tetano
Editor, Senior Moderator
Spectrochim Acta A Mol Biomol Spectrosc
. 2025 Feb 21:335:125943.
doi: 10.1016/j.saa.2025.125943. Online ahead of print. A programmable nucleic acid fluorescence biosensor based on the BstNI endonuclease for detection of the influenza A (H1N1) virus
Qila Sa[SUP] 1 [/SUP], He Sun[SUP] 2 [/SUP], Shengjun Bu[SUP] 2 [/SUP], Yao Xu[SUP] 2 [/SUP], Zebin Zhang[SUP] 1 [/SUP], Chenxi Li[SUP] 1 [/SUP], Longtao Wang[SUP] 3 [/SUP], Jiayu Wan[SUP] 4 [/SUP]
Affiliations
Timely and accurate diagnosis of influenza virus is essential to prevent the spread of disease and to select an appropriate treatment strategy. Here, we report the development of a novel fluorescence biosensor for detection of the H1N1 virus based on the BstNI endonuclease, dually-blocked RNA strands (S), and FAM-ssDNA-Q reporter (R) strands. The S strand contains a short (5 nt) sequence for BstNI recognition and sequences on both sides of the cutting site, which are closed by two locking strands. The R strand is complementary to the intermediate sequence of the S strand, including the BstNI cutting site domain and partial adjacent sequences. Only the presence of two specific RNA fragments of the target influenza virus can fully de-block the S strands, which then hybridize with the R strands, followed by cleavage via BstNI-catalyzed hydrolysis, thereby generating the fluorescence signal. The biosensor was sensitive to the H1N1 virus at 100 fM and was highly specific to the target sequence. The proposed biosensor provides a convenient method for reliable detection of the H1N1 virus.
Keywords: BstNI endonuclease; Fluorescent biosensor; H1N1; Primer exchange reaction.
. 2025 Feb 21:335:125943.
doi: 10.1016/j.saa.2025.125943. Online ahead of print. A programmable nucleic acid fluorescence biosensor based on the BstNI endonuclease for detection of the influenza A (H1N1) virus
Qila Sa[SUP] 1 [/SUP], He Sun[SUP] 2 [/SUP], Shengjun Bu[SUP] 2 [/SUP], Yao Xu[SUP] 2 [/SUP], Zebin Zhang[SUP] 1 [/SUP], Chenxi Li[SUP] 1 [/SUP], Longtao Wang[SUP] 3 [/SUP], Jiayu Wan[SUP] 4 [/SUP]
Affiliations
- PMID: 40023959
- DOI: 10.1016/j.saa.2025.125943
Timely and accurate diagnosis of influenza virus is essential to prevent the spread of disease and to select an appropriate treatment strategy. Here, we report the development of a novel fluorescence biosensor for detection of the H1N1 virus based on the BstNI endonuclease, dually-blocked RNA strands (S), and FAM-ssDNA-Q reporter (R) strands. The S strand contains a short (5 nt) sequence for BstNI recognition and sequences on both sides of the cutting site, which are closed by two locking strands. The R strand is complementary to the intermediate sequence of the S strand, including the BstNI cutting site domain and partial adjacent sequences. Only the presence of two specific RNA fragments of the target influenza virus can fully de-block the S strands, which then hybridize with the R strands, followed by cleavage via BstNI-catalyzed hydrolysis, thereby generating the fluorescence signal. The biosensor was sensitive to the H1N1 virus at 100 fM and was highly specific to the target sequence. The proposed biosensor provides a convenient method for reliable detection of the H1N1 virus.
Keywords: BstNI endonuclease; Fluorescent biosensor; H1N1; Primer exchange reaction.