The construction of Mycobacterium tuberculosis 16S rDNA MSPQC sensor based on Exonuclease III-assisted cyclic signal amplification

Add time:09/03/2019    Source:sciencedirect.com

Tuberculosis caused by Mycobacterium tuberculosis (M. tuberculosis) remains one of the most serious infectious diseases all over the world. The key to reduce the spread and mortality rate of tuberculosis is to develop faster and more sensitive approaches for detection of M. tuberculosis. However, current detection methods can not meet the requirements of rapid clinical M. tuberculosis detection in terms of detection time. Herein, a new 16S rDNA multichannel series piezoelectric quartz crystal (MSPQC) sensor based on Exonuclease III (Exo III)-aided target recycling has been developed for rapid detection of M. tuberculosis. The specific 16S rDNA fragment of M. tuberculosis was used as biomarker, DNA capture probes complementary to the biomarker were designed and modified on the surface of AuNPs. The Exo III which could recognise hybrid duplexes and selectively digest DNA capture probe was used to assist digestion cycle by digesting DNA capture probe and releasing the intact target fragment. After all DNA probes loading on the surface of AuNPs were removed, the surface of AuNPs was exposed and conductive connection was formed between the nanogap network electrode by self-catalytic growth of exposed AuNPs in the glucose and HAuCl4 solution. This resulted in sensitive response of M. tuberculosis sensor and M. tuberculosis was detected by recording this response. The limit of detection (LOD) of the method was 20 CFU/mL and the detection time was less than 3 h. It was expected to be widely used in detection methods of M. tuberculosis.

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