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Geographically-specific Plasmodium vivax molecule marker, and its application in strain tracing

A molecular marker, Plasmodium technology, applied in the field of biotechnology and molecular biology, can solve the problem of difficult to fully reflect the genetic characteristics of the strain population

Pending Publication Date: 2014-06-18
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main reason is that population segregation is affected by multiple factors such as natural selection, genetic drift, migration, mutation, and isolation, and it is often difficult for a single index to fully reflect the population genetic characteristics of strains in endemic areas.

Method used

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  • Geographically-specific Plasmodium vivax molecule marker, and its application in strain tracing
  • Geographically-specific Plasmodium vivax molecule marker, and its application in strain tracing
  • Geographically-specific Plasmodium vivax molecule marker, and its application in strain tracing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0139] Classification of core nonapeptide repeats in the central repeat region of the circumsporozoite protein of Plasmodium vivax, and creation of a characterization model

[0140] 1. Detection of the core nonapeptide repeat sequence in the central repeat region of the circumsporozoite protein of Plasmodium vivax

[0141] (a) Design of primers

[0142] The sequence is from the reference sequence of Plasmodium vivax Belem strain (see SEQ ID NO: 1 for the full sequence, GenBank accession no. M 11926).

[0143] The outer primers for amplifying the central repeat region are:

[0144] CSP-WF: 5'GCATAAGGCAAACTCACAAA 3' (SEQ ID NO: 4),

[0145] CSP-WR: 5'CGCATAATGTGTAAGAGGTGT 3' (SEQ ID NO: 5).

[0146] The inner primer is: T-CS-F:5'AAGCAAGCAAAACAGCCAAA 3'(SEQ ID NO:6),

[0147] T-CS-R: 5'GCAAACAGCAGGGAACATT 3' (SEQ ID NO: 7).

[0148] (b) PCR amplification conditions are: 98° C. for 20 seconds (denaturation), 57° C. for 30 seconds (annealing) and 68° C. for 60 seconds (extensi...

Embodiment 2

[0183] Classification of core SNPs in dihydrofolate reductase and dihydropterin synthase, and creation of signature models

[0184] 1. Detection of Plasmodium vivax dihydrofolate reductase and dihydropterin synthase

[0185] (a) Design of primers

[0186] The sequences were obtained from the reference sequence of Plasmodium vivax Sal-I strain (full sequences are respectively SEQ ID NO: 2 and SEQ ID NO: 3, GenBank accession no. X98123 and AY 186730).

[0187] The outer primers for amplifying DHFR are:

[0188] POF: 5'CACCGCACCAGTTGATTCCT 3' (SEQ ID NO: 8),

[0189] POR: 5' CCTCGGCGTTGTTCTTCT 3' (SEQ ID NO: 9).

[0190] The inner primer is: PBF:5'CCCCACCACATAACGAAG 3'(SEQ ID NO:10),

[0191] PAR: 5' CCCCACCTTGCTGTAAAC C 3' (SEQ ID NO: 11).

[0192] The outer primers for amplifying DHPS are:

[0193] DHPS-F1: 5'GATGGCGGTTTATTTGTCG 3' (SEQ ID NO: 12),

[0194] DHPS-R1: 5'GCTGATCTTTGTCTTGACG 3' (SEQ ID NO: 13).

[0195] The inner primers are:

[0196] DHPS-F2: 5'GCTGTGGAGA...

Embodiment 3

[0217] Create a taxonomic model featuring core-neutral microsatellite loci

[0218] 1. Detection of neutral microsatellites

[0219] (a) Design of primers

[0220] The primers for amplifying the 3.35 site are:

[0221] F: 5'TGAGAGGAGCCTACTGTGAT 3' (SEQ ID NO: 16),

[0222] R: 5'GCCACAGGATGTACATAAGA 3' (SEQ ID NO: 17),

[0223] F2: 5'CCAAGTAGAGAAAGGGAAAA 3' (SEQ ID NO: 18).

[0224] The primers for amplifying the 9-AT locus are:

[0225] F: 5'TGCTTGTGCGTATGCTGC 3' (SEQ ID NO: 19),

[0226] R: 5'CCGCCTGGTTGACGTTTC 3' (SEQ ID NO: 20),

[0227] F2: 5'TCCTCGGCTTACAAACGC 3' (SEQ ID NO: 21).

[0228] The primers for amplifying the 12.335 site are:

[0229] F: 5'TTAGTTCCAGCAAAACCTTC 3' (SEQ ID NO: 22),

[0230] R: 5'TTATAACCTTCGGGGTTTTT 3' (SEQ ID NO: 23),

[0231] F2: 5'AAAAATGGAGACATGGAAGA 3' (SEQ ID NO: 24).

[0232] (b) PCR amplification conditions are: 98° C. for 20 seconds (denaturation), 52° C. for 30 seconds (annealing) and 68° C. for 30 seconds (extension), 20 cycles...

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Abstract

The invention relates to a geographically-specific Plasmodium vivax molecule marker, and its application in strain tracing. The molecule marker includes a Plasmodium vivax circumsporozoite protein center replication region reflecting mosquito infection differences of different media, a height polymorphism microsatellite reflecting mutation, migration and genetic drift, and a drug resistance related gene mutation reflecting the population positivity selection of drug control. Results show that each of a Plasmodium vivax circumsporozoite protein center replication region sequence, dihydrofolate reductase SNP, dihydrobiopterin synthetase SNP, a neutral microsatellite and the like has a geographic specificity, and can be used as a classification index in tracing detection.

Description

technical field [0001] The invention belongs to the field of biotechnology and molecular biology, in particular, the invention relates to a group of geographically specific Plasmodium vivax molecular markers and their application in traceability of strains. Background technique [0002] Malaria is one of the oldest infectious diseases among human diseases, and it still seriously affects human health today. According to the World Malaria Report 2011 released by the World Health Organization (WHO), there are approximately 216 million malaria cases and 655,000 deaths worldwide each year. Although the malaria mortality rate has dropped by 25% in the past decade, factors such as the spread of drug-resistant plasmodium, frequent personnel exchanges, and global warming are threatening the hard-won achievements in anti-malaria. [0003] In my country, the 40 years of malaria prevention and control work has achieved remarkable results, the scope of endemic areas has been greatly red...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6893C12Q2600/156G01N33/6818G01N2333/445Y02A50/30
Inventor 潘卫庆丁帅张冬梅
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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