Papillomavirus detection and parting method as well as liquid phase chip thereof
A technology of virus detection and liquid phase chip, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of poor signal-to-noise ratio of detection results, increase coating steps, limit sensitivity, etc., to achieve increased sensitivity, The effect of increasing the fluorescence signal value and improving the detection efficiency
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Embodiment 1
[0026] Embodiment 1 Human papillomavirus HPV detection and typing liquid phase chip kit mainly includes:
[0027] 1. Specific primer sequence (ASPE primer)
[0028]For 22 types of HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 70, 73, 82, 06, 11, 42, 43, 44 Design specific primer sequences (18-22bp), add a 24bp specific tag tag sequence to the 5' end of the primers, and these tag tag sequences can specifically complement each other with the corresponding anti-tag sequences on the microspheres. The ASPE sequence is shown in the table below:
[0029] Table 1 ASPE primer sequence
[0030] SEQ NO.
type
NCBI serial number
Sequence(5'→3')
[0031] 1
HPV06
X00203
AATCTACAAATCCAATAATCTCATTCCGTAACTACATCTTCCA
2
HPV11
M14119
CAATTCAAATCACAATAATCAATCTCTGTGTCTAAATCTGCTAC
3
HPV16
K02718
CAATTCATTTCATTCACAATCAATTACCCTACGACATGGGGAG
4
HPV18
X05015
TCAACAATCTTTTACAATC...
Embodiment 2
[0045] Example 2 Detection and typing of clinical samples using HPV detection and typing liquid chip
[0046] The formula of described various solutions is as follows:
[0047] 50mM MES buffer (pH5.0) formula (250ml):
[0048] Reagent
source
Final concentration
Dosage per 250ml
[0049] MES
(2[N-Morpholino]
ethanesulfonic
acid)
Sigma M-2933
0.05M
2.44g
5MNaOH
Fisher SS256-500
---
5 drops
[0050] 2×Tm hybridization buffer
[0051] Reagent
source
Final concentration
Dosage per 250ml
1M Tris-HCl, pH8.0
SigmaT3038
0.2M
50ml
5M NaCl
Sigma S5150
0.4M
20ml
Triton X-100
Sigma T8787
0.16%
0.4ml
[0052] Store at 4°C after filtration
[0053] Biotin-labeled dNTPs were purchased from Shanghai Sangon Bioengineering Technology Service Co., Ltd.
[0054] 1. Sample selec...
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