A screening kit for paroxysmal supraventricular tachycardia

A tachycardia and kit technology, applied in fermentation, animal/human peptide, microorganism measurement/testing, etc., can solve the problems of no common disease gene, report, no molecular diagnostic method or kit, etc., and achieve application promising effect

Active Publication Date: 2022-03-11
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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  • A screening kit for paroxysmal supraventricular tachycardia
  • A screening kit for paroxysmal supraventricular tachycardia
  • A 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. Composition of the kit

[0033] 1. Kit composition

[0034] 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.

[0035] 1.1 Amplification Reagents

[0036] 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.

[0037] Table 1 PCR amplification reagents

[0038] components concentration volume PCR mix 2× 600μl Primer pair 10μM 100μl pure water 2ml

[0039] 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.

[0040] Table 2 Primers used for gene amplification

[0041]

[0042] 1.2 ...

experiment example 1

[0072] Experimental example 1 family clinical verification

[0073] 1. Family probands with atrioventricular nodal reentrant tachycardia and their family status

[0074] (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.

[0075] (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

[0080] Experimental example 2 Zebrafish embryo experiment

[0081] 1. Method

[0082] 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, which includes an optional reagent for detecting the c.1700A>G mutation site of human CACNA1B gene; the CACNA1B gene c.1700A >G mutation site refers to the mutation site where the 1700th base in the CACNA1B gene coding region changes from A to G. The kit of the present invention is designed based on the significant correlation between the c.1700A>G mutation of CACNA1B gene and paroxysmal supraventricular tachycardia, and is the first molecular screening kit for paroxysmal supraventricular tachycardia , can assist in the diagnosis of paroxysmal supraventricular tachycardia (especially atrioventricular nodal reentrant tachycardia), contribute to prenatal and postnatal care, and can guide doctors to prescribe the right medicine, with a good application prospect.

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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Patent Type & Authority Patents(China)
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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