Title:
METHOD AND DETECTION KIT FOR DETECTING ACTIVITY OF DNA POLYMERASE
Document Type and Number:
WIPO Patent Application WO/2018/205157
Kind Code:
A1
Abstract:
A method and a detection kit for detecting activity of a DNA polymerase. A DNA polymerase to be tested is mixed with a sufficient amount of a template to be filled, and in the presence of dNTP, a first DNA fragment in the template to be filled is induced by the DNA polymerase to be tested to form a filled fragment with bases complementary to bases of a protruding fragment, so as to obtain a filled template. The filled template is mixed with a forward primer, a reverse primer, a probe, dNTP, and a sufficient amount of UDG enzyme to carry out a real-time fluorescent quantitative PCR amplification reaction. A CT value of the PCR amplification reaction of the DNA polymerase to be tested is introduced into a standard curve created from a DNA polymerase standard, and the enzyme activity of the DNA polymerase to be tested can be calculated.
Inventors:
CAI TONGCONG (CN)
DENG YANHUA (CN)
YANG HAO (CN)
HUANG TIANTIAN (CN)
SUN KANGCHENG (CN)
WANG YIQIONG (CN)
DENG YANHUA (CN)
YANG HAO (CN)
HUANG TIANTIAN (CN)
SUN KANGCHENG (CN)
WANG YIQIONG (CN)
Application Number:
PCT/CN2017/083690
Publication Date:
November 15, 2018
Filing Date:
May 10, 2017
Export Citation:
Assignee:
FAPON BIOTECH INC (CN)
GUANGDONG FAPON BIOTECH CO LTD (CN)
GUANGDONG FAPON BIOTECH CO LTD (CN)
International Classes:
C12Q1/68; C12Q1/48
Foreign References:
| CN104357543A | 2015-02-18 | |||
| CN104313132A | 2015-01-28 | |||
| CN102534036A | 2012-07-04 |
Other References:
ZWEITZIG, D.R. ET AL.: "Characterization of a Novel DNA Polymerise Activity Assay Enabling Sensitive, Quantitative and Universal Detection of Viable Microbes", NUCLEIC ACIDS RESEARCH, vol. 40, no. 14, 4 November 2012 (2012-11-04), pages 1 - 12, XP055106744
MA, C.B. ET AL.: "Label-free Molecular Beacon for Real-time Monitoring of DNA Polymerase Activity", ANAL BIOANAL CHEM., vol. 408, no. 12, May 2016 (2016-05-01), pages 3275 - 3280, XP055545903
YU , L.M. ET AL.: "Fluorescence-Based, High- Throughput DNA Polymerase Assay", BIOTECHNIQUES, vol. 33, no. 4, 31 October 2002 (2002-10-31), pages 938 - 941, XP055545906
ZWEITZIG, D.R. ET AL.: "A Novel Approach for Rapid Detection of Bacterially Contaminated Platelet Concentrates via Sensitive Measurement of Microbial DNA Polymerase Activity", TRANSFUSION, vol. 54, no. 6, 1 July 2014 (2014-07-01), pages 1642 - 1651, XP055545911
MA, C.B. ET AL.: "Label-free Molecular Beacon for Real-time Monitoring of DNA Polymerase Activity", ANAL BIOANAL CHEM., vol. 408, no. 12, May 2016 (2016-05-01), pages 3275 - 3280, XP055545903
YU , L.M. ET AL.: "Fluorescence-Based, High- Throughput DNA Polymerase Assay", BIOTECHNIQUES, vol. 33, no. 4, 31 October 2002 (2002-10-31), pages 938 - 941, XP055545906
ZWEITZIG, D.R. ET AL.: "A Novel Approach for Rapid Detection of Bacterially Contaminated Platelet Concentrates via Sensitive Measurement of Microbial DNA Polymerase Activity", TRANSFUSION, vol. 54, no. 6, 1 July 2014 (2014-07-01), pages 1642 - 1651, XP055545911
Attorney, Agent or Firm:
ADVANCE CHINA IP LAW OFFICE (CN)
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