An integrated flow chip for identifying fruit fly species and its application
A fruit fly and chip technology, which is used in the determination/inspection of microorganisms, DNA/RNA fragments, chemical libraries, etc., and can solve the problems of insect molecular identification applications that have not been reported.
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Embodiment 1
[0130] Embodiment 1, identification of 27 kinds of fruit fly specific primer pairs and the acquisition of probes
[0131] 1. Acquisition of the fruit fly mtDNA COI gene sequence
[0132] 1. The fruit fly samples to be tested are shown in Table 1. The samples were all placed in absolute ethanol and stored at -20°C for later use.
[0133] Table 1. Types of fruit fly samples tested and information on collection places
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[0138] 2. Use the "Blood / Cell / Tissue Genomic DNA Extraction Kit" (Tiangen Biochemical Technology (Beijing) Co., Ltd.) to extract the genomic DNA of each fruit fly sample in Table 1 (most samples are extracted from a single foot, and a few The sample is extracted from a single head), and the specific operation process refers to the instructions to obtain the genomic DNA of each fruit fly sample in Table 1.
[0139] The genomic DNA of each fruit fly sample in Table 1 was used as a template, and the universal primer L...
Embodiment 2
[0150] Example 2, Preparation of integrated flow chip for identification of 27 species of fruit flies and its specificity detection
[0151] 1. Preparation of integrated flow chip
[0152] 1. Initialization of 48.48 dynamic chip
[0153] (1) Open the package of the chip and use it within 24 hours;
[0154] (2) Inject the Control Line Fluid into the collectors at both ends of the 48.48 dynamic chip, and be careful not to drop the Control Line Fluid into the Assay Inlets or Sample Inlets;
[0155] (3) Remove the blue protective film on the bottom of the IFC chip;
[0156] (4) Transfer the chip to the IFC Controller, run the Primer (113x) program to initialize the chip, and the initialization time is 10 minutes; after the initialization is completed, the sample loading operation is completed within 60 minutes.
[0157] 2. Preparation of standard reaction system and sample mixture
[0158] (1) According to the test system shown in Table 3, prepare 10×Assays of 27 kinds of frui...
Embodiment 3
[0210] Example 3, the application of integrated flow chip in the identification of fruit flies
[0211] 1. Fruit fly samples to be tested
[0212] Taking the following non-adult fruit fly sample as an example, the integrated flow chip described in Example 2 was used to identify its species.
[0213] In May 2012, Beijing Entry-Exit Inspection and Quarantine Bureau intercepted 3 fruit fly larvae from lotus mist imported from Thailand;
[0214] In May 2014, Beijing Entry-Exit Inspection and Quarantine Bureau intercepted 2 fruit fly larvae from mangoes imported from Tanzania;
[0215] In July 2009, two fruit fly larvae were collected from the rotten fruits of bitter gourd in Nanning, Guangxi Zhuang Autonomous Region, my country;
[0216] In October 2011, two fruit fly pupae whose host was citrus were submitted to the Plant Protection and Inspection Station of Chengdu City, Sichuan Province, my country;
[0217] In August 2011, 3 capsicum fruit fly eggs were raised in the Nation...
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