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Screening kit for paroxysmal supraventricular tachycardia

A tachycardia, kit technology, applied in the direction of fermentation, animal/human peptide, microorganism determination/inspection, etc., can solve the problem of no consistent disease gene, report, no molecular diagnostic method or kit, etc., to achieve application promising results

Active Publication Date: 2020-04-10
SICHUAN PROVINCIAL PEOPLES HOSPITAL +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, people know very little about its genetics, and there is no report on a disease-causing gene, let alone a report on a molecular diagnostic method or kit.

Method used

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  • Screening kit for paroxysmal supraventricular tachycardia
  • Screening kit for paroxysmal supraventricular tachycardia
  • Screening kit for paroxysmal supraventricular tachycardia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 Kit of the present invention (Sanger sequencing kit)

[0032] 1. Kit composition

[0033] The kit of the present invention includes amplification reagents for amplifying CACNA1B gene (its CDS is shown in SEQ ID NO.3, and its amino acid sequence is shown in SEQ ID NO.4), and reagents for Sanger sequencing.

[0034] 1.1 Amplification Reagents

[0035] The PCR amplification reagent is used to amplify a DNA sequence where the SNP site is located, and its composition is shown in Table 1.

[0036] Table 1 PCR amplification reagents

[0037] components concentration volume PCR mix 2× 600μl Primer pair 10μM 100μl pure water 2ml

[0038] The PCR mixture in Table 1 includes Taq enzymes, dNTPs, magnesium ions and other components required for conventional PCR; the primer pair information is shown in Table 2.

[0039] Table 2 Primers used in gene amplification

[0040]

[0041] 1.2 Sequencing reagents

[0042] The rea...

experiment example 1

[0071] Experimental example 1 family clinical verification

[0072] 1. Family probands with atrioventricular nodal reentrant tachycardia and their family status

[0073] (1) Proband: The proband is a middle-aged female who presents with paroxysmal palpitations, which can be terminated spontaneously within tens of minutes to several hours. Her electrocardiogram shows paroxysmal supraventricular tachycardia, and the heart size is reported by echocardiography. And function is normal, was diagnosed as atrioventricular nodal reentrant tachycardia in Sichuan Provincial People's Hospital. The results of electrocardiogram and intracardiac electrophysiological examination were as follows: figure 1 shown.

[0074] (2) Family: When asking about the medical history, it was found that the patient belonged to a family with tachycardia. After inquiring about the history of tachycardia, dynamic electrocardiogram, esophageal pacing examination, and intracardiac electrophysiological examinati...

experiment example 2

[0079] Experimental example 2 Zebrafish embryo experiment

[0080] 1. Method

[0081] In order to evaluate the function of the human CACNA1B point mutation gene, the inventors first cloned the full-length CDS of human CACNA1B and the point mutation CACNA1B (c.1700A>G) gene, constructed it into a plasmid, obtained a recombinant plasmid, and then transcribed in vitro (using mMESSAGE mMACHINE TM T7 Ultra Transcription Kit (Ambion), operated according to the instructions) to prepare the corresponding mRNA. Zebrafish fertilized eggs were collected according to conventional methods, and then wild-type mRNA and point mutant mRNA of CACNA1B were microinjected into zebrafish embryos at 50 ng / μl (injection volume was 1 nl per embryo), and developed to 48hpf (48h after fertilization) The zebrafish embryos were collected (at the same time, uninjected synchronous embryos were collected as a control), the microscopic morphology of the embryos was observed one by one, and the heart rate wa...

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Abstract

The invention provides a screening kit for paroxysmal supraventricular tachycardia. The screening kit comprises an optional reagent for detecting a human CACNA1B gene c.1700 A>G mutation site, whereinthe c.1700A>G mutation site of the CACNA1B gene refers to a mutation site where the 1700th basic group of the coding region of the CACNA1B gene is changed from A to G. The kit provided by the invention is designed based on the significant correlation between the c.1700A>G mutation of the CACNA1B gene and paroxysmal supraventricular tachycardia; the screening kit for paroxysmal supraventricular tachycardia is a first molecular screening kit for paroxysmal supraventricular tachycardia, can assist in diagnosis of paroxysmal supraventricular tachycardia (especially atrioventricular nodal reentrant tachycardia), is beneficial to prenatal and postnatal care, can guide doctors to apply medicines according to symptoms and has good application prospects.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis kits, in particular to a screening kit for paroxysmal supraventricular tachycardia. Background technique [0002] Cardiac arrhythmia is an important group of diseases in cardiovascular diseases, with an incidence rate of about 3-5% in the population, seriously affecting human health. In recent years, the family aggregation tendency of arrhythmia has attracted extensive attention from domestic and foreign scholars and progress has been made in molecular genetics. Paroxysmal supraventricular tachycardia is a common arrhythmia, including atrioventricular nodal reentrant tachycardia (AVNRT) and atrioventricular reentrant tachycardia, and AVNRT is paroxysmal supraventricular tachycardia The most common form of tachycardia, accounting for about 65% of all patients with paroxysmal supraventricular tachycardia in foreign countries, about 40%-50% in China, the prevalence of AVNRT is about 22.5 / 10,000, a...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11A01K67/027C12N15/85C12N15/90C12N15/12
CPCC12Q1/6883A01K67/0278C12N15/8509C12N15/902C07K14/47C12Q2600/156A01K2207/15A01K2217/072A01K2227/40A01K2267/0375C12Q1/6869
Inventor 李小平罗蓉赵庆顺杨正林贺涛刘明江
Owner SICHUAN PROVINCIAL PEOPLES HOSPITAL
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