TY - JOUR
T1 - Polymerase chain reaction-based ultrasensitive detection of HBV DNA via G-quadruplex selective iridium(III) complex luminescent probe
AU - Li, Zongbing
AU - Zou, Seyin
AU - Wu, Shujie
AU - Miao, Xiangmin
AU - Ma, Edmond Dik Lung
N1 - Funding Information:
This work was supported by the Natural Science Foundation of Xuzhou City (KC18140), the Traditional Chinese Medicine Bureau of Guangdong Province (20202004), Guangdong Second Provincial General Hospital 2017 Youth Fund Project (TQ2017-008), Hong Kong Baptist University (FRG2/17?18/003), the Health and Medical Research Fund (HMRF/14150561), the National Natural Science Foundation of China (21575121 and 21775131), the Hong Kong Baptist University Century Club Sponsorship Scheme 2019, the Interdisciplinary Research Matching Scheme (RC?IRMS/16?17/03), Interdisciplinary Research Clusters Matching Scheme (RC?IRCs/17?18/03), Collaborative Research Fund (C5026?16G), SKLEBA and HKBU Strategic Development Fund (SKLP_1920_P02).
Funding Information:
This work was supported by the Natural Science Foundation of Xuzhou City ( KC18140 ), the Traditional Chinese Medicine Bureau of Guangdong Province ( 20202004 ), Guangdong Second Provincial General Hospital 2017 Youth Fund Project (TQ2017-008), Hong Kong Baptist University ( FRG2/17–18/003 ), the Health and Medical Research Fund (HMRF/14150561), the National Natural Science Foundation of China ( 21575121 and 21775131 ), the Hong Kong Baptist University Century Club Sponsorship Scheme 2019, the Interdisciplinary Research Matching Scheme (RC–IRMS/16–17/03), Interdisciplinary Research Clusters Matching Scheme (RC–IRCs/17–18/03), Collaborative Research Fund (C5026–16G), SKLEBA and HKBU Strategic Development Fund ( SKLP_1920_P02 ).
Copyright © 2020 Elsevier B.V. All rights reserved.
PY - 2021/1/1
Y1 - 2021/1/1
N2 - Polymerase chain reaction (PCR) is the gold standard for low-abundant DNA detection. Here, to expand the application of PCR with novel detecting methods, we developed a label-free fluorescent sensor for ultrasensitive and one-step detection of hepatitis B virus (HBV) DNA using the G-quadruplex selective iridium(III) complex luminescent probe. By using HBV DNA as the template with two hairpin structure primers that contained oxyethylene glycol tethers, PCR amplification occurred and generated numbers of specific PCR products with free G-quadruplex sequences at both ends. Such free G-quadruplex sequences can change into G-quadruplex structure with the help of K+, resulting in a strong luminescence intensity upon their binding with the G-quadruplex selective iridium(III) complex. The luminescence intensity increase was proportional to the concentration of PCR products, and indirectly related with HBV DNA concentration. Moreover, the utilization of the iridium(III) complex effectively improved the specificity of the sensor, while PCR paved the way for the ultrasensitive detection of DNA in the linear range of 3.0 fM to 800 pM, with a detection limit of 1.6 fM. Notably, this assay was successfully used to detect HBV DNA in normal and patient serum samples, indicating a potential application for biomolecular analysis.
AB - Polymerase chain reaction (PCR) is the gold standard for low-abundant DNA detection. Here, to expand the application of PCR with novel detecting methods, we developed a label-free fluorescent sensor for ultrasensitive and one-step detection of hepatitis B virus (HBV) DNA using the G-quadruplex selective iridium(III) complex luminescent probe. By using HBV DNA as the template with two hairpin structure primers that contained oxyethylene glycol tethers, PCR amplification occurred and generated numbers of specific PCR products with free G-quadruplex sequences at both ends. Such free G-quadruplex sequences can change into G-quadruplex structure with the help of K+, resulting in a strong luminescence intensity upon their binding with the G-quadruplex selective iridium(III) complex. The luminescence intensity increase was proportional to the concentration of PCR products, and indirectly related with HBV DNA concentration. Moreover, the utilization of the iridium(III) complex effectively improved the specificity of the sensor, while PCR paved the way for the ultrasensitive detection of DNA in the linear range of 3.0 fM to 800 pM, with a detection limit of 1.6 fM. Notably, this assay was successfully used to detect HBV DNA in normal and patient serum samples, indicating a potential application for biomolecular analysis.
KW - Fluorescent sensor
KW - G-quadruplex
KW - Hepatitis B virus DNA
KW - Iridium(III) complex
KW - Polymerase chain reaction
UR - http://www.scopus.com/inward/record.url?scp=85091331385&partnerID=8YFLogxK
U2 - 10.1016/j.talanta.2020.121661
DO - 10.1016/j.talanta.2020.121661
M3 - Journal article
C2 - 33076171
AN - SCOPUS:85091331385
SN - 0039-9140
VL - 221
JO - Talanta
JF - Talanta
M1 - 121661
ER -