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1.
Anal Chem ; 96(15): 5752-5756, 2024 Apr 16.
Article in English | MEDLINE | ID: mdl-38560822

ABSTRACT

Viruses are the primary cause of many infectious diseases in both humans and animals. Various testing methods require an amplification step of the viral RNA sample before detection, with quantitative reverse transcription polymerase chain reaction (RT-qPCR) being one of the most widely used along with lesser-known methods like Nucleic Acid Sequence-Based Amplification (NASBA). NASBA offers several advantages, such as isothermal amplification and high selectivity for specific sequences, making it an attractive option for low-income facilities. In this research, we employed a single electrochemical biosensor (E-Biosensor) designed for potentially detecting any virus by modifying the NASBA protocol. In this modified protocol, a reverse primer is designed with an additional 22-nucleotide sequence (tag region) at the 5'-end, which is added to the NASBA process. This tag region becomes part of the final amplicon generated by NASBA. It can hybridize with a single specific E-Biosensor probe set, enabling subsequent virus detection. Using this approach, we successfully detected three different viruses with a single E-Biosensor design, demonstrating the platform's potential for virus detection.


Subject(s)
Biosensing Techniques , Viruses , Animals , Humans , Sensitivity and Specificity , Self-Sustained Sequence Replication/methods , RNA, Viral/genetics , RNA, Viral/analysis , Viruses/genetics , Nucleic Acid Amplification Techniques
2.
Anal Methods ; 16(5): 772-779, 2024 02 01.
Article in English | MEDLINE | ID: mdl-38230437

ABSTRACT

A highly reproducible electrochemical biosensor, employing a five-stranded four-way junction (5S-4WJ) system through square wave voltammetry, has been successfully validated for the detection of Influenza A virus (InfA). A comprehensive assessment of its linearity, precision, accuracy, and robustness has demonstrated its compliance with FDA standards. Integration with Nucleic Acid-Based Amplification (NASBA) has showcased its selectivity for InfA, enabling the detection of InfA RNA with a standard heater set at 41 °C. This platform offers a straightforward setup well-suited for use at low-resource facilities.


Subject(s)
Biosensing Techniques , Influenza A virus , Influenza A virus/genetics , RNA , Nucleic Acid Amplification Techniques
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