RT-RPA kit, primer and probe for visually and rapidly detecting Chinese softshell turtle hemorrhagic syndrome virus TSHSV
A RT-RPA, syndrome virus technology, applied in the field of biological detection, can solve the problems of inability to judge samples, confusion of disease samples, inability to diagnose, etc., and achieve the effects of clear visual detection, specific detection results, and short detection time.
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Embodiment 1
[0081] An RT-RPA kit for visual and rapid detection of Chinese soft-shelled turtle hemorrhagic syndrome virus TSHSV, said kit comprising: a dry powder reaction tube containing RPA freeze-dried powder; said RPA freeze-dried powder includes recombinant enzyme, single-stranded DNA binding protein , DNA polymerase, auxiliary protein and M-MLV reverse transcriptase (both enzyme and protein are from Weifang Anpu Future Biotechnology Co., Ltd.);
[0082] Buffer A, the composition of the buffer A is: 6wt% polyethylene glycol, Tris-HCL 30mM, ddH 2 O margin;
[0083] Buffer B, the buffer B is a magnesium acetate solution with a concentration of 280mM;
[0084] Primer, the sequence of the primer is:
[0085] Upstream primer TSHSV-F2: 5'-AAGCTTCCAGCCTATTGGCCTCCATTTACC-3'
[0086] Downstream primer TSHSV-R1: [5'-biotin]AATTGTCTTTCCCAACATACTCACTTCCGTTG-3';
[0087] A colloidal gold probe, the colloidal gold probe is TSHSV-nfo-PB, and its sequence is [5'-6FAM]TCTTGTGCTGGATGTTGGCGAATTCTTG...
Embodiment 2
[0090] The difference between this embodiment and Example 1 is that the sequence of the primer is:
[0091] Upstream primer TSHSV-F4: 5'-TTGTTGAGCAAGACCACCAAGGACGACCCT-3'
[0092] Downstream primer TSHSV-R2: [5'-biotin]AATCTCAGAGGTCAGCTTCTCAATAGTTTTG-3';
[0093] Others are with embodiment 1.
Embodiment 3
[0095]An RT-RPA kit for visual and rapid detection of Chinese soft-shelled turtle hemorrhagic syndrome virus TSHSV, said kit comprising: a dry powder reaction tube containing RPA freeze-dried powder; said RPA freeze-dried powder includes recombinant enzyme, single-stranded DNA binding protein , DNA polymerase, auxiliary protein and M-MLV reverse transcriptase (both enzyme and protein are from Weifang Anpu Future Biotechnology Co., Ltd.);
[0096] Buffer A, the composition of the buffer A is: 6wt% polyethylene glycol, Tris-HCL 30mM, ddH 2 O margin;
[0097] Buffer B, the buffer B is a magnesium acetate solution with a concentration of 280mM;
[0098] Primer, the sequence of the primer is:
[0099] Upstream primer TSHSV-F2: 5'-AAGCTTCCAGCCTATTGGCCTCCATTTACC-3'
[0100] Downstream primer TSHSV-R1: [5'-biotin]AATTGTCTTTCCCAACATACTCACTTCCGTTG-3';
[0101] A fluorescent probe, the fluorescent probe being TSHSV-exo-PB,
[0102] Its sequence is: 5'-TCTTGGCCTTGAACCAGTATACTGTTACAAA...
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