Primer and method for simultaneously detecting gene polymorphisms of IL28B and ITPA (inosineadenosine triphatase)

A gene polymorphism, IL28B9917 technology, applied in the field of primers for simultaneous detection of IL28B and ITPA gene polymorphisms, can solve the problems of complex result judgment, large error in test results, low repeatability, etc., and achieve great social significance and accuracy High, improve the effect of the reaction

Inactive Publication Date: 2016-04-20
GUANGZHOU KINGMED DIAGNOSTICS GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Although the current detection method can be used to type IL28B and ITPA, the above method has the defects of large error in detection results, low repeatability, and complicated result judgment

Method used

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  • Primer and method for simultaneously detecting gene polymorphisms of IL28B and ITPA (inosineadenosine triphatase)
  • Primer and method for simultaneously detecting gene polymorphisms of IL28B and ITPA (inosineadenosine triphatase)
  • Primer and method for simultaneously detecting gene polymorphisms of IL28B and ITPA (inosineadenosine triphatase)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment one, primer

[0028] The IL28B and ITPA gene primers provided by the present invention are shown in Table 1, including PCR amplification primers and SNaPshotPCR primers, and the PCR amplification primers and SNaPshotPCR primers are corresponding. All primer sequences provided by the present invention are compared through UCSC database, and there is no known SNP site.

[0029] Table 1 Primers provided by the present invention

[0030]

Embodiment 2

[0031] Embodiment two, the specificity of primer

[0032] The primers provided by the present invention were blasted in UCSC, and the results were as follows:

[0033] 1) IL28B and ITPA gene-specific primers were used for Blast in UCSC. The amplified fragments of all primers covered the corresponding detection sites, and there were no other homologous genes. The amplified fragment of IL28B9917 was located at chr19:39743077+39743211, with a length of 135 bp. Amplified sequence diagram as figure 1; The amplified fragment of IL28B9860 is located at chr19:39738744+39738889, with a length of 146bp. The amplified sequence diagram is as follows figure 2 ; The ITPA amplified fragment is located at chr20:3193710+3193935, the length is 226bp, the amplified sequence diagram is as follows image 3 .

[0034] 2) Use the SNaPshotPCR primers in Table 1, SNaPshot method for detection, the results are as follows Figure 4 As shown, the relative position of each product peak and the bases ...

Embodiment 3

[0035] Example 3, Detection of IL28B and ITPA Gene Polymorphisms

[0036] 1) Extract DNA samples from EDTA anticoagulated peripheral blood. The extraction method refers to the instructions of TIANampBloodDNAKit (purchased from Tiagen, product number DP318), and the DNA samples are diluted to 100ng / μL for later use.

[0037] 2) Prepare the primer mixture: mix the primer concentration ratio of IL28B9917, IL28B9860 and ITPA in the PCR amplification primers at a ratio of 1:1:1, shake and mix well; after short centrifugation, use Q5 for PCR amplification ? Hot start ultra-fidelity 2XMasterMix (purchased from NEB Company, product number M0494L), the reaction system is shown in Table 2, shake and mix well, after a short centrifugation, aliquot 18.0 μL into the labeled PCR reaction tube; pour the labeled PCR reaction tube Add 5.0 μL of primer mixture to the tube, and carry out PCR amplification according to the following procedures: stage 1: 98°C for 3 min; stage 2: 98°C for 10 s; sta...

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Abstract

The invention belongs to the technical field of biological detection, in particular relates to a primer and a method for simultaneously detecting gene polymorphisms of IL28B and ITPA (inosineadenosine triphatase). The primer for simultaneously detecting gene polymorphisms of IL28B and ITPA, provided by the invention, comprises a PCR (polymerase chain reaction) amplification primer and SNaPshot PCR primer. The primer for simultaneously detecting gene polymorphisms of IL28B and ITPA, provided by the invention, has the advantages of high specificity, high detection sensitivity and high accuracy, and detection on the gene polymorphisms of IL28B and ITPA is realized; the primer can be used for guiding dosage adjustment of ribavirin so as to increase reaction of antiviral therapy; meanwhile, the primer can also be used for providing a basis for clinically predicting antiviral individualized treatment effect on hepatitis c, and has great social significance.

Description

technical field [0001] The invention belongs to the technical field of biological detection, and in particular relates to a primer and a method for simultaneously detecting IL28B and ITPA gene polymorphisms. Background technique [0002] Hepatitis C virus, referred to as hepatitis C, is a viral hepatitis caused by hepatitis C virus (Hepatitis C virus, HCV) infection, mainly through blood transmission. Since most patients infected with hepatitis C virus have no obvious symptoms in the acute stage of infection, they are often ignored by people. They are usually not noticed until hepatitis C transforms into chronic hepatitis C or liver cirrhosis and other intractable diseases, which are often missed. The best treatment period for hepatitis C. According to the statistics of the World Health Organization, there are about 140-170 million HCV infected people in the world, and the HCV infection rate in my country is about 3.2%, and there are about 38 million HCV infected people, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6858C12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2565/125
Inventor 赵薇薇胡昌明梁耀铭于世辉燕启江郭周萍
Owner GUANGZHOU KINGMED DIAGNOSTICS GRP CO LTD
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