Article
Microbiology
Li Liu, Mengyao Li, Gaowen Liu, Jian He, Yang Liu, Xuesong Chen, Yungui Tu, Jie Lin, Yue Feng, Xueshan Xia
Summary: This study developed a highly sensitive and efficient method for the detection of brucellosis using a one-tube nested quantitative real-time PCR (qPCR) approach. The method showed an impressive analytical sensitivity of 100 fg/μL, surpassing conventional qPCR and allowing detection of low levels of Brucella DNA. An evaluation of 250 clinical samples demonstrated a specificity of 100% and a sensitivity of 98.6%, indicating its reliability and accuracy. Importantly, the new method significantly improved the detection rate of low-burden samples by reducing cycle threshold values by an average of 6.4. These findings highlight the immense potential of this approach for rapid and accurate brucellosis diagnosis, critical for disease management and control.
MICROBIOLOGY SPECTRUM
(2023)
Article
Chemistry, Analytical
Teruaki Tozaki, Aoi Ohnuma, Shinichi Iwai, Mio Kikuchi, Taichiro Ishige, Hironaga Kakoi, Keiichi Hirota, Kanichi Kusano, Shunichi Nagata
Summary: The study investigated the robustness of digital PCR and real-time PCR in transgene detection. Digital PCR was relatively robust when primer and probe sequences did not completely match the template transgene, while real-time PCR was less sensitive. Lowering the annealing temperature can improve PCR sensitivity.
ANALYTICAL CHEMISTRY
(2021)
Article
Developmental Biology
Yann W. Yap, Natalie J. Hannan, Euan M. Wallace, Sarah A. Marshall
Summary: Nrf proteins have been found to play a crucial role in placental-derived oxidative stress-induced pregnancy disorders. Successful silencing of Nrf proteins in human cells has been achieved for further research.
Article
Microbiology
Aline Lamien-Meda, Martina Koehsler, Julia Walochnik
Summary: This study aimed to establish a new qPCR assay for the detection and quantification of Acanthamoeba T4 genotype. The T4 assay showed good specificity and high detection efficiency, and it could be run simultaneously with the Qvarnstrom assay.
Article
Microbiology
Travis Richins, Katelyn Houghton, Joel Barratt, Sarah G. H. Sapp, Anna Peterson, Yvonne Qvarnstrom
Summary: Human-infecting Cyclospora spp. are coccidian parasites that cause cyclosporiasis, a gastrointestinal disease, in humans. The development of sensitive molecular tests has greatly improved the diagnosis of cyclosporiasis outbreaks and the understanding of its source. In this study, two novel one-tube nested qPCR assays were developed to detect human-infecting Cyclospora spp. in clinical stool samples. The assay targeting the cytb mitochondrial locus showed better performance compared to the routine 18S assay. This finding suggests that the cytb assay may be useful for diagnostic laboratories screening clinical fecal specimens suspected of containing human-infecting Cyclospora spp.
MICROBIOLOGY SPECTRUM
(2023)
Article
Biochemistry & Molecular Biology
Pablo Hernandez-Ortego, Remedios Torres-Montero, Elvira de la Pena, Felix Viana, Jorge Fernandez-Trillo
Summary: TRPM8 is a non-selective cation channel widely expressed in primary sensory neurons and other tissues. This study characterized the performance of six commercial TRPM8 antibodies in three immunodetection techniques and found that two antibodies were successful in detecting TRPM8 in all approaches, while the other four were only acceptable for specific techniques.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Cell Biology
Renjie Liao, Manas Mondal, Christopher D. Nazaroff, Diego Mastroeni, Paul D. Coleman, Joshua Labaer, Jia Guo
Summary: The method presented achieves highly sensitive and multiplexed protein analysis using cleavable fluorescent tyramide and off-the-shelf antibodies, enhancing detection sensitivity and reducing imaging time. By studying genetically identical cell populations and profiling neurons in human brain tissue, it reveals protein expression heterogeneity and the presence of distinct neuron clusters within different sub-regions of the hippocampus.
FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
(2021)
Article
Immunology
Prem Perumal, Mohamed Bilal Abdullatif, Harriet N. Garlant, Isobella Honeyborne, Marc Lipman, Timothy D. McHugh, Jo Southern, Ronan Breen, George Santis, Kalaiarasan Ellappan, Saka Vinod Kumar, Harish Belgode, Ibrahim Abubakar, Sanjeev Sinha, Seshadri S. Vasan, Noyal Joseph, Karen E. Kempsell
Summary: This study focused on validating mRNA biomarkers identified in a non-human primate TB model in new patient cohorts. The biomarkers showed good performance characteristics in diagnosing ATB and LTBI, and identifying high-risk LTBI individuals for progression, with potential for low-cost rapid diagnostic solutions.
