Kit for detecting common depletion alpha-thalassemia and use method thereof
A technology of thalassemia and reagent kit, which is applied in the field of medical detection, and can solve problems such as long section length and long detection time
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Embodiment 1
[0072] Embodiment 1: The detection result of using the kit of the present invention in known genotype samples
[0073] 1. The composition of the kit:
[0074] (1) Primer pairs A1A2-F and A1A2-R that can simultaneously amplify the characteristic sequence A1 of the α1 segment and the characteristic sequence A2 of the α2 segment in the α-globin gene cluster:
[0075] A1A2-F: 5'-GGGTTGCGGGAGGTGTAG-3' (SEQ ID NO: 1);
[0076] A1A2-R: 5'-GCAGGCAGTGGCTTAGGA-3' (SEQ ID NO: 2);
[0077] (2) In the amplified α-globin gene cluster -- SEA Gap-PCR primer pair SEA-F and SEA-R for genotype:
[0078] SEA-F: 5'-CTCAGTATTGGAGGGAAGGA-3' (SEQ ID NO: 3);
[0079] SEA-R: 5'-CAGTGTTGTAGTCATGGCTTA-3' (SEQ ID NO: 4);
[0080] (3) In the amplified α-globin gene cluster -- THAI Gap-PCR primer pair THAI-F and THAI-R for genotype:
[0081] THAI-F: 5'-AAGCGAGAGGAATCACATTC-3' (SEQ ID NO: 5);
[0082] THAI-R: 5'-CTTGGATCTGCACCTCTG-3' (SEQ ID NO: 6);
[0083] (4) The fluorescent probe A1-Prob that sp...
Embodiment 2
[0115] Example 2: The detection effect of the present invention in random genotype samples.
[0116] 1. The composition of the kit:
[0117] With embodiment 1.
[0118] 2. Implementation method:
[0119] With embodiment 1.
[0120] 3. Sample source:
[0121] The samples were 58 random samples (provided by Jinan Xinyue Biotechnology Co., Ltd.), diluted with double distilled water to 20-200ng, as samples to be tested), and 3 normal samples verified by Gap-PCR.
[0122] 4. Data analysis and result judgment:
[0123] Calculate according to the data analysis formula of the present invention, and determine the relative copy numbers of A1 and A2 obtained by detecting according to the result according to the judgment standard of the aforementioned Table 1, the results are as shown in the following Table 5 and Table 6; when using the present invention While the above-mentioned samples were detected by the kit and method, the existing Gap-PCR method was used for verification, and t...
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