Assays to distinguish between tick species and intraspecies specificity
A specific, blood tick technology, applied in biochemical equipment and methods, microbial determination/inspection, sequence analysis, etc., can solve the problems of complex identification process, difficult morphological identification and distinction, etc.
Active Publication Date: 2021-08-20
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology
For the morphological identification of ticks, experts with extensive experience and knowledge are required, and the identification process is very complicated
In addition, for immature individuals (eggs, young ticks) and incomplete specimens, it is difficult to distinguish through morphological identification (Deng Guofan, Jiang Zaijie. Chinese Economic Entomology [J]. Volume 39 - Acari Ixodidae, Beijing = Science Press, 1991.)
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[0033] (1) Tick genome extraction
[0034] Preparation before the experiment: Use the (QIAGEN) kit to extract the experimental tick genome, see the following steps:
[0035] 1. Place the experimental ticks at room temperature (15°C-25°C);
[0036] 2. Prepare two temperature bath systems: step 3 needs to be placed at 56°C; step 5 needs to be placed at 70°C;
[0037] 3. The AE buffer or sterilized water required in step 11 should be placed at room temperature;
[0038] 4. Confirm that AW1 and AW2 (AW1 and AW2 are the two washing buffers in the QINGEN kit are concentrated, so it is necessary to confirm whether the specified amount of 96%-100% ethanol has been added before the first use). process as requested;
[0039] 5. If there is precipitation in BufferAE or BufferAL (the tick genome collection buffer provided in the QIAGEN kit), or crystallization may occur at low temperature due to temperature reasons, it can be dissolved by heating at 56°C.
[0040] The specific operati...
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The invention discloses a method for identifying and distinguishing different tick species and the same tick species in different regions simply, quickly and accurately. The method of the present invention is as follows: extract the genomic DNA of ticks, and then amplify with corresponding SCAR primers, electrophoresis the amplified products in the gel, and determine the species of ticks to be tested according to the bands appearing at specific positions in the electrophoresis graph. The preparation method of the SCAR primer used in the method of the present invention is, with reference to the common SRAP primer sequence of species, find out the sequence amplification polymorphism fragment SRAP relevant with tick from the tick genome, then find out sequence characteristic from the tick SRAP fragment Amplify the region (SCAR) fragment, then design and obtain corresponding tick SCAR amplification primers. The method of the invention has strong specificity, good repeatability and high accuracy, and can be widely applied to the specific identification of tick species and the precise tracing of the epidemic lineage within a species.
Description
technical field [0001] The present invention relates to a detection and identification technology for ticks. Specifically, it is a simple, fast and accurate method for identifying and distinguishing different tick species and different regions for the same tick species, while providing inter-species and intra-species detection of ticks. Differential test kits. Background technique [0002] Ticks belong to Arthropoda (phylum Arthropoda), subclass Acari (Parasitiformes), subclass Parasitiformes (Ixodida), including Ixodidae (Ixodidae), soft ticks There are more than 900 species in 3 families, Argasidae and Nuttalliellidae (Yang Ya, Liu Jinhui, Fu Jian, et al. Research progress on molecular taxonomy of hard ticks[J]. Advances in Veterinary Medicine, 2014, 35(11) : 98-100; Barker S C, Murrell A. Systematics and evolution ofticks with a list of valid genus and speciesnames [J]. Parasitology, 2004, 129: 15-36.), is a kind of obligate external blood-sucking parasite. At present, ...
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IPC IPC(8): C12Q1/6888C12Q1/6858C12N15/11
CPCC12Q1/6888G16B30/00
Inventor 刘光远刘小翠罗金陈泽任巧云刘旻翾刘志杰独军政关贵全罗建勋殷宏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI