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Human parainfluenza virus distinguishing and quantitative detection regent kit

A detection kit and the technology of the kit, which are used in the determination/inspection of microorganisms, material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of inability to diagnose parainfluenza virus and limitations of clinical application.

Active Publication Date: 2009-10-07
广州达安临床检验中心有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the culture method generally takes several days to several weeks, therefore, it cannot be used as a method for rapid diagnosis of parainfluenza virus;
[0007] 2) Direct immunofluorescence detection method: It is necessary to collect serum from patients in the acute phase and 10 to 14 days after the illness in the convalescent phase. Only when the antibody in the convalescent phase increases by 4 times can it be of diagnostic value. Therefore, it can only provide a reference for the epidemiology of the virus
These disadvantages limit its clinical application

Method used

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  • Human parainfluenza virus distinguishing and quantitative detection regent kit
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  • Human parainfluenza virus distinguishing and quantitative detection regent kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Embodiment 1. The composition of human parainfluenza virus RT-PCR one-step typing, quantitative detection kit

[0060] 1.1 Extraction reagent: DEPC.H 2 O; RNA extraction solution A (for DEPC.H 2 O diluted, and autoclaved after 10% SiO 2 );

[0061] Trizol; Chloroform.

[0062] 1.2 RT-PCR reaction reagents

[0063] Including HPIV1 reaction tube, HPIV2 reaction tube, HPIV3 reaction tube

[0064] 1.3 Quality control: positive quality control-P (HPIV1, HPIV2 and HPIV3 in vitro transcription RNA mixture); negative quality control-N (normal saline); quantitative reference-S1~S4 (10 4 copies / ml~10 7 copies / ml HPIV1 or HPIV2 or HPIV3 in vitro transcribed RNA)

Embodiment 2

[0065] Embodiment 2. Use human parainfluenza virus RT-PCR one-step typing, quantitative detection kit to detect the infection of human parainfluenza virus in the sample to be tested

[0066] 2.1 Collection, preservation and transportation of specimens: Detectable specimens include sputum, throat swab and bronchoalveolar lavage fluid. The collection methods are as follows: ① Sputum: suck sputum under negative pressure, seal it and send it for inspection; ③Bronchoalveolar lavage fluid: About 1ml of bronchoalveolar lavage fluid was collected by clinicians, sealed and sent for examination. Specimens can be used for testing immediately, or can be stored at -70°C for testing, and the storage period is 6 months. Specimens were transported using 0°C ice cubes.

[0067]2.2 Nucleic acid extraction: ①Pipe 100μl of sputum sample into a 1.5ml sterilized centrifuge tube; centrifuge throat swab sample liquid or bronchoalveolar lavage fluid at 12,000rpm for 5 minutes, discard the supernatan...

Embodiment 3

[0077] Embodiment 3. The composition of human parainfluenza virus RT-PCR two-step method typing, quantitative detection kit

[0078] 3.1 Extraction reagent: same as 2.1 in Example 2

[0079] 3.2 RT-PCR Reagents

[0080] Including mixed reverse transcription reaction tube (containing HPIV1, HPIV2, HPIV3 primers), reverse transcriptase system, HPIV1 reaction tube, HPIV2 reaction tube, HPIV3 reaction tube

[0081] 3.3 quality control product: the same as 2.3 in Example 2

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Abstract

The invention relates to three reagent kits for detecting human parainfluenza viruses 1, 2, 3 type real-time fluorescence polymerase chain reaction, which respectively adopts a one-step method RT-PCR, a two-step method RT-PCR and a multicolor fluorescence method RT-PCR to distinguish types of the human parainfluenza viruses of multi-type specimens and fix an amount of the human parainfluenza viruses of the multi-type specimens. The detection methods of the reagent kits have simple operation, short consuming time and high sensitivity and specificity and can be extensively used in a plurality of fields, such as the auxiliary diagnosis of the infection of the human parainfluenza viruses, clinical medicine direction, epidemiology retrospective study, and the like.

Description

technical field [0001] The invention relates to a real-time fluorescent polymerase chain reaction detection kit for three types of human parainfluenza virus types 1, 2 and 3. The three kits use one-step RT-PCR, two-step RT-PCR and multi-color fluorescent RT-PCR reactions to type and quantify human parainfluenza virus in various types of specimens, and can be widely used for human parainfluenza virus Auxiliary diagnosis of infection, guidance of clinical medication and epidemiological retrospective research and other fields. Background technique [0002] Human parainfluenza virus (HPIV) is a single cell RNA virus belonging to the paramyxovirus family. According to its antigenicity and heredity, it can be divided into four serotypes, that is, human parainfluenza virus type 1 (HPIV1). , human parainfluenza virus type 2 (HPIV2), human parainfluenza virus type 3 (HPIV3) and human parainfluenza virus type 4 (HPIV4). Human parainfluenza virus is a common pathogen of acute viral r...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
Inventor 陈颖王方金周高英李明程钢何蕴韶
Owner 广州达安临床检验中心有限公司
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