Fast and visual detection of nucleic acids using a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM
Anal Chim Acta. 2023 Apr 1;1248:340938. doi: 10.1016/j.aca.2023.340938. Epub 2023 Feb 3.ABSTRACTCRISPR-Cas12a (Cpf1) is widely used for pathogen detection. However, most Cas12a nucleic acid detection methods are limited by a PAM sequence requirement. Moreover, preamplification and Cas12a cleavage are separate. Here, we developed a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM sequence with high sensitivity and specificity that offers one-tube, rapid, and visually observable detection of nucleic acids. In this system, Cas12a detection and RPA amplification are performed simultaneously, without separate...
Source: Analytica Chimica Acta - February 22, 2023 Category: Chemistry Authors: Kangfeng Lin Jianguang Guo Xiangju Guo Qinghan Li Xiao Li Zhen Sun Zehang Zhao Jiao Weng Jinzhun Wu Rui Zhang Boan Li Source Type: research
Fast and visual detection of nucleic acids using a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM
Anal Chim Acta. 2023 Apr 1;1248:340938. doi: 10.1016/j.aca.2023.340938. Epub 2023 Feb 3.ABSTRACTCRISPR-Cas12a (Cpf1) is widely used for pathogen detection. However, most Cas12a nucleic acid detection methods are limited by a PAM sequence requirement. Moreover, preamplification and Cas12a cleavage are separate. Here, we developed a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM sequence with high sensitivity and specificity that offers one-tube, rapid, and visually observable detection of nucleic acids. In this system, Cas12a detection and RPA amplification are performed simultaneously, without separate...
Source: Analytica Chimica Acta - February 22, 2023 Category: Chemistry Authors: Kangfeng Lin Jianguang Guo Xiangju Guo Qinghan Li Xiao Li Zhen Sun Zehang Zhao Jiao Weng Jinzhun Wu Rui Zhang Boan Li Source Type: research
Fast and visual detection of nucleic acids using a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM
Anal Chim Acta. 2023 Apr 1;1248:340938. doi: 10.1016/j.aca.2023.340938. Epub 2023 Feb 3.ABSTRACTCRISPR-Cas12a (Cpf1) is widely used for pathogen detection. However, most Cas12a nucleic acid detection methods are limited by a PAM sequence requirement. Moreover, preamplification and Cas12a cleavage are separate. Here, we developed a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM sequence with high sensitivity and specificity that offers one-tube, rapid, and visually observable detection of nucleic acids. In this system, Cas12a detection and RPA amplification are performed simultaneously, without separate...
Source: Analytica Chimica Acta - February 22, 2023 Category: Chemistry Authors: Kangfeng Lin Jianguang Guo Xiangju Guo Qinghan Li Xiao Li Zhen Sun Zehang Zhao Jiao Weng Jinzhun Wu Rui Zhang Boan Li Source Type: research
Fast and visual detection of nucleic acids using a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM
Anal Chim Acta. 2023 Apr 1;1248:340938. doi: 10.1016/j.aca.2023.340938. Epub 2023 Feb 3.ABSTRACTCRISPR-Cas12a (Cpf1) is widely used for pathogen detection. However, most Cas12a nucleic acid detection methods are limited by a PAM sequence requirement. Moreover, preamplification and Cas12a cleavage are separate. Here, we developed a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM sequence with high sensitivity and specificity that offers one-tube, rapid, and visually observable detection of nucleic acids. In this system, Cas12a detection and RPA amplification are performed simultaneously, without separate...
Source: Analytica Chimica Acta - February 22, 2023 Category: Chemistry Authors: Kangfeng Lin Jianguang Guo Xiangju Guo Qinghan Li Xiao Li Zhen Sun Zehang Zhao Jiao Weng Jinzhun Wu Rui Zhang Boan Li Source Type: research
Fast and visual detection of nucleic acids using a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM
Anal Chim Acta. 2023 Apr 1;1248:340938. doi: 10.1016/j.aca.2023.340938. Epub 2023 Feb 3.ABSTRACTCRISPR-Cas12a (Cpf1) is widely used for pathogen detection. However, most Cas12a nucleic acid detection methods are limited by a PAM sequence requirement. Moreover, preamplification and Cas12a cleavage are separate. Here, we developed a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM sequence with high sensitivity and specificity that offers one-tube, rapid, and visually observable detection of nucleic acids. In this system, Cas12a detection and RPA amplification are performed simultaneously, without separate...
Source: Analytica Chimica Acta - February 22, 2023 Category: Chemistry Authors: Kangfeng Lin Jianguang Guo Xiangju Guo Qinghan Li Xiao Li Zhen Sun Zehang Zhao Jiao Weng Jinzhun Wu Rui Zhang Boan Li Source Type: research
Fast and visual detection of nucleic acids using a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM
Anal Chim Acta. 2023 Apr 1;1248:340938. doi: 10.1016/j.aca.2023.340938. Epub 2023 Feb 3.ABSTRACTCRISPR-Cas12a (Cpf1) is widely used for pathogen detection. However, most Cas12a nucleic acid detection methods are limited by a PAM sequence requirement. Moreover, preamplification and Cas12a cleavage are separate. Here, we developed a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM sequence with high sensitivity and specificity that offers one-tube, rapid, and visually observable detection of nucleic acids. In this system, Cas12a detection and RPA amplification are performed simultaneously, without separate...
