Q-PCR primer, identification method and kit for identifying schistosoma japonicum infected oncomelania

A technology of schistosomiasis and kits, applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of poor timeliness, easy identification errors, and low accuracy, and achieve accurate results, Avoid time-consuming and labor-intensive, easy-to-operate effects

Active Publication Date: 2015-05-27
JIANGSU INST OF PARASITIC DISEASES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has high requirements for technicians, and the accuracy rate is low, and identification errors are prone to occur. After the snails are infected, due to the time required for the development of schistosomes, the earliest time for microscopic examination is 9 days, especially for oncomelania collected on site. After a period of time in the laboratory, it can be judged by a microscope. The timeliness is too poor to meet the current needs of schistosomiasis prevention work.

Method used

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  • Q-PCR primer, identification method and kit for identifying schistosoma japonicum infected oncomelania
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  • Q-PCR primer, identification method and kit for identifying schistosoma japonicum infected oncomelania

Examples

Experimental program
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Embodiment 1

[0022] Example 1 Identification method for Schistosoma japonicum-infected snails.

[0023] A total of 100 Oncomelania snails were raised in the laboratory, including 50 artificially infected snails and 50 uninfected controls. Total RNA was extracted using Trizol combined with RNA extraction kit;

[0024] A pair of Q-PCR primers were rationally designed using the A gene sequence of Schistosoma japonicum, and the sequence of the Q-PCR primers was:

[0025] Upstream primer PF: 5'-GGTCCATGTTTGGGTGGAGT-3'

[0026] Downstream primer PR: 5'-ATTCGGGTGTTCTTGAGGCT-3.

[0027] Wherein, the sequence of the A gene is shown in SEQNO1, which is cloned on insects by designing primers according to the human gene sequence, and the primers used for cloning are:

[0028] Upstream primer PF: 5'-tcagAAGCTT ATGGGGCGTACTGATACATTTG-3';

[0029] Downstream primer PR: 5'-actg AGATCT TTATAATCCCCGAGTTAGTAAG-3';

[0030] The length of the cloned product is 950bp, and it has been verified that the sequ...

Embodiment 2

[0036] Collect 10 river beach snails from the coast of the Yangtze River and 10 mountain snails from mountainous areas, and use Trizol RNA extraction kit to extract total RNA;

[0037] A pair of Q-PCR primers were rationally designed using the A gene sequence of Schistosoma japonicum, and the sequence of the Q-PCR primers was:

[0038] Upstream primer PF: 5'-GGTCCATGTTTGGGTGGAGT-3'

[0039] Downstream primer PR: 5'-ATTCGGGTGTTCTTGAGGCT-3.

[0040] Wherein, the sequence of the A gene is shown in SEQNO1, which is cloned on insects by designing primers according to the human gene sequence, and the primers used for cloning are:

[0041] Upstream primer PF: 5'-tcagAAGCTT ATGGGGCGTACTGATACATTTG-3';

[0042] Downstream primer PR: 5'-actg AGATCT TTATAATCCCCGAGTTAGTAAG-3';

[0043] The length of the cloned product is 950bp, and it has been verified that the sequence does exist and is correct.

[0044] Using the kit (ThermoScript TM RT-PCR System, Cat no.11146-024, including trizo...

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Abstract

The invention a Q.PCR primer for identifying schistosoma japonicum infected oncomelania, and also provides a method for identifying the schistosoma japonicum infected oncomelania by use of the primer. The PCR primer is reasonably designed according to the special A gene sequence of the schistosoma japonicum, and the PCR primer is combined with reverse transcription, PCR amplification and amplification curve interpretation to identify whether the oncomelania is of positive infection of the schistosoma japonicum; the identification method has the advantages of simple operation, high sequencing, quick identification, accurate result and the like; the defects of high time and labor consumption, poor repeatability, large man-made interference factor and the like of a traditional microscopic dissection and identification method are avoided.

Description

technical field [0001] The invention relates to the field of molecular biology detection, and specifically relates to Q-PCR primers, an identification method and a kit for identifying snails infected by Schistosoma japonicum. Background technique [0002] Schistosomiasis japonicum is a zoonotic parasitic disease that seriously endangers human health. Prevention of schistosomiasis infection is one of the most important public health tasks. According to the 2011 national schistosomiasis epidemic situation report, there are 286,836 schistosomiasis patients nationwide , including 30,028 patients with advanced schistosomiasis. Effective prevention and control of schistosomiasis infection in epidemic areas is crucial to economic and social stability. [0003] Oncomelania is the only intermediate host of Schistosoma japonicum, and it is widely distributed in the Yangtze River Basin of my country, especially in Hunan, Hubei, Jiangxi, Anhui, Jiangsu and other provinces. The transmis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/6888C12Q2531/113C12Q2563/107
Inventor 黄玉政刘坤
Owner JIANGSU INST OF PARASITIC DISEASES
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