Bronchoalveolar lavage fluid collection method for 16S rRNA gene detection and detection method

A technology of alveolar lavage fluid and gene detection, applied in the field of biomedicine, to achieve the effect of less pollution and good repeatability

Inactive Publication Date: 2015-04-01
张清玲 +3
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Problems solved by technology

In view of the importance of 16S rRNA gene sequence analysis technology, bronchoalveolar lavage fluid is the most accurate specimen for detecting lower respiratory tract flora and the least oral contamination, and there is currently no bronchoalveolar lavage fluid for 16S rRNA gene detection in my country. It is necessary to provide an effective and reliable method for collecting alveolar lavage fluid for the detection of respiratory bacterial 16S rRNA gene sequence

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  • Bronchoalveolar lavage fluid collection method for 16S rRNA gene detection and detection method

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Embodiment 1

[0054] 1. Case collection

[0055] The research object was 1 case of the test subject.

[0056] 2. Main instruments and reagents:

[0057] Sodium chloride; normal saline (Dongguan Puji Pharmaceutical Co., Ltd.), HE staining solution, sputum genomic DNA extraction kit, ultrasonic nebulizer, low-temperature refrigerator; TGL-16G centrifuge (Shanghai Anting Scientific Instrument Factory), Electronic balance (German Sartorius company); precision pipette (French GILSON company); microscope, lung function instrument, automatic cell smear instrument, 2.0 fluorometer, agarose, electrophoresis apparatus, fiberoptic bronchoscope (Olympus BF 10).

[0058] 3. Collection, preservation and detection of alveolar lavage fluid

[0059] For the above research subjects, the alveolar lavage fluid was collected according to the following steps.

[0060] Step 1: Collect bronchoalveolar lavage fluid;

[0061] Sub-step 11: The subject inhales the bronchodilator by nebulization, stays in the int...

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Abstract

The invention discloses a bronchoalveolar lavage fluid collection method for 16S rRNA gene detection, which comprises the following steps: letting a subject inhale a bronchodilator through atomization, and leaving a venous indwelling needle; continuously supplying oxygen to a nasal catheter in the whole process of collecting bronchoalveolar lavage fluid while monitoring the transcutaneous oxygen saturation and judging the vital signs of the subject according to the transcutaneous oxygen saturation; performing surface anesthesia of superior nasal meatus and pharynx and larynx with 2% lidocaine, and then performing intravenous injection with 2-10mg of midazolam and 125-500mg of alfentanil; and letting a fiber bronchoscope enter the glottis area through the nasal meatus, and anesthetizing the glottis area with 4% lidocaine. The invention aims at providing the bronchoalveolar lavage fluid collection method for 16S rRNA gene detection and the detection method thereof, wherein the collection method is safe, simple and convenient, brings little pollution and has good repeatability.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for collecting and detecting bronchoalveolar lavage fluid for 16S rRNA gene detection. Background technique [0002] The study of respiratory tract microbiome is a relatively new field. 16S rRNA gene sequence analysis is a molecular detection technology commonly used to identify bacterial species, and has been widely used in the study of environmental and clinical specimens. The study of respiratory tract microbiome is a A relatively new field. The 16S rRNA gene is the DNA sequence corresponding to the coding rRNA on the bacterial chromosome, which exists in the chromosomal genome of all bacteria. The 16S rRNA sequence information obtained by needle hybridization is compared with the sequence data in the 16S rRNA database or other data to determine its position in the evolutionary tree, thereby identifying the possible microbial species in the sample. Therefore, compared wit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/24
Inventor 张清玲李乃健邱日皇李时悦
Owner 张清玲
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