Your browser doesn't support javascript.
loading
The End of the Gray Zone: One-Tube Nested Recombinase Polymerase Amplification with Ultrahigh Signal-to-Noise Ratio for Precisely Detecting and Surveilling Viruses.
Cao, Gaihua; Xiong, Yifan; Shi, Meimei; Qiu, Yue; Bian, Yong; Nie, Fuping; Huo, Danqun; Hou, Changjun.
Afiliación
  • Cao G; Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, Sichuan 400044, PR China.
  • Xiong Y; State Key Laboratory of Cattle Diseases Detection (Chongqing) of Customs. Diagnosis and Testing Laboratory of Lumpy Skin Disease, Chongqing Customs Technology Center, Chongqing 400020, PR China.
  • Shi M; Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, Sichuan 400044, PR China.
  • Qiu Y; State Key Laboratory of Cattle Diseases Detection (Chongqing) of Customs. Diagnosis and Testing Laboratory of Lumpy Skin Disease, Chongqing Customs Technology Center, Chongqing 400020, PR China.
  • Bian Y; State Key Laboratory of Cattle Diseases Detection (Chongqing) of Customs. Diagnosis and Testing Laboratory of Lumpy Skin Disease, Chongqing Customs Technology Center, Chongqing 400020, PR China.
  • Nie F; Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, Sichuan 400044, PR China.
  • Huo D; State Key Laboratory of Cattle Diseases Detection (Chongqing) of Customs. Diagnosis and Testing Laboratory of Lumpy Skin Disease, Chongqing Customs Technology Center, Chongqing 400020, PR China.
  • Hou C; Science and Technology Research Center of China Customs, Beijing 100730, PR China.
Anal Chem ; 95(27): 10414-10421, 2023 07 11.
Article en En | MEDLINE | ID: mdl-37367936
The samples were difficult to accurately determine positive or negative between 35 and 40 cycles by real-time quantitative PCR (qPCR) as the standard method. Here, we developed one-tube nested recombinase polymerase amplification (ONRPA) technology with CRISPR/Cas12a to overcome this difficulty. ONRPA broke the amplification plateau to substantially enhance the signals, which considerably improved the sensitivity and eliminated the problem of gray area. Using two pairs of primers one after another, it improved precision by lowering the probability of magnifying several target zones, which was completely free of contamination by nonspecific amplification. This was important in nucleic acid testing. Finally, by the CRISPR/Cas12a system as a terminal output, the approach achieved a high signal output as few as 2.169 copies·µL-1 in 32 min. ONRPA was 100-fold more sensitive than conventional RPA and 1000-fold compared to qPCR. ONRPA coupled with CRISPR/Cas12a will be an important and new promoter of RPA in clinical applications.
Asunto(s)
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Amplificación de Ácido Nucleico / Recombinasas Tipo de estudio: Diagnostic_studies Idioma: En Revista: Anal Chem Año: 2023 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas de Amplificación de Ácido Nucleico / Recombinasas Tipo de estudio: Diagnostic_studies Idioma: En Revista: Anal Chem Año: 2023 Tipo del documento: Article Pais de publicación: Estados Unidos