Source: Analytica Chimica Acta - February 22, 2023 Category: Chemistry Authors: Kangfeng Lin Jianguang Guo Xiangju Guo Qinghan Li Xiao Li Zhen Sun Zehang Zhao Jiao Weng Jinzhun Wu Rui Zhang Boan Li Source Type: research
Fast and visual detection of nucleic acids using a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM
Anal Chim Acta. 2023 Apr 1;1248:340938. doi: 10.1016/j.aca.2023.340938. Epub 2023 Feb 3.ABSTRACTCRISPR-Cas12a (Cpf1) is widely used for pathogen detection. However, most Cas12a nucleic acid detection methods are limited by a PAM sequence requirement. Moreover, preamplification and Cas12a cleavage are separate. Here, we developed a one-step RPA-CRISPR detection (ORCD) system unrestricted by the PAM sequence with high sensitivity and specificity that offers one-tube, rapid, and visually observable detection of nucleic acids. In this system, Cas12a detection and RPA amplification are performed simultaneously, without separate...
Source: Analytica Chimica Acta - February 22, 2023 Category: Chemistry Authors: Kangfeng Lin Jianguang Guo Xiangju Guo Qinghan Li Xiao Li Zhen Sun Zehang Zhao Jiao Weng Jinzhun Wu Rui Zhang Boan Li Source Type: research
Generating enhanced mucosal immunity against Bordetella pertussis: current challenges and new directions
Bordetella pertussis (Bp) is the highly transmissible etiologic agent of pertussis, a severe respiratory disease that causes particularly high morbidity and mortality in infants and young children. Commonly known as “whooping cough,” pertussis is one of the least controlled vaccine-preventable diseases worldwide with several countries experiencing recent periods of resurgence despite broad immunization coverage. While current acellular vaccines prevent severe disease in most cases, the immunity they confer wanes rapidly and does not prevent sub clinical infection or transmission of the bacterium to new and vulnerable h...
Source: Frontiers in Immunology - February 21, 2023 Category: Allergy & Immunology Source Type: research
Modeling the Effect of Oxidative Stress on Bordetella pertussis Fermentations
AbstractA mathematical model is proposed forBordetella pertussis with the main goal to better understand and describe the relation between cell growth, oxidative stress and NADPH levels under different oxidative conditions. The model is validated with flask experiments conducted under different conditions of oxidative stress induced by high initial glutamate concentrations, low initial inoculum and secondary culturing following exposure to starvation. The model exhibited good accuracy when calibrated and validated for the different experimental conditions. From comparisons of model predictions to data with different model ...
Source: Biotechnology Progress - February 17, 2023 Category: Biotechnology Authors: Michael Vitelli,
I. Melih Tamer,
Mark Pritzker,
Hector Budman Tags: RESEARCH ARTICLE Source Type: research
Deep longitudinal multi-omics analysis of Bordetella pertussis cultivated in bioreactors highlights medium starvations and transitory metabolisms, associated to vaccine antigen biosynthesis variations and global virulence regulation
Bordetella pertussis is the bacterial causative agent of whooping cough, a serious respiratory illness. An extensive knowledge on its virulence regulation and metabolism is a key factor to ensure pertussis vaccine manufacturing process robustness. The aim of this study was to refine our comprehension of B. pertussis physiology during in vitro cultures in bioreactors. A longitudinal multi-omics analysis was carried out over 26 h small-scale cultures of B. pertussis. Cultures were performed in batch mode and under culture conditions intending to mimic industrial processes. Putative cysteine and proline starvations were, resp...
Source: Frontiers in Microbiology - February 14, 2023 Category: Microbiology Source Type: research
Novel murine model reveals an early role for pertussis toxin in disrupting neonatal immunity to Bordetella pertussis
The increased susceptibility of neonates to specific pathogens has previously been attributed to an underdeveloped immune system. More recent data suggest neonates have effective protection against most pathogens but are particularly susceptible to those that target immune functions specific to neonates. Bordetella pertussis (Bp), the causative agent of “whooping cough”, causes more serious disease in infants attributed to its production of pertussis toxin (PTx), although the neonate-specific immune functions it targets remain unknown. Problematically, the rapid development of adult immunity in mice has confounded our ...
Source: Frontiers in Immunology - February 8, 2023 Category: Allergy & Immunology Source Type: research
A semi high-throughput whole blood-based flow cytometry assay to detect and monitor Bordetella pertussis-specific Th1, Th2 and Th17 responses
DiscussionThus, this assay appears as a promising test for evaluation of Bp antigen-specific CD4+ T cells induced by current and next generation pertussis vaccines. (Source: Frontiers in Immunology)
Source: Frontiers in Immunology - February 6, 2023 Category: Allergy & Immunology Source Type: research
Hand disinfectants and their activity against clinical isolates of Bordetella pertussis
CONCLUSION: Products A and B were assessed as effective against clinical isolates of B. pertussis in accordance with EN 1500. Quaternary ammonium compound-based product C did not comply with the requirements of EN 1500.PMID:36718925 | DOI:10.21101/cejph.a7141 (Source: Central European Journal of Public Health)
Source: Central European Journal of Public Health - January 31, 2023 Category: International Medicine & Public Health Authors: Petra Uttlov á Jan Urban Source Type: research
