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Primer set for respiratory tract infection pathogen detection, rapid diagnostic kit and detection method

A rapid diagnosis and respiratory technology, applied in the field of molecular biology, can solve the problems of slow development and no mature technology for pathogen detection of multiple respiratory infections, and achieve the effect of reducing costs

Inactive Publication Date: 2018-07-20
博迪泰(厦门)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, in the field of multiple detection, especially in the field of medium-throughput detection, constant temperature nucleic acid amplification technology including RPA technology has been developing slowly
Restricted by the spectral characteristics of fluorescent dyes used for labeling and the number of fluorescent detection channels, the current RPA technology at home and abroad is still not mature in the detection of multiple respiratory tract infection pathogens

Method used

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  • Primer set for respiratory tract infection pathogen detection, rapid diagnostic kit and detection method
  • Primer set for respiratory tract infection pathogen detection, rapid diagnostic kit and detection method
  • Primer set for respiratory tract infection pathogen detection, rapid diagnostic kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: primer and test kit

[0038] 1. Primer design:

[0039] The primers were designed according to the pathogenic gene sequence of respiratory tract infection in the NCBI gene database, referring to the TwistDx instructionmanual and Primer-BLAST primer design principles. The primer length is about 30-35nt. Since there is no primer design software for RPA at present, a large number of primers were designed and synthesized in the previous work of the present invention, and the primers with high sensitivity and good specificity were screened out for use in the present invention (Table 1).

[0040] 2. Primer synthesis:

[0041] According to the primer sequences shown in Table 1, sequence synthesis was carried out. The 5' end of the forward primer was modified by adding Biotin; the 5' end of the reverse primer was modified by adding 60 T bases.

[0042] Table 1

[0043]

[0044]

[0045] 3. Kit

[0046] The kit (nine-in-one) for the rapid diagnosis of mu...

Embodiment 2

[0048] Embodiment 2: detect multiple respiratory infection pathogens

[0049] The present embodiment adopts the kit described in Example 1 to detect multiple respiratory tract infection pathogens, and the method used in the detection is the solid-phase recombinase polymerase constant temperature gene amplification method, and the specific steps are as follows:

[0050] 1. Clinical Sample Preparation

[0051] Collect the remaining samples after routine examinations of the respiratory tract (throat swabs, nasopharyngeal swabs, etc.), and the collection conditions are: positive clinical samples confirmed to have respiratory pathogenic infection after laboratory examinations (culture method or molecular biology examination method), And negative clinical samples confirmed to be free of respiratory pathogen infection after inspection. The remaining samples collected will be deactivated by heating at 95°C for 10 minutes to avoid the possibility of infection during transportation. C...

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PUM

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Abstract

Belonging to the molecular biology field, the invention relates to a primer set for respiratory tract infection pathogen detection, a rapid diagnostic kit and a detection method. The primer set can achieve one-time detection of the following 9 respiratory tract infection pathogens: influenza A virus, influenza B virus, respiratory syncytial virus, parainfluenza virus type I, parainfluenza virus type II and parainfluenza virus type III, adenovirus, mycoplasma pneumonia, legionella pneumonia, chlamydia pneumonia and human rhinovirus. The invention adopts solid phase recombinase-polymerase constant temperature gene amplification method to detect respiratory multiple infection pathogens. The primer set adopted by the invention for respiratory tract infection pathogen detection has strong specificity and high amplification efficiency, can effectively and rapidly detect the 9 pathogens simultaneously, and realizes multiplex detection.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a primer set for detecting respiratory infection pathogens, a rapid diagnostic kit and a detection method. Background technique [0002] The common diseases of acute respiratory infection include acute upper respiratory tract infection, acute trachea-bronchitis, bronchopneumonia, bronchiectasis, etc., and the infected population is more common in children, which is the first cause of death of young children (<5 years old) in the world. About 5 million children died. According to statistics released by the World Health Organization not long ago, among the top ten causes of death in the world, respiratory tract infection ranks third with an incidence rate of 5.9%, second only to coronary heart disease and stroke, and chronic obstructive pulmonary disease ranks fourth (5.4%) deaths are also mostly related to infection. There are many types of pathogens that cause respiratory tr...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/689C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/689C12Q1/701C12Q2521/107C12Q2521/507C12Q2522/101C12Q2563/107Y02A50/30
Inventor 张睿李博安詹尔昌郭永斌
Owner 博迪泰(厦门)生物科技有限公司
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