高级检索
当前位置: 首页 > 详情页

Multiplex competitive annealing mediated isothermal amplification with high fidelity DNA polymerase (HiFi-CAMP)

文献详情

资源类型:
Pubmed体系:
机构: [1]Shanghai Public Health Clinical Center, Fudan University, Shanghai, 201508, China [2]Medical Laboratory of Taizhou Fourth People’s Hospital, Taizhou, 225300, China [3]Department of Dermatology and Venereology, First Affiliated Hospital of Kunming Medical University, Kunming, 650032, China
出处:
ISSN:

关键词: HiFi-CAMP Isothermal amplification HFman probe Multiplex detection High-fidelity DNA polymerase Non-specific amplification

摘要:
Various isothermal amplification methods have been developed for point-of-care testing (POCT) of various infectious diseases. Here, we proposed a novel isothermal amplification method, named as 5'-half complementary primers mediated isothermal amplification (HCPA). Because of the similarity of our method to the previous method competitive annealing mediated isothermal amplification (CAMP) in primer design, we also use the name CAMP for our method. We demonstrated that CAMP is mediated by both a linear isothermal amplification pattern and a loop-mediated isothermal amplification pattern. To improve the specificity and enable multiplex detection, we further developed HiFi-CAMP method that uses a small amount of high-fidelity DNA polymerase to cut HFman probe to release fluorescent signal. The HiFi-CAMP method was demonstrated to have a good specificity and sensitivity, and fast amplification speed in detection of three human respiratory viruses, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), respiratory syncytial virus A (RSV-A) and influenza A viruses (IAV). When compared with gold standard RT-qPCR assays, the HiFi-CAMP assays showed sensitivities of 90.0 %, 71.4 % and 78.1 %, specificities of 100 %, 100 % and 95.5 %, and consistencies of 93.0 %, 93.3 % and 88.2 % for SARS-CoV-2, RSV-A and IAV, respectively. Furthermore, a duplex HiFi-CAMP assay was also developed to simultaneously detect RSV-A and SARS-CoV-2. The HiFi-CAMP will provide a promising candidate for POCT diagnosis in resource-limited settings.Copyright © 2024 Elsevier B.V. All rights reserved.

基金:
语种:
PubmedID:
中科院(CAS)分区:
出版当年[2025]版:
最新[2023]版:
大类 | 1 区 化学
小类 | 1 区 分析化学
第一作者:
第一作者机构: [1]Shanghai Public Health Clinical Center, Fudan University, Shanghai, 201508, China
通讯作者:
推荐引用方式(GB/T 7714):
APA:
MLA:

资源点击量:52537 今日访问量:0 总访问量:1562 更新日期:2024-09-01 建议使用谷歌、火狐浏览器 常见问题

版权所有©2020 昆明医科大学第一附属医院 技术支持:重庆聚合科技有限公司 地址:云南省昆明市西昌路295号(650032)