for Salmonella and Shigella, and sometimes for Escherichia coli and Campylobacter. To better understand the foodborne pathogens responsible for acute gastroenteritis, enhanced testing using the BioFire® FilmArray® Gastrointestinal PCR panel was used to screen diarrheal stool samples for 22 pathogens from patients in 2018. The five general public health hospitals (San Fernando, Mt. Hope, Port of Spain, Sangre Grande, and Tobago) were notified of research activities and diarrheal stool samples were collected from all acute gastroenteritis patients. A total of 66 stools were screened and ~30% of samples tested positive for a foodborne pathogen. The current study showed that a much wider range of enteric pathogens were associated with acute gastroenteritis in Trinidad and Tobago than previously reported in 2009. These findings can be used by health officials to guide appropriate interventions, as well as to provide evidence for adoption of the PCR panel detection method at public health hospitals to benefit patient care.
وصف الملف: application/pdf
العلاقة: Food Microbiology; https://dx.doi.org/10.3390/microorganisms10081601Test
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8دورية أكاديمية
المؤلفون: Catherine D. Carrillo, Burton W. Blais
المصدر: Frontiers in Sustainable Food Systems, Vol 5 (2021)
مصطلحات موضوعية: whole-genome sequencing (WGS), Salmonella, validation, benchmark datasets, Listeria (L.) monocytogenes, food pathogen testing, Nutrition. Foods and food supply, TX341-641, Food processing and manufacture, TP368-456
الوصف: Whole-genome sequencing (WGS) technologies are rapidly being adopted for routine use in food microbiology laboratories worldwide. Examples of how WGS is used to support food safety testing include gene marker discovery (e.g., virulence and anti-microbial resistance gene determination) and high-resolution typing (e.g., cg/wgMLST analysis). This has led to the establishment of large WGS databases representing the genomes of thousands of different types of food pathogenic and commensal bacteria. This information constitutes an invaluable resource that can be leveraged to develop and validate routine test methods used to support regulatory and industry food safety objectives. For example, well-curated raw and assembled genomic datasets of the key food pathogens (Salmonella enterica, Listeria monocytogenes, and Shiga-toxigenic Escherichia coli) have been used in our laboratory in studies to validate bioinformatics pipelines, as well as new molecular methods as a prelude to the laboratory phase of the “wet lab” validation process. The application of genomic information to food microbiology method development will decrease the cost of test development and lead to the generation of more robust methodologies supporting risk assessment and risk management actions.
وصف الملف: electronic resource
العلاقة: https://www.frontiersin.org/articles/10.3389/fsufs.2021.754988/fullTest; https://doaj.org/toc/2571-581XTest
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9دورية أكاديمية
المؤلفون: Dawood Al-Ajmi, Shafeeq Rahman, Sharmila Banu
المصدر: BMC Microbiology, Vol 20, Iss 1, Pp 1-10 (2020)
مصطلحات موضوعية: Escherichia coli O157, Antimicrobial resistance, Food pathogen testing, Microbiology, QR1-502
الوصف: Abstract Background Shiga toxin-producing Escherichia coli (STEC) is a major source of food-borne illness around the world. E. coli O157 has been widely reported as the most common STEC serogroup and has emerged as an important enteric pathogen. Cattle, in particular have been identified as a major E. coli O157:H7 reservoir of human infections; however, the prevalence of this organism in camels, sheep, and goats is less understood. The aim of this study was to evaluate the occurrence and concentration of E. coli serotype O157 in the feces of healthy camels (n = 140), cattle (n = 137), sheep (n = 141) and goats (n = 150) slaughtered in United Arab Emirates (UAE) for meat consumption between September 2017 and August 2018. We used immunomagnetic separation coupled with a culture-plating method to detect E. coli O157. Non-sorbitol fermenting colonies were assessed via latex-agglutination testing, and positive cultures were analyzed by performing polymerase chain reactions to detect genes encoding attaching and effacing protein (eaeA), hemolysin A (hlyA, also known as ehxA) and Shiga toxin (stx1 and stx2), and E. coli O157:H7 specific genes (rfb O157, uidA, and fliC). All E. coli O157 isolates were analyzed for their susceptibility to 20 selected antimicrobials. Results E. coli O157 was observed in camels, goats, and cattle fecal samples at abundances of 4.3, 2, and 1.46%, respectively, but it was undetectable in sheep feces. The most prevalent E. coli O157 gene in all STEC isolates was stx 2 ; , whereas, stx 1 was not detected in any of the samples. The fecal samples from camels, goats, and cattle harbored E. coli O157 isolates that were 100% susceptible to cefotaxime, chloramphenicol, ciprofloxacin, norfloxacin, and polymyxin B. Conclusion To our knowledge, this is the first report on the occurrence of E. coli O157 in slaughter animals in the UAE. Our results clearly demonstrate the presence of E. coli O157 in slaughtered animals, which could possibly contaminate meat products intended for human consumption.
