Kit for detecting common deletion form alpha-thalassemia and using method of kit
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A thalassemia and kit technology, applied in the field of medical detection, can solve the problems of long detection time and long section length, etc.
Inactive Publication Date: 2014-08-06
龙驹
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However, due to -α 3.7 and-alpha 4.2 All are produced by the recombination of large segment homologous genes, so the -α carried by different individuals 3.7 or -alpha 4.2 The gene sequence is not consistent, therefore, the current commonly used diagnostic method is to amplify it by spanning breakpoint PCR (Gap-PCR), but because of the long segment length, the detection time is also long
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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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Abstract
The invention discloses a kit for detecting common deletion form alpha-thalassemia and a using method of the kit. The kit comprises a pair of primers capable of performing simultaneous amplification of a signature sequence A1 in an alpha 1 section and a signature sequence A2 in an alpha 2 section in an alpha-globin gene cluster, a pair of primers capable of performing amplification of a --<SEA> genetype in the alpha-globin gene cluster and a pair of primers capable of performing amplification of a --<THAI> genetype in the alpha-globin gene cluster as well as a fluorescence probe for specific detection of the signature sequence A1 in the alpha 1 section, a fluorescence probe for specific detection of the signature sequence A2 in the alpha 2 section, a fluorescence probe for specific detection of a --<SEA> genetype amplicon and a fluorescence probe for specific detection of a --<THAI> genetype amplicon. As for the alpha-thalassemia globin gene deletion detection, the kit provided by the invention has high sensitivity, stability, accuracy and specificity.
Description
technical field [0001] The invention relates to the technical field of medical detection, in particular to a kit for detecting common deletion α-thalassemia and a use method thereof. Background technique [0002] Thalassemia, referred to as thalassemia, is one of the most common monogenic genetic diseases. The common types of thalassemia are α-thalassemia and β-thalassemia, which are caused by abnormal expression of α-protein gene cluster and β-globin gene cluster respectively. . In my country, Guangxi, Guangdong and Hainan are provinces with high incidence of thalassemia, and the incidence of α-thalassemia in Guangxi is 15.5%. In the Chinese population, the common genotype of the deletion α-thalassemia is -- SEA , -α 3.7 and-alpha 4.2 , and the relatively common Thai-type deletion in Guangxi (-- THAI ). [0003] Homologous gene quantitative technology is a method based on gene homology, using common amplification primers to amplify homologous genes, and finally using ...
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