Article
Food Science & Technology
Lin Ding, Xiaoli Xu, Xiaofu Wang, Xiaoyun Chen, Yuwen Lu, Junfeng Xu, Cheng Peng
Summary: Effective regulation and resolution of public concerns are prerequisites for the industrialization of gene-edited crops and derived foods. This study successfully established qualitative and quantitative PCR assays for detecting Cpf1, a commonly used target site in gene editing. The methods were found to be specific, sensitive, and stable, providing technical support for the future monitoring of gene-edited products and their derived foods.
Article
Immunology
Xiaolin Jiang, Yong Jiang, Feng Ye
Summary: This study used real-time quantitative PCR to detect Mucorales and Aspergillus in formalin-fixed, paraffin-embedded samples. The combination of Mucorales RQ-PCR and Aspergillus RQ-PCR was recommended as a screening tool for detecting suspected mucormycosis and/or aspergillosis samples.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2023)
Article
Chemistry, Analytical
Andrei Hutanu, Chiara Signori, Bernd Moritz, Manuel Gregoritza, Adelheid Rohde, Maria A. Schwarz
Summary: In this study, a capillary electrophoresis-based gel free hybridization assay using fluorescently labeled peptide nucleic acids (PNAs) is proposed to evaluate the identity of nucleic acid species in gene therapy products. Various proof-of-concept studies demonstrate the potential of PNA probes for advanced analytical characterization of therapeutic modalities like oligonucleotides, plasmids, mRNA, and DNA released by recombinant adeno-associated virus. The method is highly specific for single-stranded nucleic acids up to 1000 nucleotides, with a limit of quantification in the picomolar range when multiple probes are used.
ANALYTICAL CHEMISTRY
(2023)
Article
Biochemistry & Molecular Biology
Kathannan Sankar, Hyung Joo Yoon, Young Bo Lee, Kyeong Yong Lee
Summary: Bumble bees are important alternative pollinators and model insects. This study identified stable reference genes for the bumble bee species and their different lines and tissues using RT-qPCR procedures and four analysis programs. The results emphasized the need to evaluate the stability of candidate reference genes for any differently designed lines and tissue conditions in bumble bee species.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Genetics & Heredity
Cristina Martinez-Fernandez de la Camara, Michelle E. McClements, Robert E. MacLaren
Summary: This study found that the conformation of the DNA standard significantly affects the accuracy of absolute quantification by qPCR, with the use of supercoiled undigested plasmid DNA template generating a higher titer. Contrary to previous studies, pre-treating samples with Proteinase K after DNase I digestion and using a high temperature step resulted in a reduction in AAV titers.
Review
Obstetrics & Gynecology
Tomohiro Mitoma, Kei Hayata, Satomi Yokohata, Akiko Ohira, Chiaki Kashino, Satoe Kirino, Kazumasa Tani, Jota Maki, Eriko Eto, Hisashi Masuyama
Summary: Cases of uterine wall thinning and placental abnormalities complicated with systemic lupus erythematosus during pregnancy have been reported in Asian countries for ten years. Clinicians should pay attention to the possibility of uterine wall thinning as it may lead to uterine atony and structural abnormality of the placenta in SLE patients.
BMC PREGNANCY AND CHILDBIRTH
(2022)
Article
Biochemistry & Molecular Biology
Zhiwei Chen, Nigel G. Halford, Chenghong Liu
Summary: Real-time quantitative PCR is a technique for measuring the content of a specific nucleic acid sequence in a sample, and it can be divided into absolute and relative quantitative methods. Relative quantification is mainly used for gene expression in functional genomics and transcriptome studies. However, to accurately use this technology, key points such as designing specific primers, selecting appropriate reference genes, and performing scientific calculations and statistics are crucial. This article proposes a workflow for relative quantitative PCR and introduces relevant points to help beginners better understand and utilize this technology.
Article
Horticulture
Chenjie Zhang, Chengwei Song, Linfeng Chen, Huili Ma, Yabing Zhang, Dalong Guo, Lili Guo, Xiaogai Hou
Summary: The aim of this study was to identify suitable reference genes for normalizing the expression levels of miRNA in different developmental stages and tissues. Five algorithms were used for stability analysis, and the results identified mtr-MIR160b-p3 and gma-miR394a-5p as the most stable miRNAs during bud development in early-flowering tree peony 'Feng dan', while PC-5p-19095 was the most stable during bud development in late-flowering tree peony 'Lian he'. Additionally, mtr-miR159a was found to be the most stable miRNA during flower development stages of different tree peony varieties.
