RESUMEN
An integrated dual-signal bioassay was devised to fulfil thrombin (TB) ultrasensitive detection by integrating visualization with the photoelectrochemical technique based on G-quadruplex/hemin. During the process, branched sheet embedded copper-based oxides prepared with illumination and alkaline condition play a vital role in obtaining the desirable photocurrent. The switchover of photoelectrochemical signal was realized by the adjustable distance between electron acceptor G-quadruplex/hemin and interface materials due to dissociation of the Cu/Mn double-doped cerium dioxide (CuMn@CeO2)/DNA caused by the addition of TB. Then, CuMn@CeO2 transferred onto visual zones triggered catalytic reactions under the existence of 3,3',5,5'-tetramethylbenzidine and hydrogen peroxide, making a variation in color recognized by the naked eye and providing visual prediction. Under optimized conditions, this bioassay protocol demonstrated wide linear ranges (0.0001-50 nM), high selectivity, stability, and reproducibility. More importantly, the proposed visual/photoelectrochemical transduction mechanism platform exhibits a lower background signal and more reliable detection results, which also offers an effective way for detecting other proteins and nucleic acids.
Asunto(s)
Técnicas Biosensibles , Hemina , Reproducibilidad de los Resultados , Técnicas Biosensibles/métodos , Técnicas Electroquímicas/métodos , Límite de Detección , CobreRESUMEN
BACKGROUND: Anopheles sinensis is the major malaria vector in China and Southeast Asia. Vector control is one of the most effective measures to prevent malaria transmission. However, there is little transcriptome information available for the malaria vector. To better understand the biological basis of malaria transmission and to develop novel and effective means of vector control, there is a need to build a transcriptome dataset for functional genomics analysis by large-scale RNA sequencing (RNA-seq). METHODS: To provide a more comprehensive and complete transcriptome of An. sinensis, eggs, larvae, pupae, male adults and female adults RNA were pooled together for cDNA preparation, sequenced using the Illumina paired-end sequencing technology and assembled into unigenes. These unigenes were then analyzed in their genome mapping, functional annotation, homology, codon usage bias and simple sequence repeats (SSRs). RESULTS: Approximately 51.6 million clean reads were obtained, trimmed, and assembled into 38,504 unigenes with an average length of 571 bp, an N50 of 711 bp, and an average GC content 51.26%. Among them, 98.4% of unigenes could be mapped onto the reference genome, and 69% of unigenes could be annotated with known biological functions. Homology analysis identified certain numbers of An. sinensis unigenes that showed homology or being putative 1:1 orthologues with genomes of other Dipteran species. Codon usage bias was analyzed and 1,904 SSRs were detected, which will provide effective molecular markers for the population genetics of this species. CONCLUSIONS: Our data and analysis provide the most comprehensive transcriptomic resource and characteristics currently available for An. sinensis, and will facilitate genetic, genomic studies, and further vector control of An. sinensis.