A kind of detection primer combination, kit and application of respiratory pathogenic microorganisms

A technology of pathogenic microorganisms and detection kits, which is applied in the field of microbial detection, can solve the problems of low identification accuracy, long cycle, and dependence on cultivation, and achieve the effects of low sequencing data, low cost, and high detection sensitivity

Active Publication Date: 2021-11-09
美格医学检验所(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These technologies have played an important role in the daily identification of pathogenic microorganisms, but there are also certain shortcomings. The former needs to rely on cultivation, the cycle is long, and the identification accuracy is low; the latter requires certain prior knowledge of microbial sequences and cannot deal with unknown or mutated pathogenic microorganisms. Wait

Method used

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  • A kind of detection primer combination, kit and application of respiratory pathogenic microorganisms
  • A kind of detection primer combination, kit and application of respiratory pathogenic microorganisms
  • A kind of detection primer combination, kit and application of respiratory pathogenic microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1 Respiratory tract pathogenic microorganism detection primer design and amplification method

[0068] This embodiment is aimed at common respiratory pathogenic microorganisms: Mycoplasma pneumoniae, Chlamydia pneumoniae, Chlamydia psittaci, Mycobacterium tuberculosis, Candida albicans, Pneumocystis jirovecii, human herpesvirus type 4 (EBV), human herpesvirus type 5 ( CMV), respiratory syncytial virus (RSV) and rhinovirus specific target sequence design specific primers, primer sequences are as shown in table 1:

[0069] Table 1 Primer set for detection of respiratory pathogenic microorganisms

[0070]

[0071]

[0072] A kit for detecting common microorganisms in the respiratory tract can be prepared by using the above primer set. The steps to use are as follows:

[0073] (1) Obtain the TNA (DNA / RNA) of the respiratory tract sample, and under the action of reverse transcriptase, synthesize cDNA using RNA as a template.

[0074] (2) Using DNA and cDN...

Embodiment 2

[0095] Example 2 Detection of pathogenic microorganisms in 2 samples of alveolar lavage fluid

[0096] Collect samples of alveolar lavage fluid from patients with A1 and A2 suspected respiratory tract infections.

[0097] The clinical symptoms of patient A1 were: intermittent fever for 2 days, the highest body temperature was 39.7°C, and persistent fever. The clinical diagnosis is: 1. sepsis; 2. fungal infection; 3. pneumonia (severe disease) pathogen. White blood cell count 14.74×109 / L, HGB: 110g / L; PCT: 306×109 / L; CRP: 20.98mg / L. Other test results: congenital immunodeficiency. Antibiotics: meropenem, piperacillin-tazobactam. According to clinical symptoms and diagnosis, it is judged that there may be respiratory pathogenic microbial infection.

[0098] The clinical symptoms of patient A2 are: cough, sputum, and fever, and the clinical diagnosis is: suspected pneumonia and Mycobacterium tuberculosis infection. The antibiotic that has been used is biapenem. According to...

Embodiment 32

[0159] Example 3 Detection of pathogenic microorganisms in 2 sputum samples

[0160] Collect 2 sputum samples B1 and B2 from suspected infected patients.

[0161] The clinical symptoms of B1 patients are: diabetes, lung shadow. The clinical diagnosis was: lung CT found suspicious tuberculosis in the lungs and mediastinal lymph nodes. Blood routine: WBC: 7.3×10 9 / L; LY%: 21%; Neu%: 6%. According to the clinical symptoms, it is judged that there may be pathogenic microorganism infection, which is more likely to be bacterial infection.

[0162] The clinical symptoms of patient B2 are: fever and dyspnea. The clinical diagnosis was: 1. Severe pneumonia; 2. Pulmonary hemorrhage; 3. Respiratory failure; 4. Respiratory syncytial virus pneumonia; 5. Stem cell transplantation status; 6. Mucopolysaccharidosis (type IV). Blood routine: WBC: 0.7×10 9 / L; LY%: 1%; Neu%: 92%; CRP: 2.23. According to clinical symptoms, it is judged that there may be pathogenic microorganism infection,...

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Abstract

The invention discloses a primer combination, a kit and a method for detecting respiratory pathogenic microorganisms, belonging to the technical field of microorganism detection. The primer combination for detecting respiratory pathogenic microorganisms includes primers respectively having the nucleotide sequences shown in SEQ ID Nos. 1-20. The method of the present invention is aimed at clinical suspected respiratory tract infection samples, using multiplex PCR technology combined with targeted sequencing technology to analyze common pathogenic microorganisms in the samples, and can be used for pathogenic microorganisms including bacteria, fungi, viruses (DNA and RNA viruses), mycoplasma, chlamydia, etc. The detection is simple, fast and accurate, which is helpful for accurate clinical diagnosis and treatment.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, and in particular relates to a combination of primers for detection of respiratory pathogenic microorganisms, a kit and applications thereof. Background technique [0002] Common pathogenic microorganisms in the respiratory tract mainly include viruses, bacteria, mycoplasma, chlamydia, and rickettsia. According to statistics, more than 90% of acute respiratory infections are caused by viruses. One pathogenic microorganism can cause multiple clinical symptoms, and the same clinical manifestation can be caused by multiple caused by pathogenic microorganisms. Bacteria are less likely to cause respiratory tract infections than viruses. Once infected, the symptoms are more serious and the localization is more obvious, such as tonsillitis, otitis media, and sinusitis. Viruses include EBV, CMV, HSV, HHV, bocavirus, adenovirus, rhinovirus, coronavirus, respiratory syncytial virus, influe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6869C12Q1/689C12Q1/6893C12Q1/6895C12Q1/70C12N15/11
CPCC12Q1/6869C12Q1/689C12Q1/6893C12Q1/6895C12Q1/701C12Q1/705C12Q2600/16C12Q2531/113C12Q2537/143C12Q2535/122
Inventor 陈澎明杨仁涛龚浩王东生詹太平蒋华
Owner 美格医学检验所(广州)有限公司
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