Multiple PCR primer for expanding plasmodium falciparum drug resistance-related gene and method

A technology related to Plasmodium falciparum and resistance, applied in the field of nucleotide determination or inspection, can solve the problems of cumbersome operation, time-consuming and labor-consuming, etc., achieve fewer operation steps, reduce the possibility of operation errors, and improve the success of experiments The effect of chance

Active Publication Date: 2008-04-16
STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
View PDF0 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0149] Aiming at the disadvantages of cumbersome operation, time-consuming and labor-intensive traditional amplification methods of Plasmodium falciparum drug resistance-related genes, the present invention provides an efficient and convenient multiplex PCR amplification primer and method for Plasmodium falciparum drug resistance-related genes, realizing one-time The reaction completed the amplification of 5 main resistance-related genes (respectively reflecting the resistance of Plasmodium falcipar

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Multiple PCR primer for expanding plasmodium falciparum drug resistance-related gene and method
  • Multiple PCR primer for expanding plasmodium falciparum drug resistance-related gene and method
  • Multiple PCR primer for expanding plasmodium falciparum drug resistance-related gene and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0183] (1) Primers

[0184] According to the conserved sequences of 5 Plasmodium falciparum genes Pfcrt, Pfmdr1, Pfdhps, Pfdhfr and PfATPase6 and the positions of 21 drug resistance-related SNP sites, 5 pairs of specific primers were designed and synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd. . The expected lengths of the amplified products of the five genes were Pfcrt: 315bp; Pfmdr1: 514bp; Pfdhps: 770bp; Pfdhfr: 594bp; PfTPase6: 437bp.

[0185] Pfcrt

[0186] Upstream primer (P 1F): 5'-GGAGGTTCTTGTCTTGGTAAAT-3'

[0187] Downstream primer (P1R): ​​5'-ATATTGGTAGGTGGAATAGATTCT-3'

[0188] Pfmdr1

[0189] Upstream primer (P2F): 5'-TGTTGAAAGATGGGTAAAGAGCAGA-3'

[0190] Downstream primer (P2R): 5'-TCGTACCAATTCCTGAACTCACTT-3'

[0191] Pfdhps

[0192] Upstream primer (P3F): 5'-GATTCTTTTTCAGATGGAGG-3'

[0193] Downstream primer (P3R): 5'-TTCCTCATGTAATTCATCTGA-3'

[0194] Pfdhfr

[0195] Upstream primer (P4F): 5'-TGATGGAACAAGTCTGCGACGTT-3'

[019...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to a plasmodium falciparum drug resistance related gene multiple PCR amplification method, wherein, plasmodium falciparum genome DNA is adopted as template; a multiple PCR primer set designed by the invention is adopted for monotube PCR reaction to realize amplification of five specific gene (plasmodium falciparum chloroquine resistance transport protein gene Pfcrt, plasmodium falciparum multidrug-resistance gene Pfmdrl, plasmodium falciparum dihydropteroic synthetase gene Pfdhps, plasmodium falciparum dihyrofolate reductase gene Pfdhfr and plasmodium falciparum adenosine triphosphatase Sixth subunit gene PfATPase6) target sequences; moreover, the amplification product covers the currently known 21 plasmodium falciparum drug-resistance related SNP sites. With fewer operational procedures, the invention is quick and convenient and saves cost; moreover, the invention which is particularly suitable for high-throughput genotype analysis is sure to accelerate the wide application of plasmodium falciparum resistance related SNP in field monitoring.

Description

technical field [0001] The invention relates to an amplification technology of a gene related to drug resistance of Plasmodium falciparum, and belongs to the technical field of nucleotide determination or detection methods. Background technique [0002] The prevalence of malaria is severe worldwide, especially the harm of falciparum malaria. According to the report of the World Health Organization, about 1 million people die of falciparum malaria every year in the world, not including indirect deaths. Falciparum malaria is prevalent in Yunnan and Hainan provinces in my country, and deaths are reported every year. After the 1950s, Plasmodium falciparum began to develop resistance to antimalarial drugs. In recent years, the resistance situation has become increasingly severe, from low resistance to high resistance, from single resistance to multiple resistance, and the speed of resistance development Gradually speed up. Accurate and rapid monitoring of the distribution and p...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68C12P19/34
CPCY02A50/30
Inventor 张国庆汤林华
Owner STATION OF VIRUS PREVENTION & CONTROL CHINA DISEASES PREVENTION & CONTROL CENT
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap