An amplification nanoswitch system based on split site-specific cleaving enzymes for the in vitro detection of target analytes, said amplification nanoswitch system comprising: - at least two catalytically inactive nanoswitch modules (P1, P2) each comprising: - at least one split catalytically inactive site-specific cleaving enzyme fragment, wherein said enzyme is a protease; - at least one binding moiety (BM) which is capable to bind a target analyte; - at least one binding arm configured to bind the at least one split catalytically inactive site-specific cleaving enzyme fragment to the at least one binding moiety (BM); - at least one effector molecule that contains a cleavage recognition site and a sensor or reporter molecule; - an enhancer module configured to increase signal intensity by increasing the enzymatic activity of the amplification nanoswitch system when the binding moieties bind the target analyte, said enhancer module comprising the following specific sub-modules: - at least two protease catalytically inactive fragments bound to one single stranded DNA; - at least on linking arm molecule bound to at least one protease catalytically inactive fragment capable to do self-anneal; - a peptide or chemical or nucleotide spacer containing at least two protease recognition sites. wherein said linking arm comprises, or is constituted by RNA, peptide sequences, DNA, PNA. - at least two catalytically inactive nanoswitch modules (P1, P2) each comprising: - at least one split catalytically inactive site-specific cleaving enzyme fragment, wherein said enzyme is a protease; - at least one binding moiety (BM) which is capable to bind a target analyte; - at least one binding arm configured to bind the at least one split catalytically inactive site-specific cleaving enzyme fragment to the at least one binding moiety (BM); - at least one split catalytically inactive site-specific cleaving enzyme fragment, wherein said enzyme is a protease; - at least one binding moiety (BM) which is capable to bind a target analyte; - at least one binding arm configured to bind the at least one split catalytically inactive site-specific cleaving enzyme fragment to the at least one binding moiety (BM); - at least one effector molecule that contains a cleavage recognition site and a sensor or reporter molecule; - an enhancer module configured to increase signal intensity by increasing the enzymatic activity of the amplification nanoswitch system when the binding moieties bind the target analyte, said enhancer module comprising the following specific sub-modules: - at least two protease catalytically inactive fragments bound to one single stranded DNA; - at least on linking arm molecule bound to at least one protease catalytically inactive fragment capable to do self-anneal; - a peptide or chemical or nucleotide spacer containing at least two protease recognition sites. wherein said linking arm comprises, or is constituted by RNA, peptide sequences, DNA, PNA. - at least two protease catalytically inactive fragments bound to one single stranded DNA; - at least on linking arm molecule bound to at least one protease catalytically inactive fragment capable to do self-anneal; - a peptide or chemical or nucleotide spacer containing at least two protease recognition sites. wherein said linking arm comprises, or is constituted by RNA, peptide sequences, DNA, PNA. <EOS>