Title:
EXTENDED PROBE-TYPE QPCR DETECTION METHOD FOR SNV
Document Type and Number:
WIPO Patent Application WO/2023/030162
Kind Code:
A1
Abstract:
Disclosed in the present invention is an extended probe-type qPCR detection method for single nucleotide variation (SNV). Four primers are designed according to known single nucleotide variation sites, with two of the primers being external primers and the other two being specific primers, wherein bases at the 3' ends of the specific primers are respectively complementary with bases of a mutation template and a normal template; mismatches can be introduced or not introduced into bases near the 3' ends; and a segment of artificial sequence is extended at the 5' end of each specific primer, with an accessory primer in the same direction designed on the 5' segment of the artificial sequence and an accessory probe designed on the 3' segment thereof. In the amplification of the specific primers and downstream primers, the accessory probes on the specific primers are degraded and emit light in the extension of the accessory primers so as to indicate the amplification efficiency of the specific primers. If only one genotype of the single nucleotide variation sites is detected, two external primers, one specific primer, and the accessory primer and probe of the specific primer can be used. The method still has many implementation modes, and has a higher sensitivity and a lower cost than a probe-type AS-PCR.
Inventors:
YANG YONGCHEN (CN)
ZHANG HONG (CN)
ZHANG HONG (CN)
Application Number:
PCT/CN2022/114842
Publication Date:
March 09, 2023
Filing Date:
August 25, 2022
Export Citation:
Assignee:
CHILDRENS HOSPITAL OF SHANGHAI (CN)
DAOZHI PRECISION MEDICINE TECH SHANGHAI CO LTD (CN)
DAOZHI PRECISION MEDICINE TECH SHANGHAI CO LTD (CN)
International Classes:
C12Q1/6858; C12N15/11; C12Q1/70
Domestic Patent References:
| WO2005059178A1 | 2005-06-30 |
Foreign References:
| CN113846147A | 2021-12-28 | |||
| US20210207204A1 | 2021-07-08 | |||
| CN101354337A | 2009-01-28 | |||
| CN106520950A | 2017-03-22 | |||
| CN113308519A | 2021-08-27 | |||
| JP2007330135A | 2007-12-27 | |||
| CN109136345A | 2019-01-04 |
Attorney, Agent or Firm:
DAOZHI PRECISION MEDICINE TECHNOLOGY SHANGHAI CO LTD (CN)
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