Respiratory pathogenic microorganism detection primer combination, kit and application thereof

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

Active Publication Date: 2021-05-28
美格医学检验所(广州)有限公司
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  • 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

Method used

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  • Respiratory pathogenic microorganism detection primer combination, kit and application thereof
  • Respiratory pathogenic microorganism detection primer combination, kit and application thereof
  • Respiratory pathogenic microorganism detection primer combination, kit and application thereof

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0067]Example 1 Detection primer design and amplification method of respiratory pathogen microorganism

[0068]This embodiment is directed to common respiratory pathogen microorganisms: pneumoniae, pneumonia colostrum, parrot hot coat, Mycobacterium tuberculosis, white candida, Yeiri Pneumacterium, human herpes virus 4 (EBV), human herpes virus 5 ( CMV), respiratory syncytial virus (RSV) and nose-specific species-specific primers, primer sequences are shown in Table 1:

[0069]Table 1 Respiratory pathogen microbial detection primer group

[0070]

[0071]

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

[0073](1) TNA (DNA / RNA) of the respiratory sample was obtained, and CDNA was synthesized by RNA under the action of inverse transcriptase.

[0074](2) The first round of amplification is made by DNA and cDNA as a template:

[0075]When the first round of PCR amplification, the above primer sequence i...

Example Embodiment

[0095]Example 2 2 Parts of alveolar lavage liquid samples

[0096]Collection A1, A2 suspected respiratory infection patients with alveolar lavage liquid samples.

[0097]The clinical symptoms of A1 were: intermittent fever for 2 days, the highest body temperature was 39.7 ° C, and the heat was hot. Clinical diagnosis is: 1, sepsis; 2, fungal infection; 3, pneumonia (severe) pathogen. The white blood cell count was 14.74 × 109 / L, HGB: 110g / L; PCT: 306 × 109 / L; CRP: 20.98 mg / L. Other test results: congenital immunodeficiency. Antibiotic: Mei Luo Pusi, Pipellanzoliazazama. According to clinical symptoms and diagnosis, it is judged that it may have a respiratory pathogen microorganism infection.

[0098]The clinical symptoms of patients A2 are: cough, cough, fever, clinical diagnosis as: suspected pneumonia, pneumonia infection. Antibiotic is used as Ampera. According to clinical symptoms and diagnosis, it is judged that it may have a respiratory pathogen microorganism infection.

[0099]Wi...

Example Embodiment

[0159]Example 3 2 Samples of sputum samples of microbial detection

[0160]Two suspected sputum samples B1, B2 were collected.

[0161]B1 patient clinical symptoms: diabetes, lung shadow. Clinical diagnosis: lung CT found suspicious tuberculosis in lungs and mediastinats. Blood routine: WBC: 7.3 × 109 / L; LY%: 21%; NEU%: 6%. According to clinical symptoms, it is determined that it may have pathogenic microbial infections, and there may be a bacterial infection.

[0162]B2 patients clinical symptoms are: fever, difficulty breathing. Clinical diagnosis: 1, severe pneumonia; 2, pulmonary hemorrhage; 3, respiratory failure; 4, respiratory syncytial viral pneumonia; 5, stem cell transplantation; 6, viscous polysaccharide (IV). Blood routine: WBC: 0.7 × 109 / L: ly%: 1%: NEU%: 92%, CRP: 2.23. According to clinical symptoms, it is determined that there may be pathogenic microorganisms, which may be a respiratory virus infection.

[0163]With the primer combination and method of Example 1, the sputum sam...

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Abstract

The invention discloses a respiratory tract pathogenic microorganism detection primer combination, a kit and a method, and belongs to the technical field of microorganism detection. The respiratory tract pathogenic microorganism detection primer combination comprises primers with nucleotide sequences as shown in SEQ ID No.1 to SEQ ID No.20 respectively. According to the method, aiming at a clinical suspected respiratory tract infection sample, a multiplex PCR technology is combined with a targeted sequencing technology to analyze common pathogenic microorganisms in the sample, and the method can be used for detecting pathogenic microorganisms such as bacteria, fungi, viruses (DNA and RNA viruses), mycoplasmas and chlamydia, is simple, rapid and accurate, and is beneficial to clinical precise 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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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升詹太平蒋华
Owner 美格医学检验所(广州)有限公司
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