Primers, probes and kits for simultaneous detection of human herpesvirus types 6 and 7
A technology for simultaneous detection of human herpes virus, which is applied in the field of probes and kits, type 7 primers, and simultaneous detection of human herpes virus 6, can solve the problems of high false positive rate, achieve good specificity, reduce false positive rate, The effect of high sensitivity
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Embodiment 1
[0033] The present embodiment provides a set of primers and fluorescent probes for simultaneous detection of human herpesvirus 6 and 7. The reorganization contains two pairs of primers and two probes. The sequence is as follows:
[0034] (1) Forward primer HHV6-F, the nucleotide sequence of which is shown in SEQ ID NO.1;
[0035] Reverse primer HHV6-R, the nucleotide sequence of which is shown in SEQ ID NO.2;
[0036] Fluorescent probe HHV6-P, the nucleotide sequence of which is shown in SEQ ID NO.3, the 5' end of the fluorescent probe is labeled with FAM, and the 3' end quencher group BHQ;
[0037] (2) Forward primer HHV7-F, the nucleotide sequence of which is shown in SEQ ID NO.4;
[0038] Reverse primer HHV7-R, the nucleotide sequence of which is shown in SEQ ID NO.5;
[0039] Fluorescent probe HHV7-P, the nucleotide sequence of which is shown in SEQ ID NO.6, the 5' end of the fluorescent probe is labeled JOE, and the 3' end quencher group BHQ;
[0040] The above primers...
Embodiment 2
[0042] This embodiment provides a fluorescent quantitative PCR kit for simultaneous detection of human herpesvirus 6 and 7
[0043] 1. Kit composition and preparation
[0044] 1) Primer and fluorescent probe set
[0045] Same as Example 1
[0046] 2) DNA extraction solution
[0047] The DNA extraction solution can use a commercially available DNA extraction kit, or it can be prepared by itself.
[0048] In this example, the formula of the DNA extract solution: prepare 200mmol / L NaCl, 100mmol / L Tris-HCl, 1% SDS, 50mmol / L EDTA, adjust the pH to 8.5, 3mg / ml proteinase K (add before use)
[0049] 3) The reaction system of double fluorescent quantitative PCR is as follows:
[0050] Reagent Final concentration or content per unit volume PCR 10×buffer 5μl MgCl 2
4mmol / L dNTP (including dUTP) 0.48mmol / L Tag DNA polymerase 0.5U UNGase 0.2U HHV-6 primer 0.29μmol / L HHV-6 probe 0.24μmol / L HHV-7 primer 0.29μmol / L ...
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