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A detection kit for human parainfluenza virus nucleic acid extraction-free gene typing

A human parainfluenza virus and detection kit technology, applied in the fields of biotechnology and medicine, can solve the problems of long time-consuming and low detection efficiency of single-plex detection, and achieve the effects of good accuracy, high precision and reliability.

Inactive Publication Date: 2018-10-19
WUXI ENTRY EXIT INSPECTION & QUARANTINE BUREAU PEOPLES REPUBLIC OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to solve the defects of direct amplification of samples, long time-consuming single detection and low detection efficiency in the prior art, and provide a rapid genotyping detection kit for human parainfluenza virus nucleic acid extraction. above question

Method used

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  • A detection kit for human parainfluenza virus nucleic acid extraction-free gene typing
  • A detection kit for human parainfluenza virus nucleic acid extraction-free gene typing
  • A detection kit for human parainfluenza virus nucleic acid extraction-free gene typing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0133] 1. Sample preparation in the sample processing area

[0134] 1.1. This kit does not require nucleic acid extraction, and can directly amplify and detect viral nucleic acid in throat swab samples and serum samples. Before the experiment, the throat swab samples only need to be thawed and vortexed, and blood samples need to be centrifuged to obtain serum;

[0135] 2. Prepare PCR reagents in the reagent preparation area

[0136] 2.1. Prepare the PCR reaction solution according to the following composition (n is the number of reaction tubes):

[0137] HPIV PCR reaction buffer 1 is 19 μl×n, HPIV1 / 2 / 3 / 4 probe mixture is 2 μl×n, enzyme system 3 is 2 μl×n, and the PCR reaction buffer and primer-probe mixture are fully prepared before use. To dissolve, the enzyme system needs to be centrifuged before use to ensure that all enzymes are concentrated at the bottom;

[0138] 2.2. Dispense the PCR reaction solution into PCR reaction tubes according to 23 μl / tube, and move the react...

Embodiment 2

[0157] Three batches of human parainfluenza virus RNA detection kits were used to carry out 10 repeated detections of the positive control, negative control, medium and low concentrations of self-made standard products, and the CV range of the imprecision Ct value within the batch was : HPIV-1 is 0.68~1.42%, HPIV-2 is 0.72~1.55%, HPIV-3 is 0.77~1.47%, HPIV-4 is 0.89~1.65%; the CV range of the imprecision Ct value between batches is: HPIV -1 is 1.12-1.19%, HPIV-2 is 1.04-1.40%, HPIV-3 is 1.03-1.52%, HPIV-4 is 0.95-1.40%; the CV values ​​of intra-assay and inter-assay precision Ct values ​​are less than 5 %, the negative controls were all negative, indicating that the kit has good precision, and the test results are as follows in Table 1:

[0158] Table 1

[0159]

Embodiment 3

[0161] Select other pathogens that have homology to the nucleic acid sequence of human parainfluenza virus, are likely to cause the same or similar clinical symptoms, and are normally parasitized or easily complicated at the sampling site, such as influenza A virus, influenza B virus, influenza C virus, nasal Viruses, respiratory syncytial virus, adenovirus, measles virus, rubella virus, and mumps virus are the samples to be tested, and the qualified human parainfluenza virus nucleic acid typing detection kit is used for detection, and the operation is strictly in accordance with the kit instructions The test was carried out on an ABI7500 real-time fluorescent quantitative PCR instrument, and the test results were all negative, indicating that the specificity of the kit was good.

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Abstract

The invention discloses a human parainfluenza virus nucleic acid extraction-free and rapid gene typing detection kit. The kit includes a PCR amplification reagent and a contrast reagent, the PCR amplification reagent is composed of a PCR reaction buffer solution A, an enzyme system B, and a HPIV1 / 2 / 3 / 4 primer probe mixed liquor, and the contrast reagent is mainly composed of a negative contrast and a positive contrast. The kit has advantages of direct sample amplification, fast multiple detection velocity, high sensitivity, good specificity and the like.

Description

technical field [0001] The invention relates to the fields of biotechnology and medicine, in particular to a human parainfluenza virus nucleic acid typing detection kit. Background technique [0002] Human parainfluenza virus (HPIV) belongs to the Paramyxoviridae family and is a negative single-stranded RNA virus. It is a common pathogen of community-acquired respiratory infections. According to the genetic and serological characteristics, HPIV can be divided into type 1, type 2, type 3 and type 4. [0003] Parainfluenza virus infection can occur throughout the year (especially type 3), and in autumn, croup or laryngotracheobronchitis (type 1, 2) can be prevalent in children, usually milder upper respiratory tract infection than reinfection Disease is more common. However, the same type of parainfluenza virus, especially types 1 and 3, can cause multiple infections. Type 1 and Type 3 parainfluenza virus infections are often prevalent in nurseries, primary schools and othe...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/686C12Q1/701C12Q2563/107
Inventor 耿合员汪圣强吴海磊易海华陆正清钱学铭于九洋孙悦邢琛秋雯丁均曹晓蕴
Owner WUXI ENTRY EXIT INSPECTION & QUARANTINE BUREAU PEOPLES REPUBLIC OF CHINA
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