Next Article in Journal
Sensitivity Equalization and Dynamic Range Expansion with Multiple Optofluidic Microbubble Resonator Sensors
Previous Article in Journal
Advanced Textile-Based Wearable Biosensors for Healthcare Monitoring
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Biosensors
Volume 13
Issue 10
10.3390/bios13100910
biosensors-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

utPCR: A Strategy for the Highly Specific and Absolutely Quantitative Detection of Single Molecules within Only Minutes

by
Rui Wang
1,2,
Ying Liu
1,
Shuaiwei Chen
1,
Linlin Bai
1,
Kaiming Guo
1,
Yanan Pang
3,
Feng Qian
1,
Yongfang Li
4,
Li Ding
3,* and
Yongming Wang
1,5,*
1
State Key Laboratory of Genetic Engineering, School of Life Sciences, Zhongshan Hospital, Human Phenome Institute, Pudong Hospital, Fudan University, Shanghai 200438, China
2
School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA
3
Changhai Hospital, Second Military Medical University, Shanghai 200433, China
4
School of Food Science and Engineering, Foshan University, Foshan 528231, China
5
Shanghai Engineering Research Center of Industrial Microorganisms, Shanghai 200438, China
*
Authors to whom correspondence should be addressed.
Biosensors 2023, 13(10), 910; https://doi.org/10.3390/bios13100910
Submission received: 3 September 2023 / Revised: 23 September 2023 / Accepted: 25 September 2023 / Published: 27 September 2023
(This article belongs to the Section Biosensors and Healthcare)
Browse Figures
Versions Notes

Abstract

Bloodstream infection is a major health problem worldwide, with extremely high mortality. Detecting infection in the early stage is challenging due to the extremely low concentration of bacteria in the blood. Digital PCR provides unparalleled sensitivity and can achieve absolute quantification, but it is time-consuming. Moreover, the presence of unavoidable background signals in negative controls poses a significant challenge for single-molecule detection. Here, we propose a novel strategy called “Ultrafast flexible thin tube-based droplet digital PCR (utPCR)” that can shorten the digital PCR process from 2 h to only 5 min, with primer annealing/extension time reduced from minutes to only 5 s. Importantly, the ultrafast PCR eliminates nonspecific amplification and thus enables single-molecule detection. The utPCR enabled the sensitive detection and digital quantification of E. coli O157 in the high background of a 106-fold excess of E. coli K12 cells. Moreover, this method also displayed the potential to detect rare pathogens in blood samples, and the limit of detection (LOD) could be as low as 10 CFU per mL of blood without false positive results. Considered ultrafast (<5 min) and highly sensitive (single-molecule detection), the utPCR holds excellent prospects in the next generation of molecular diagnosis.
Keywords: ultrafast PCR; droplet digital PCR (ddPCR); single-molecule detection; bloodstream infection; absolute quantification; molecular diagnosis ultrafast PCR; droplet digital PCR (ddPCR); single-molecule detection; bloodstream infection; absolute quantification; molecular diagnosis

Share and Cite

MDPI and ACS Style

Wang, R.; Liu, Y.; Chen, S.; Bai, L.; Guo, K.; Pang, Y.; Qian, F.; Li, Y.; Ding, L.; Wang, Y. utPCR: A Strategy for the Highly Specific and Absolutely Quantitative Detection of Single Molecules within Only Minutes. Biosensors 2023, 13, 910. https://doi.org/10.3390/bios13100910

AMA Style

Wang R, Liu Y, Chen S, Bai L, Guo K, Pang Y, Qian F, Li Y, Ding L, Wang Y. utPCR: A Strategy for the Highly Specific and Absolutely Quantitative Detection of Single Molecules within Only Minutes. Biosensors. 2023; 13(10):910. https://doi.org/10.3390/bios13100910

Chicago/Turabian Style

Wang, Rui, Ying Liu, Shuaiwei Chen, Linlin Bai, Kaiming Guo, Yanan Pang, Feng Qian, Yongfang Li, Li Ding, and Yongming Wang. 2023. "utPCR: A Strategy for the Highly Specific and Absolutely Quantitative Detection of Single Molecules within Only Minutes" Biosensors 13, no. 10: 910. https://doi.org/10.3390/bios13100910

APA Style

Wang, R., Liu, Y., Chen, S., Bai, L., Guo, K., Pang, Y., Qian, F., Li, Y., Ding, L., & Wang, Y. (2023). utPCR: A Strategy for the Highly Specific and Absolutely Quantitative Detection of Single Molecules within Only Minutes. Biosensors, 13(10), 910. https://doi.org/10.3390/bios13100910

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop