TY - JOUR
T1 - Highly Specific and Rapid Multiplex Identification of Candida Species Using Digital Microfluidics Integrated with a Semi-Nested Genoarray
AU - Mao, Zeyin
AU - Deng, Anni
AU - Jin, Xiangyu
AU - Zhou, Tianqi
AU - Zhang, Shuailong
AU - Li, Meng
AU - Lv, Wenqi
AU - Huang, Leyang
AU - Zhong, Hao
AU - Wang, Shihong
AU - Shi, Yixuan
AU - Zhang, Lei
AU - Liao, Qinping
AU - Fu, Rongxin
AU - Huang, Guoliang
N1 - Publisher Copyright:
© 2024 American Chemical Society.
PY - 2024/11/26
Y1 - 2024/11/26
N2 - Candida species are the most common cause of fungal infections around the world, associated with superficial and even deep-seated infections. In clinical practice, there is great significance in identifying different Candida species because of their respective characteristics. However, current technologies have difficulty in onsite species identification due to long turnover time, high cost of reagents and instruments, or limited detection performance. We developed a semi-nested recombinase polymerase amplification (RPA) genoarray as well as an integrated system for highly specific identification of four Candida species with a simple design of primers, high detection sensitivity, fast turnover time, and good cost-effectiveness. The system constructed to perform the assay consists of a rapid sample processing module for nucleic acid release from fungal samples in 15 min and a digital microfluidic platform for precise and efficient detection reactions in 35 min. Therefore, our system could automatically identify specific Candida species, with a reagent consumption of only 2.5 μL of the RPA reaction mixture per target and no cross-reaction. Its detection sensitivity for four Candida species achieved 101-102 CFU/mL, which was 10-fold better than conventional RPA and even comparable to a common polymerase chain reaction. Evaluated by using cultured samples and 24 clinical samples, our system shows great applicability to onsite multiplex nucleic acid analysis.
AB - Candida species are the most common cause of fungal infections around the world, associated with superficial and even deep-seated infections. In clinical practice, there is great significance in identifying different Candida species because of their respective characteristics. However, current technologies have difficulty in onsite species identification due to long turnover time, high cost of reagents and instruments, or limited detection performance. We developed a semi-nested recombinase polymerase amplification (RPA) genoarray as well as an integrated system for highly specific identification of four Candida species with a simple design of primers, high detection sensitivity, fast turnover time, and good cost-effectiveness. The system constructed to perform the assay consists of a rapid sample processing module for nucleic acid release from fungal samples in 15 min and a digital microfluidic platform for precise and efficient detection reactions in 35 min. Therefore, our system could automatically identify specific Candida species, with a reagent consumption of only 2.5 μL of the RPA reaction mixture per target and no cross-reaction. Its detection sensitivity for four Candida species achieved 101-102 CFU/mL, which was 10-fold better than conventional RPA and even comparable to a common polymerase chain reaction. Evaluated by using cultured samples and 24 clinical samples, our system shows great applicability to onsite multiplex nucleic acid analysis.
UR - http://www.scopus.com/inward/record.url?scp=85209258126&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.4c04265
DO - 10.1021/acs.analchem.4c04265
M3 - Article
AN - SCOPUS:85209258126
SN - 0003-2700
VL - 96
SP - 18797
EP - 18805
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 47
ER -