FRONTIERS IN IMMUNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Kaiping Deng, Shizhen Steven Wang, Shannon Kiener, Emily Smith, Kai -Shun Chen, Ruiqing Pamboukian, Anna Laasri, Catalina Pelaez, Jodie Ulaszek, Matthew Kmet, Antonio De Jesus, Thomas Hammack, Ravinder Reddy, Hua Wang
Summary: The FDA has developed a rapid qPCR method for detecting Salmonella in food, which has been shown to be reproducible and sensitive.
Article
Agriculture, Dairy & Animal Science
Jinpeng Zhang, Huanru Fu, Cuiteng Chen, Jinxiu Jiang, Yusheng Lin, Bin Jiang, Lin Lin, Qilin Hu, Chunhe Wan
Summary: PiMeV infection was detected in fecal samples of pigeons and a sensitive and specific TaqMan-qPCR method was established. The study also suggests that PiMeV may be vertically transmitted in racing pigeon embryos.
Article
Neurosciences
Anne-Kathrin Lutz, Helen Friedericke Bauer, Valentin Ioannidis, Michael Schoen, Tobias M. Boeckers
Summary: This study validated a set of commercial and homemade SHANK3 antibodies in Western Blotting, synaptic immunocytochemistry, and immunohistochemistry, and found that only a small subset of the antibodies met the criteria of quality and specificity. The study highlights the importance of antibody specificity testing in the research field.
FRONTIERS IN SYNAPTIC NEUROSCIENCE
(2022)
Article
Biochemistry & Molecular Biology
Antonio Augusto Fonseca, Mateus Laguardia-Nascimento, Amanda Petrina Scota Ferreira, Carla do Amaral Pinto, Valdenia Lopes da Silva Goncalves, Aline Aparecida Silva Barbosa, Anselmo Vasconcelos Rivetti, Marcelo Fernandes Camargos
Summary: The objective of this study was to differentiate and correctly diagnose red sea bream iridovirus (RSIV) and infectious spleen and kidney necrosis virus (ISKNV) in fish farming. The sequencing of ORF450L, ORF342L, ORF077 and the intergenic region between ORF37 and ORF42R showed that ORF077 separated the three major clades of Megalocytivirus (MCV), while ISKNV could not be differentiated. All qPCRs tests showed acceptable repeatability values, less than 5%. Therefore, two ISKNV qPCRs and two RSIV qPCRs are considered suitable for the diagnosis and typing of MCV.
MOLECULAR BIOLOGY REPORTS
(2023)
Article
Biochemistry & Molecular Biology
Eva Kocsmar, Marlene Schmid, Miguel Cosenza-Contreras, Ildiko Kocsmar, Melanie Foell, Leah Krey, Balint Andras Barta, Gergely Racz, Andras Kiss, Martin Werner, Oliver Schilling, Gabor Lotz, Peter Bronsert
Summary: Protein expression is a main focus in histological diagnostics and tissue-based research. The post-mortem applicability of protein expression is still unclear. Autopsy tissue specimens provide unique insight into advanced disease states, especially in cancer research. In this study, the maximum post-mortem interval (PMI) suitable for characterizing protein expression patterns was identified. Significant protein degradation was observed in kidneys and livers at 48 hours, while degradation in lungs was detected at 72 hours, suggesting organ-specific degradation kinetics. Proteins with similar post-mortem kinetics are not primarily shared in their biological functions. Structural features may play a role in determining post-mortem stability.
CELLULAR AND MOLECULAR LIFE SCIENCES
(2023)
Article
Anatomy & Morphology
Teresa Lorenzi, Andrea Sagrati, Eva Montanari, Martina Senzacqua, Manrico Morroni, Mara Fabri
Summary: The study analyzed the distribution and morphology of nNOS positive neurons in the indusium griseum. Immunohistochemical and immunofluorescence techniques were used to process sagittal serial sections of corpus callosum from autopsy specimens, and the specificity of the antibody was tested. The presence and distribution of neuronal nitric oxyde synthase-immunopositive neurons suggest that indusium griseum likely plays a functional role in the neurovascular regulation within the corpus callosum.
BRAIN STRUCTURE & FUNCTION
(2022)
Article
Food Science & Technology
Sarah Azinheiro, Dipak Ghimire, Joana Carvalho, Marta Prado, Alejandro Garrido-Maestu
Summary: A multiplex RT-qPCR method was developed to specifically detect live bacteria, with two genetic targets co-amplified for improved specificity and an Internal Amplification Control to rule out false negative results due to reaction inhibition. The method demonstrated high sensitivity in smoked salmon samples inoculated with live and dead bacteria, showing almost complete concordance with the reference standard ISO 11290.