关键词: 关键词：DNA检测, 杂交捕获, 高通量, 核酸提取

Abstract: Objective To introduce a high throughput DNA detection technique based on capture, which not require nucleic acid extraction and suitable for various applications with different samples, to provide a powerful means for genetic analysis. Methods The whole blood, dry blood spots, saliva and oral swabs were lysed to release the DNA, after denaturation, DNA was captured to 96-well plates by hybridization. After washing off the unbound probe, DNA can be amplified by qPCR with specific primers, or enzymatic ligating the probes to form a single strand-template with specific sequence at both ends, and qPCR is carried out with universal primers. By using P16 gene as a target, the effect of this technique was evaluated, and the technique was applied to detect parasite DNA in whole blood, serum and dry blood spots, its sensitivity and specificity were evaluated. The stability of DNA detection was evaluated with saliva and oral swabs under different storage conditions. Results This method can be successfully applied to detect DNA in whole blood, dry blood spots, saliva, oral swabs and so on. The results of saliva and swabs are still stable after being stored at room temperature and 4 ℃ for 15 days. The limit of detection of this method for Plasmodium is 0.06 parasites / μL, which is more sensitive than traditional microscopic examination and can accurately detect the parasite DNA in infected samples. Conclusions A high-throughput DNA detection technique without nucleic acid extraction is developed, which is suitable for different sample types and targets. It can provide a sensitive and efficient tool for large scale gene screening analysis.

Key words: Key words: DNA detection, hybrid capture, high-throughput, nucleic acid extraction