Abstract
Highly sensitive (high SBR) and highly specific (high SNP discrimination ratio) DNA hybridization is essential for a biosensor with clinical application. Herein, we propose a method that allows detecting multiple pathogens on a single platform with the SNP discrimination ratios over 160:1 in the dynamic range of 101 to 104 copies per test. The newly developed SWAT method allows achieving highly sensitive and highly specific DNA hybridizations. The detection and discrimination of the MTB and NTM strain in the clinical samples with the SBR and SNP discrimination ratios higher than 160:1 indicate the high clinical applicability of the SWAT.
Highlights
Biosensors based on concurrent DNA-DNA hybridization for the simultaneous detection of various bacterial strains of the same species have unparallel importance in selecting accurate drug therapy [1,2]
The sensitivity of the DNA biosensors depends on two factors: (i) the signal to background ratio (SBR) and (ii) the specific to non-specific hybridization ratio, which is called the single nucleotide polymorphism (SNP) discrimination ratio
In conclusion, the highly sensitive and specific hybridization of the target DNA of diagnostic importance with probes is an essential criterion for the clinical applicability of diagnostic methods based on DNA hybridizations
Summary
Biosensors based on concurrent DNA-DNA hybridization for the simultaneous detection of various bacterial strains of the same species have unparallel importance in selecting accurate drug therapy [1,2]. For any DNA biosensor to be highly accurate and specific in detecting and discriminating against bacterial genotypes, the SBR and SNP discrimination ratio should be sufficiently high [5,6]. Achieving a significantly higher SNP discrimination ratio for direct clinical applications is challenging [12] Several approaches, such as the longer hybridization time (in hours), use of an elevated hybridization [13,14,15] and washing temperatures [16,17,18,19] close to the temperature of melting (Tm, ◦C), and high concentrations of chaotropic agents in hybridization buffers, have been used to increase the specificity [20,21]. High temperature decreases SBR due to the lower hybridization yield of the target DNA with the immobilized probe
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