وصف الملف: electronic resource
العلاقة: http://link.springer.com/article/10.1186/s12866-020-01899-0Test; https://doaj.org/toc/1471-2180Test
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10دورية أكاديمية
المؤلفون: K. Sreenath, Priyam Batra, E. V. Vinayaraj, Ridhima Bhatia, KVP SaiKiran, Vishwajeet Singh, Sheetal Singh, Nishant Verma, Urvashi B. Singh, Anant Mohan, Sushma Bhatnagar, Anjan Trikha, Randeep Guleria, Rama Chaudhry
المصدر: Microbiology Spectrum, Vol 9, Iss 1 (2021)
مصطلحات موضوعية: COVID-19, SARS-CoV-2, coinfections, multiple-pathogen testing, respiratory PCR, Microbiology, QR1-502
الوصف: ABSTRACT Emerging evidence indicates that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected individuals are at an increased risk for coinfections; therefore, physicians need to be cognizant about excluding other treatable respiratory pathogens. Here, we report coinfection with SARS-CoV-2 and other respiratory pathogens in patients admitted to the coronavirus disease (COVID) care facilities of an Indian tertiary care hospital. From June 2020 through January 2021, we tested 191 patients with SARS-CoV-2 for 33 other respiratory pathogens using an fast track diagnostics respiratory pathogen 33 (FTD-33) assay. Additionally, information regarding other relevant respiratory pathogens was collected by reviewing their laboratory data. Overall, 13 pathogens were identified among patients infected with SARS-CoV-2, and 46.6% (89/191) of patients had coinfection with one or more additional pathogens. Bacterial coinfections (41.4% [79/191]) were frequent, with Staphylococcus aureus being the most common, followed by Klebsiella pneumoniae. Coinfections with SARS-CoV-2 and Pneumocystis jirovecii or Legionella pneumophila were also identified. The viral coinfection rate was 7.3%, with human adenovirus and human rhinovirus being the most common. Five patients in our cohort had positive cultures for Acinetobacter baumannii and K. pneumoniae, and two patients had active Mycobacterium tuberculosis infection. In total, 47.1% (90/191) of patients with coinfections were identified. The higher proportion of patients with coinfections in our cohort supports the systemic use of antibiotics in patients with severe SARS-CoV-2 pneumonia with rapid de-escalation based on respiratory PCR/culture results. The timely and simultaneous identification of coinfections can contribute to improved health of COVID-19 patients and enhanced antibiotic stewardship during the pandemic. IMPORTANCE Coinfections in COVID-19 patients may worsen disease outcomes and need further investigation. We found that a higher proportion of patients with COVID-19 were coinfected with one or more additional pathogens. A better understanding of the prevalence of coinfection with other respiratory pathogens in COVID-19 patients and the profile of pathogens can contribute to effective patient management and antibiotic stewardship during the current pandemic.
وصف الملف: electronic resource
العلاقة: https://doaj.org/toc/2165-0497Test
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