Article
Infectious Diseases
Tae Kwon Kim, Jessica Waldman, Freddy Ibanez-Carrasco, Lucas Tirloni, Camila Waltero, Christiano Calixo, Gloria R. Braz, Albert Mulenga, Itabajara da Silva Vaz, Carlos Logullo
Summary: This study identified and selected stable reference genes in the R. microplus genome and evaluated their expression stability during embryogenesis. Results showed that Rpl4, Elf1a, and GAPDH are suitable internal controls for normalizing qPCR during R. microplus embryogenesis.
TICKS AND TICK-BORNE DISEASES
(2023)
Article
Cell Biology
Gloria B. Kim, Jens Fritsche, Sebastian Bunk, Andrea Mahr, Felix Unverdorben, Kevin Tosh, Hong Kong, Colby R. Maldini, Chui Lau, Sriram Srivatsa, Shuguang Jiang, Joshua Glover, Derek Dopkin, Carolyn X. Zhang, Heiko Schuster, Daniel J. Kowalewski, Valentina Goldfinger, Martina Ott, David Fuhrmann, Maike Baues, Hans Boesmueller, Christoph Schraeder, Gisela Schimmack, Colette Song, Franziska Hoffgaard, Michael Roemer, Chih-Chiang Tsou, Martin Hofmann, Thomas Treiber, Meike Hutt, Leonie Alten, Maike Jaworski, Amir Alpert, Sarah Missel, Carsten Reinhardt, Harpreet Singh, Oliver Schoor, Steffen Walter, Claudia Wagner, Dominik Maurer, Toni Weinschenk, James L. Riley
Summary: By analyzing a large number of tumor and normal tissue samples using immunopeptidomics, a highly expressed tumor antigen in multiple solid cancers was identified. Further research discovered that by enhancing the affinity, this specific TCR can be transduced into T cells and eliminate tumors in vivo.
SCIENCE TRANSLATIONAL MEDICINE
(2022)
Article
Microbiology
Shuaijie Li, Kui Guo, Xuefeng Wang, Yuezhi Lin, Jinhui Wang, Yaoxin Wang, Cheng Du, Zhe Hu, Xiaojun Wang
Summary: A tat-gag-based real-time quantitative PCR (TG-qPCR) was developed as an effective tool for early diagnosis of clinical Equine Infectious Anemia (EIA) disease. The TG-qPCR showed high specificity, sensitivity, and reproducibility, and had better inclusivity compared to the traditional qPCR method.
MICROBIOLOGY SPECTRUM
(2023)
Article
Food Science & Technology
Ling Li, Yuanmiao Wei, Yao Liu, Shuna Xiang, Hanyue Zhang, Ying Shang
Summary: In this study, the matB gene was identified as a species-specific gene of L. edodes, serving as an ideal reference gene for qualitative and quantitative PCR assays to detect L. edodes material. This provides a convenient and accurate method for detecting L. edodes products and adulteration in wild edible mushroom products.
FOOD SCIENCE & NUTRITION
(2022)
Article
Food Science & Technology
Yao Liu, Ling Li, Yuanmiao Wei, Hanyue Zhang, Shuna Xiang, Ying Shang
Summary: The article introduces a detection method for T. albuminosus mushrooms using the TSA gene as an endogenous reference gene. The method is validated through qualitative and quantitative PCR, providing a fast and accurate way to detect T. albuminosus and protect consumers' rights while promoting the industry.
LWT-FOOD SCIENCE AND TECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Yao Dai, Yumeng Cao, Jens Kohler, Aiping Lu, Shaohua Xu, Haiyun Wang
Summary: This study identified OAZ1 and hsa-miR-6835-3p as the most reliable individual reference genes for mRNA and miRNA quantification in cancer exosomes. A combination of reference genes - OAZ1/SERF2/MPP1 for mRNA and hsa-miR-6835-3p/hsa-miR-4468-3p for miRNA analyses is recommended for superior accuracy.
Article
Chemistry, Analytical
Reona Takabatake, Tomohiro Egi, Keisuke Soga, Jumpei Narushima, Satoko Yoshiba, Norihito Shibata, Kosuke Nakamura, Kazunari Kondo, Masahiro Kishine, Junichi Mano, Kazumi Kitta
Summary: Many countries have implemented labeling systems for genetically modified organisms (GMOs). Japan is revising and restricting the regulatory threshold for non-GMO labeling. A qualitative method based on comparative Cq-based analysis has been developed in this study, which could be useful for the revised non-GMO labeling.
ANALYTICAL CHEMISTRY
(2022)
