A Commercial Glucose Meter for Portable in vitro Molecular Diagnosis of Hepatitis B Virus

doi:10.19894/j.issn.1000-0518.210182

Chinese Journal of Applied Chemistry ›› 2022, Vol. 39 ›› Issue (3): 498-506.DOI: 10.19894/j.issn.1000-0518.210182

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A Commercial Glucose Meter for Portable in vitro Molecular Diagnosis of Hepatitis B Virus

Jia-Xue YU¹^,², Chang WANG²^,³, Mei-Ting YANG², Yan DU²^,³(), Chang LIU¹()

^1.College of Pharmacy，Jinzhou Medical University，Jinzhou 121001，China
^2.State Key Laboratory of Electroanalytical Chemistry，Changchun Institute of Applied Chemistry，Chinese Academy of Sciences，Changchun 130022，China
^3.School of Applied Chemistry and Engineering，University of Science & Technology of China，Hefei 230026，China

Received:2020-04-12 Accepted:2020-07-30 Published:2022-03-01 Online:2022-03-15
Contact: Yan DU,Chang LIU
About author:liuchang@jzmu.edu.cn；duyan@ciac.ac.cn
Supported by:
the National Natural Science Foundation of China(21874129);the International Technological Cooperation Project of Jilin Scientific and Technological Development Program(20200801044GH)

Abstract

Abstract:

The current detection methods for hepatitis B virus （HBV） require high cost， complicated manipulation and are usually with high misdiagnosis rate. All the above issues have inspired us to develop a portable biosensing platform for the HBV detection. The loop-mediated isothermal amplification （LAMP） reaction designed for HBV sequence contributes to the single copy gene detection within 40 min. The immobilization of capture probe F-Probe （FP） on the magnetic beads （MBs） and the modification of FPc （partially complementary to FP） with heat-resistant invertase （Inv） collectively constitute the Inv-FPc signal probe， and the hybridization of Inv-FPc and FP creates the signal transduction probe， which with the HBV LAMP amplicons would induce the replacement of nucleic acid chains， isolating the Inv-FPc from the signal transduction probe. The Inv-FPc signal probe in the solution after magnetic separation would catalyze the transformation of sucrose into glucose that is read by glucometer. The experimental results show that the qualified signal response is in the concentration range of 10~500 nmol/L of HBV gene segments. Almost concordant signal and background are observed when employing Norovirus （NoV） gene as the target. The biosensor presents a perfect “yes/no” signal response in 10% human serum and has a potential to be applied in point of care diagnosis of HBV.

Key words: Loop-mediated isothermal amplification, Glucometer, Hepatitis B virus, Point-of-care testing

CLC Number:

O655.9

Jia-Xue YU, Chang WANG, Mei-Ting YANG, Yan DU, Chang LIU. A Commercial Glucose Meter for Portable in vitro Molecular Diagnosis of Hepatitis B Virus[J]. Chinese Journal of Applied Chemistry, 2022, 39(3): 498-506.

Figures/Tables 6

Table 1 Oligonucleotide sequences used in this work

名称	DNA序列（5' to 3'）
Name	DNA sequences （5' to 3'）
HBV?BIP	TTGGATGTTGGGGTACTTTACCGAGGTCTATTTACAGGCAGTT
HBV?FIP	TCCCATGAAGTTAAGGGAGTAGCAATTTTCTTTTGTCTTTGGGTA
HBV?F3	TTGAGTCCCTTTTTACCTCTA
HBV?B3	TCTCTGACATACTTTCCAATCA
HBV?LP	CAAGAACATATTGTACTAAAAATCAAGCAATGTTTTCGAA
HBV mimic	TACATTTAAACCCTAATAAAACCAAACGTTGGGG
NoV?BIP	ATCAATCACTGAAGGGCTCCCCTGAGGAGCCAATGAGCAATG
NoV?FIP	TCCATCACGCTGGGAGATAGGAATTTTCCCCAGAACCACACC
NoV?B3	ACCTCAGAGAGTGCACAGAG
NoV?F3	CTGCAGCCCTGGAGATCA
NoV mimic	TCTGGGGTGCCTTGCACTTCACAATGGAATTC
FPc	CGACCCCAACGTTTGGTTTTATTAGGGTTTAAATGTAAAAAAAAAAA/SH?SH C6/
FP	TAATAAAACCAAACGTTGGGGTCGAAAAAAAAAA/biotin/

Fig.1 Schematic illustration of coupling loop mediated isothermal amplification with nucleic acid circuit to glucometer for HBV detection

Fig.2 （A） The kinetics of enzyme reaction at different temperatures；（B） Signal-to-noise ratio at different reaction temperature

Fig.3 （A） Signal response of glucometer to different concentrations of HBV mimic from 0~500 nmol/L；（B） Selectivity and sensitivity detection of different concentration of HBV and NoV mimics

Fig.4 （A） The function of time with varying amounts of the HBV templates；（B） Agarose gel electrophoretic characterization of LAMP amplicons

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A Commercial Glucose Meter for Portable in vitro Molecular Diagnosis of Hepatitis B Virus

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